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½þQnôÛY°ºùðt
 %'[	
f]òñ J OÚrå¯‡	H	�
7»È]Œƒ“¬–+ Ê —ÿïa
„šCé		s	}	Ý	æ	]Z)JÙ¨ÕØã­ÊøûŒBú­ü[‡Š��“Ë F¶ÈIÊCN]	\
ü
jPrÙ*lß$¢§ë  B" œš‹ò±Jît›ß-	ƒ	�	c
‘
å
Ÿf>V|Ž¿ÿ˜»Ð  ƒ³#˜og”·Ûzú¶	,
†
«
ÕŒ±AÐàE^ MÁçèóö®ÄÇh(	h	š	É	ú	(
{~½áÖbŒ»ø£ÄÇ X <o8Ñ9Û � ±•¡ÉÉ°¦ñ Xb‰÷â_O	uVÛáq® G ¬0 @SU	ÊËEG L Ü ;K`f×ù1ò•	ª;lˆ6uçû
 .›Þñª	¥¼V³= e k x � ­ Â ,gÜB°à9Ñ6HSc�³Ä$°…%•å o t ÕÔMl‰q¹`
L£û�GµiY¹Ä�Ë1 | Í \Íïdt¨	¤>
„öìI\ûd¬è’� Q 8•“ZìY	
Õ
9ÛCx¹4÷Ü!ª › 9Æ7’¬ì	+	vÔ
PªmoÏï b © îJ	�	À	J
˜
å_ñqR_HZpðh! ‰ $Ù+$âü� 	Â	õ
ˆ<p\ŽçSDÌBUòüÏëfo~Ÿ · ~™Éú?Eu®¶o	è	À
ÅˆÁÖúÍ±\Á	,]° “ }ƒzM¿x„¼	ß	š
`m„§zrj. D _ œŒ¡¨	3	L	v	€	§	4
¦0hÈSÁLÖrÆª­ÁIs|Œ#Ž‘”Íd©X™&|Í, P FXê�½Ê-¼)@	:
L
+Fp®"3�çÈÝ+|ašž¢êùtæLsÁQ Ô AÒ~’3`y	òÛá3eK–¿.¢à ú l¤\B�µÙ%p
Ìk	Îv4þ6÷1" € %®	ƒ†lèeYmÇó\‹×ÚX‚¯úA¬ïûs‰Œ�’•Ê!  µ™ñ¶ÉÚBgsÞŒ	+
…
z}ÔOU‹k¦þaÏ÷] W Wˆöá;Ÿ?R7”ÈD¯ d s w ¬ ¯ešßk5GR�ðFhU�æ Ì î§ë—
Bõ3[c Îî ^ ’ *ùL¾tn	u		Ÿ	ô
/_R�ÀælÅH‹cÎê+˜¯ Ó Ø
,X{ÆàøŠårÀJ¾¡ß5  V r «  Võíd™r‹	ET2ýTÇ Íå ‘ Ò ôìc½qm	.Qzèé4Ìä  ,Ä  qGACNNNNNNGTC,CCAGGCCTGG,GATCCGGATC,CCWGGTACCWGG,CCANNNNNTGG,CCAGGNNNTGG,CCANNNCCTGG,CCAGGNCCTGG,GCACNNNAACGTTNNNGTGC,CACNNNGTG,CCNNNNNNNGG,AACNAACRYGTGCGTGC,GCACGCACRYGTTNGTT,GCACNAACRYGTTNGTGC,AACGCACRYGTGCGTT,GACNNNNNGTC,GAANNNNNNNTTGG,CCAANNNNNNNTTC,CACNNNNGTG,AACACNNNNGTGC,GCACNNNNGTGTT,GCANNNNNNTGC,GAACNNNNNNTCC,GGANNNNNNGTTC,CAGNNNCTG,CCAGGNNCTG,CAGNNCCTGG,CCWGGGCCC,GGGCCCWGG,CCWGGGCCCWGG,AACNNNNNNGTGCACNNNNNNGTT,GACNNNGTC,GAANNNNTTC,CCWGGTACC,GGTACCWGG,CCWGGWCCWGG,ACNNNNGTAYC,GRTACNNNNGT,CCWGGYRCCWGG,GAAGNNNNNNTAC,GTANNNNNNCTTC,AACGGCNNNGTGC,GCACNNNGCCGTT,GCACGGCNNNGTT,AACNNNGCCGTGC,CGANNNNNNTGC,GCANNNNNNTCG,CGATCNNNNTGC,GCANNNNGATCG,TGANNNNNNTCA,TGATCNNNNTCA,TGANNNNGATCA,TGATCNNGATCA,AACCTGCNNGTGC,GCACNNGCAGGTT,GCACCTGCNNGTT,AACNNGCAGGTGC,GCCNNNNNGGC,AACNNNNNNGTGCMC,GKGCACNNNNNNGTT,CCWGGNCCWGG,AACTGGGNNGTGC,GCACNNCCCAGTT,GCACTGGGNNGTT,AACNNCCCAGTGC,GACNNNNGTC,GAGNNNNNCTC,CCWGGTCTC,GAGACCWGG,GATNNNNATC,GATCNNNATC,GATNNNGATC,GATCNNGATC,CCWGGCGCCWGG,ACNNNNNCTCC,GGAGNNNNNGT,AACNNNNNNGTGCAG,CTGCACNNNNNNGTT,GATCATGATC,CCWGGNNCCWGG,AACTGGNNNGTGC,GCACNNNCCAGTT,GCACTGGNNNGTT,AACNNNCCAGTGC,AACNAACCGGTGCGTGC,GCACGCACCGGTTNGTT,GCACNAACCGGTTNGTGC,AACGCACCGGTGCGTT,GCANNNNNTGC,GCACNNNNNNGTTCGAACNNNNNNGTGC,CCTNNNNNAGG,GCNNNNNNNGC,CCANNNNNNTGG,CCAGGNNCCTGG,CCTGGCCAGG,CAANNNNNGTGG,CCACNNNNNTTG,TTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAA,GCACNNNNNNGTTTAAACNNNNNNGTGC,CCAGGNCCY,RGGNCCTGG,CCWGGGNCCTGG,CCAGGNCCCWGG,AAGNNNNNCTT,GAYNNNNNVTC,GABNNNNNRTC,GAYNNNNNRTC,GATCNNNNVTC,GABNNNNGATC,GAYNNNNGATC,GATCNNNNRTC,GATCNNNGATC,GTYAACNNNNNNGTGC,GCACNNNNNNGTTRAC,GCACNNNNNNGTTAACNNNNNNGTGC,GATCNGATC,TAACTATAACGGTCCTAAGGTAGCGA,TCGCTACCTTAGGACCGTTATAGTTA,CTCTCTTAAGGTAGC,GCTACCTTAAGAGAG,TAGGGATAACAGGGTAAT,ATTACCCTGTTATCCCTA,GTTAACNNNNNNGTGC,GCACNNNNNNGTTAAC,AACNAACGCGTGCGTGC,GCACGCACGCGTTNGTT,GCACNAACGCGTTNGTGC,AACGCACGCGTGCGTT,TCCAACNNNNNNGTGC,GCACNNNNNNGTTGGA,CAYNNNNRTG,AACAYNNNNGTGC,GCACNNNNRTGTT,GTTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAAC,CCWGGCGCC,GGCGCCWGG,CCWGGNNCC,GGNNCCWGG,GATCGCGATC,AACNAACATGTGCGTGC,GCACGCACATGTTNGTT,GCACNAACATGTTNGTGC,AACGCACATGTGCGTT,TGGCAAACAGCTATTATGGGTATTATGGGT,ACCCATAATACCCATAATAGCTGTTTGCCA,ATCTATGTCGGGTGCGGAGAAAGAGGTAATGAAATGG,CCATTTCATTACCTCTTTCTCCGCACCCGACATAGAT,AACNNGCACGTGCNNGTT,GAACNNNNNCTC,GAGNNNNNGTTC,CCAGGWCCY,RGGWCCTGG,CCWGGGWCCTGG,CCAGGWCCCWGG,CCAGGWCCTGG,TTATAACNNNNNNGTGC,GCACNNNNNNGTTATAA,GCACNNNNNNGTTATAACNNNNNNGTGC,GAACNNNNNNTAC,GTANNNNNNGTTC,GGCCNNNNNGGCC,AACNNACTAGTGC,GCACTAGTNNGTT,AACTAGTNNGTGC,GCACNNACTAGTT,AACNAACTAGTGCGTGC,GCACGCACTAGTTNGTT,GCACNAACTAGTTNGTGC,AACGCACTAGTGCGTT,AACNNNAACGTGCNNGTGC,GCACNNGCACGTTNNNGTT,GCACCCANNNGTT,AACNNNTGGGTGC,TTAACNNNNNNGTGC,GCACNNNNNNGTTAA,CACNNNNNNTCC,GGANNNNNNGTG,CACNNNNGATCC,GGATCNNNNGTG,GATCTAGATC,CCANNNNNNNNNTGG,CCAGGCCT,AGGCCTGG,CCTCGAGG,GATCCGGA,TCCGGATC,GGCGCGCC,GCGATCGC,CCWGGWCC,GGWCCWGG,GATCGATC,GATCATGA,TCATGATC,GCGGCCGC,CCTGGCCR,YGGCCAGG,CCWGGNCC,GGNCCWGG,GGCCGGCC,RTGCGCAY,GATCNNGA,TCNNGATC,CGCGCGCG,CCTGGCCA,TGGCCAGG,CGCCGGCG,GTTTAAAC,GATCGCGA,TCGCGATC,TTAATTAA,VCTCGAGB,CCTGCAGG,CRCCGGYG,CGTCGACG,ATTTAAAT,GCCCGGGC,GATCTAGA,TCTAGATC,CACCTGC,GCAGGTG,CCTNAGG,GATCGAT,ATCGATC,CCTCAGC,GCTGAGG,GCTNAGC,CCTNAGC,GCTNAGG,GCTCTTC,GAAGAGC,GGTNACC,CGGWCCG,ACCWGGT,RGGNCCY,CCTGGAG,CTCCAGG,GGTGATC,GATCACC,GATCNGA,TCNGATC,GGGWCCC,GAAGATC,GATCTTC,CCCWGGG,TCCNGGA,TCCWGGA,RGGWCCY,TTATAA,AGGCCT,GACGTC,GTMKAC,TCCGGA,TGCGCA,ACCTGC,GCAGGT,GGTACC,AGTACT,GGYRCC,CCGCTC,GAGCGG,AACGTT,YGGCCR,RAATTY,CTGAAG,CTTCAG,CACGTG,GRCGYC,AGCGCT,CTTAAG,ACRYGT,ACCGGT,ACTAGT,CACGTC,GACGTG,GWGCWC,GTGCAC,CYCGRG,GGGCCC,ATTAAT,CCTAGG,TTCGAA,GCTAGC,GKGCMC,TGGCCA,GGATCC,GRGCYC,ATCGAT,CACGAG,CTCGTG,GGCGCC,GAAGAC,GTCTTC,GCATGC,GTATCC,GGATAC,TGATCA,ACTGGG,CCCAGT,CTRYAG,RGCGCY,AGATCT,GGNNCC,CTGGAG,CTCCAG,CTTGAG,CTCAAG,CGATCG,GGTCTC,GAGACC,YACGTR,CCNNGG,GAATGC,GCATTC,WCCGGW,GCAATG,CATTGC,RCCGGY,GCGCGC,GAGGAG,CTCCTC,CGGCCG,CCCAGC,GCTGGG,GTGCAG,CTGCAC,CGRYCG,CGTACG,CGTCTC,GAGACG,CCATGG,TCGCGA,GDGCHC,TGTACA,TCATGA,ACATGT,GTATAC,CCWWGG,CTCTTC,GAAGAG,GCNNGC,CCRYGG,GTTAAC,RCATGY,TACGTA,RGATCY,GCGATG,CATCGC,GCAGTG,CACTGC,CCCGGG,CCGCGG,TTTAAA,GGCGGA,TCCGCC,GAGCTC,GATATC,CTGRAG,CTYCAG,CAGCAG,CTGCTG,GAATTC,ATGCAT,CATATG,GTYRAC,AAGCTT,GTNNAC,TCNNGA,CAATTG,ACGCGT,TCCRAC,GTYGGA,GCCGGC,CMGCKG,GCCGAG,CTCGGC,CTCGAG,CTGCAG,CAGCTG,GTCGAC,CTYRAG,AATATT,GATCGA,TCGATC,GACCGA,CACCCA,TCGGTC,TGGGTG,WGTACW,TARCCA,TGGYTA,TCTAGA,GGATC,GATCC,CCWGG,GTCTC,GAGAC,GCWGC,GGNCC,CCSGG,GGTGA,TCACC,GGWCC,GCAGC,GCTGC,CCATC,GATGG,ACGGC,GCCGT,GCNGC,CCNGG,GCATC,GATGC,ACTGG,CCAGT,GGATG,CATCC,CTCAG,CTGAG,GGGAC,GTCCC,ACNGT,CTNAG,CAGTG,CACTG,GACGC,GCGTC,CCCGC,GCGGG,GANTC,CGWCG,TCNGA,CCTTC,GAAGG,GTNAC,GAAGA,TCTTC,GAGTC,GACTC,GTSAC,GAWTC,GCSGC,CASTG,ATGAA,TTCAT,ACGGA,TCCGT,CGCG,CCGC,GCGG,GTAC,AGCT,GCGC,CTAG,GATC,GGCC,CCGG,CATG,RGCY,YATR,ACGT,TGCA,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                                                                                                                                                                                                                                                                                                                                     .                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          	                                                                                                                                                                                                                                                                         -                
                                            	                                            
                      	                                          
         $            	                        ;            
                              	                              	                     	            
         ,   .   (         6            !      d   Q                     C                                                                                                                                                                .                                                                                                                                                                                                                                                                                                          	                                                                                                     -      
                	                
        	              
   $    	        ;    
          	          	       	    
   , . (   6    !  d Q       C   l"  mF  mf  m†  q  u¼  {Vð�	¤4ï  V¤µx�5G¼	p
š
õ
Yòæ4as‘–üêëîš~îsP ¾[	%
 ëò	Fn\»L
pjDØ‰�Ø‰�‰�‰� ýŒÏa  F a  Õ«Ëì(šÛÚñ2jsð
¶
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  r<Notes>
<UUID>3ee2419c-15af-43bb-a972-48b842679ac8</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Description>synthetic circular DNA</Description>
<Created UTC="16:0:0">2017.6.14</Created>
<LastModified UTC="2:17:42">2025.7.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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  %<Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Description>synthetic circular DNA</Description>
<Created>2017.6.15</Created>
<LastModified>2017.6.15</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)."/></Q><Q name="bound_moiety"><V text="lac repressor encoded by lacI"/></Q></Feature><Feature recentID="7" name="T7 prom" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4984-5002" color="#ffffff" type="standard"/><Q name="note"><V text="promoter for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="6" name="lacI prom" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4598-4675" color="#ffffff" type="standard"/><Q name="gene"><V text="lacI"/></Q></Feature><Feature recentID="5" name="lacI" directionality="2" translateFirstCodonAsMet="1" translationMW="38559.52" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="3515-4597" color="#993366" type="standard" translated="1"/><Q name="note"><V text="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)."/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="lacI"/></Q><Q name="product"><V text="lac repressor"/></Q></Feature><Feature recentID="4" name="rop" directionality="2" translateFirstCodonAsMet="1" translationMW="7196.20" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2515-2706" color="#993366" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="rop"/></Q><Q name="product"><V text="Rop protein, which maintains plasmids at low copy number"/></Q></Feature><Feature recentID="3" name="bom" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2271-2413" color="#a6acb3" type="standard"/><Q name="note"><V text="basis of mobility region from pBR322"/></Q></Feature><Feature recentID="2" name="ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1497-2085" color="#ffff00" type="standard"/><Q name="note"><V text="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"/></Q><Q name="direction"><V text="RIGHT"/></Q></Feature><Feature recentID="1" name="KanR" directionality="2" translationMW="30979.95" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="560-1375" color="#993366" type="standard" translated="1"/><Q name="note"><V text="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="aph(3')-Ia"/></Q><Q name="product"><V text="aminoglycoside phosphotransferase"/></Q></Feature><Feature recentID="0" name="f1 ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12-467" color="#ffff00" type="standard"/><Q name="note"><V text="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"/></Q><Q name="direction"><V text="RIGHT"/></Q></Feature></Features>
  Ó<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="T7-ter" sequence="TGCTAGTTATTGCTCAGCGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="5282-5301" boundStrand="1" annealedBases="TGCTAGTTATTGCTCAGCGG" meltingTemperature="56"><Component hybridizedRange="5282-5301" bases="TGCTAGTTATTGCTCAGCGG"/></BindingSite><BindingSite simplified="1" location="5282-5301" boundStrand="1" annealedBases="TGCTAGTTATTGCTCAGCGG" meltingTemperature="56"><Component hybridizedRange="5282-5301" bases="TGCTAGTTATTGCTCAGCGG"/></BindingSite></Primer><Primer recentID="0" name="T7" sequence="TAATACGACTCACTATAGGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="4983-5002" boundStrand="0" annealedBases="TAATACGACTCACTATAGGG" meltingTemperature="49"><Component hybridizedRange="4983-5002" bases="TAATACGACTCACTATAGGG"/></BindingSite><BindingSite simplified="1" location="4983-5002" boundStrand="0" annealedBases="TAATACGACTCACTATAGGG" meltingTemperature="49"><Component hybridizedRange="4983-5002" bases="TAATACGACTCACTATAGGG"/></BindingSite></Primer></Primers>
  %<Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Description>synthetic circular DNA</Description>
<Created>2017.6.15</Created>
<LastModified>2017.6.15</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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‚†1!ªq2X8>(]K�QXÙ^¨ö»|ÒË{`†®Þ†_¦ª@¥�¢hb�Ä°[4|gÐn�o˜�°„ªï­§Epê­U˜&†[á˜YZ4«jìa*®Qika0gf¡bjEVWV@+ÌÝ«üU#ß…h•ÊYª)V³)Z¨¥m;j©}d<VÃU¿¦VÕE‘ñ„Š+Ò ]D½æ• {×-P1UòæÐ1ƒ÷œ…ößåµ*¨Ä#ÉSy[þTÿÆ¤N‡ TðÅÈ1ELT985"L-Î3%÷Ýüó:XÏ9Ð3/ÚÏ      ø  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  g<?xml version="1.0"?><Features nextValidID="16"><Feature recentID="15" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-5369" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="mol_type"><V predef="other DNA"/></Q></Feature><Feature recentID="14" name="T7 term" type="terminator" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="26-73" color="#ffffff" type="standard"/><Q name="note"><V text="transcription terminator for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="13" name="6xHis" directionality="2" translationMW="840.86" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="137" maxFusedRunOn="137"><Segment range="140-157" color="#993366" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="6xHis affinity tag"/></Q></Feature><Feature recentID="12" name="T7 tag" directionality="2" translationMW="1098.30" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="123" maxFusedRunOn="123"><Segment range="207-239" color="#993366" type="standard" translated="1"/><Q name="note"><V text="promotes efficient translation in E. coli"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="leader peptide from bacteriophage T7 gene 10"/></Q></Feature><Feature recentID="11" name="thrombin" directionality="2" translationMW="627.76" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="123" maxFusedRunOn="123"><Segment range="243-260" color="#993366" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="thrombin recognition and cleavage site"/></Q></Feature><Feature recentID="10" name="6xHis" directionality="2" translationMW="840.86" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="123" maxFusedRunOn="123"><Segment range="270-287" color="#993366" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="6xHis affinity tag"/></Q></Feature><Feature recentID="9" name="ribosome binding site" type="RBS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="306-328" color="#a6acb3" type="standard"/><Q name="note"><V text="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"/></Q></Feature><Feature recentID="8" name="laco" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="343-367" color="#31849b" type="standard"/><Q name="note"><V text="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)."/></Q><Q name="bound_moiety"><V text="lac repressor encoded by lacI"/></Q></Feature><Feature recentID="7" name="T7 prom" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="368-386" color="#ffffff" type="standard"/><Q name="note"><V text="promoter for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="6" name="lacI prom" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="695-772" color="#ffffff" type="standard"/><Q name="gene"><V text="lacI"/></Q></Feature><Feature recentID="5" name="lacI" directionality="1" translateFirstCodonAsMet="1" translationMW="38559.52" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="773-1855" color="#993366" type="standard" translated="1"/><Q name="note"><V text="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)."/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="lacI"/></Q><Q name="product"><V text="lac repressor"/></Q></Feature><Feature recentID="4" name="rop" directionality="1" translateFirstCodonAsMet="1" translationMW="7196.20" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2664-2855" color="#993366" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="rop"/></Q><Q name="product"><V text="Rop protein, which maintains plasmids at low copy number"/></Q></Feature><Feature recentID="3" name="bom" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2957-3099" color="#a6acb3" type="standard"/><Q name="note"><V text="basis of mobility region from pBR322"/></Q></Feature><Feature recentID="2" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3285-3873" color="#ffff00" type="standard"/><Q name="note"><V text="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="1" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="3995-4810" color="#993366" type="standard" translated="1"/><Q name="note"><V text="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="aph(3')-Ia"/></Q><Q name="product"><V text="aminoglycoside phosphotransferase"/></Q></Feature><Feature recentID="0" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4903-5358" color="#ffff00" type="standard"/><Q name="note"><V text="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature></Features>
  »<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="T7-ter" sequence="TGCTAGTTATTGCTCAGCGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="67-86" boundStrand="0" annealedBases="TGCTAGTTATTGCTCAGCGG" meltingTemperature="56"><Component hybridizedRange="67-86" bases="TGCTAGTTATTGCTCAGCGG"/></BindingSite><BindingSite simplified="1" location="67-86" boundStrand="0" annealedBases="TGCTAGTTATTGCTCAGCGG" meltingTemperature="56"><Component hybridizedRange="67-86" bases="TGCTAGTTATTGCTCAGCGG"/></BindingSite></Primer><Primer recentID="0" name="T7" sequence="TAATACGACTCACTATAGGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="366-385" boundStrand="1" annealedBases="TAATACGACTCACTATAGGG" meltingTemperature="49"><Component hybridizedRange="366-385" bases="TAATACGACTCACTATAGGG"/></BindingSite><BindingSite simplified="1" location="366-385" boundStrand="1" annealedBases="TAATACGACTCACTATAGGG" meltingTemperature="49"><Component hybridizedRange="366-385" bases="TAATACGACTCACTATAGGG"/></BindingSite></Primer></Primers>
  %<Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Description>synthetic circular DNA</Description>
<Created>2017.6.15</Created>
<LastModified>2017.6.15</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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  #­<?xml version="1.0"?><Features nextValidID="18"><Feature recentID="0" name="Pc promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment name="-35" range="4806-4811" color="#ffffff" type="standard"/><Segment range="4812-4828" color="#ffffff" type="standard"/><Segment name="-10" range="4829-4834" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;intI1&lt;/i&gt; (promoter lies within the coding sequence)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;class 1 integron promoter&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="GmR" directionality="1" translationMW="19414.61" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="5023-5556" color="#ccffcc" type="standard" translated="1"/><Q name="allele"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aacC1&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to gentamycin&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;gentamycin acetyltransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALDLFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRSEIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIREEVMHFDIDPSTAT*"/></Q></Feature><Feature recentID="2" name="source" type="source" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6180" color="#ffffff" type="standard"/><Q name="mol_type"><V predef="other DNA"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="3" name="T7 term" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6155" color="#ffffff" type="standard"/><Q name="note"><V text="transcription terminator for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="4" name="6xHis" directionality="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6044" maxFusedRunOn="6044"><Segment range="6024-6041" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="6xHis affinity tag"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="HHHHHH"/></Q></Feature><Feature recentID="5" name="T7 tag" directionality="1" translationMW="1098.30" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6058" maxFusedRunOn="6058"><Segment range="5942-5974" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="promotes efficient translation in E. coli"/></Q><Q name="product"><V text="leader peptide from bacteriophage T7 gene 10"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MASMTGGQQMG"/></Q></Feature><Feature recentID="6" name="thrombin" directionality="1" translationMW="627.76" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6058" maxFusedRunOn="6058"><Segment range="5921-5938" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="thrombin recognition and cleavage site"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="LVPRGS"/></Q></Feature><Feature recentID="7" name="6xHis" directionality="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6058" maxFusedRunOn="6058"><Segment range="5894-5911" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="6xHis affinity tag"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="HHHHHH"/></Q></Feature><Feature recentID="8" name="ribosome binding site" type="RBS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5853-5875" color="#a6acb3" type="standard"/><Q name="note"><V text="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"/></Q></Feature><Feature recentID="9" name="laco" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5814-5838" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="lac repressor encoded by lacI"/></Q><Q name="note"><V text="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)."/></Q></Feature><Feature recentID="10" name="T7 prom" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5795-5813" color="#ffffff" type="standard"/><Q name="note"><V text="promoter for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="11" name="lacI prom" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4598-4675" color="#ffffff" type="standard"/><Q name="gene"><V text="lacI"/></Q></Feature><Feature recentID="12" name="lacI" directionality="2" translateFirstCodonAsMet="1" translationMW="38559.52" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="3515-4597" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="lacI"/></Q><Q name="note"><V text="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)."/></Q><Q name="product"><V text="lac repressor"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"/></Q></Feature><Feature recentID="13" name="rop" directionality="2" translateFirstCodonAsMet="1" translationMW="7196.20" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2515-2706" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="rop"/></Q><Q name="product"><V text="Rop protein, which maintains plasmids at low copy number"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL*"/></Q></Feature><Feature recentID="14" name="bom" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2271-2413" color="#a6acb3" type="standard"/><Q name="note"><V text="basis of mobility region from pBR322"/></Q></Feature><Feature recentID="15" name="ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1497-2085" color="#ffff00" type="standard"/><Q name="direction"><V text="RIGHT"/></Q><Q name="note"><V text="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"/></Q></Feature><Feature recentID="16" name="KanR" directionality="2" translationMW="30979.95" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="560-1375" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="aph(3')-Ia"/></Q><Q name="note"><V text="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes"/></Q><Q name="product"><V text="aminoglycoside phosphotransferase"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature recentID="17" name="f1 ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12-467" color="#ffff00" type="standard"/><Q name="direction"><V text="RIGHT"/></Q><Q name="note"><V text="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"/></Q></Feature></Features>
   —<?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  Æ<Notes>
<UUID>5942e7a3-4714-46c2-9a52-0c6fbacbf808</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pET-28a-GmR</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>synthetic circular DNA</Description>
<Created UTC="16:0:0">2017.6.14</Created>
<LastModified UTC="8:5:41">2026.3.7</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y      K         
?????                                                                                                       ÿ d      DO –              E              ÿÿÿÿYô? P                                                                                                                                                              ¯<StrandColors><TopStrand><ColorRange range="4776..5586" colors="green"/></TopStrand><BottomStrand><ColorRange range="4776..5586" colors="green"/></BottomStrand></StrandColors>   3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
   )<?xml version="1.0"?><CustomEnzymeSets/>
