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L‚ª¬D“H{(  =]‹¡þh	º	Ï	
/
ï
A‹Xoª±¾h©Ù' û :AÖixðGJ�•žÎ"èòÌ	kW{¥ëOô~“©ÜåCëP€NQTWZ � ï &m¯Ä	&
K1rÅâhOÍ$9N½ %'2	4	Ï{“ÚÑf] J OÚrå¯‡ž�¼«/<Ñ ÷õÈß) Ê —ÿïa
„šCé·	²}çñQZÑˆÎ�¾MILW!>lo E¤ÐÓÖÙÜ  F¶ÈIÊCN	F	•	6
Å
Ê
%ÑÐpˆÞÄæMžàS˜_i‹% œš‹ò±Jî‹+	B
ˆ
èS¡÷×YÚ²Ê�ð3sáT  ƒ³#˜og”·Û­	în* ú‹I %�µ§ MÁçèóö®ÄÇî	5ÜœÜ=nœï�ò�1UJÖAì X <oW¬‚$ � ±aã	Ÿ
’	=ï: Xb‰÷â_Ô:
“
±
ÃéŠ÷a G ¬0 @S É�>? L Ü ;K`f×ù1ò[	
>
_
Pey	¾0D .›Þª
á
`e0Ê e k x � ­ Â ,gÜB°à9Ñ6HSq		z
×'8˜$ù. o t ÕÔMl‰q¹•‰	�	
ÔÀoõ»)Ý|Í-8ó€z | Í \Íïdt¨	õ¹
²zøj`½ÛÊ Q 8•“ZìzÏÍ‘yI­O·ì-¨^ó › 9Æ7’¬ìx
�ŸêH|~Ä8 b © 	¦	b¾4¾YÓeåÆÓ±  ‰ $Ù+$âü��
86iü°ä„Ð[Ç¸@ƒ¶4¯¸Çèg · ~™Éú?EuÃûÁ
!"*ã\49ü5JnA%Rgu¦ù “ }ƒzM¿x	S	Ó	Hø0SÔáxø³/ D _ œw	¨	½
ì
 §Àêô¨¤Ü<Ç5ÀJæ:!5u½çð —¡âoÅ. P FXê�½Ê-¼)ß	!
i
=´®ÀŸºä"–§[y<QŠŸðw�Õ^mèZ“Àçš Ô AÒ~’3`´ífOU§) ú l¤\B�µÙ%ñ¹	ä@ß}Bê’¨G@Mz" € %®	ƒ†lè1	%
�ÙÍá;gÐÿKNÌö#n8DM¼ÒÕØÛÞ$  µ™ñ¶ÉÚBŠ*	”	‡
4ÛçR ŸùîñŠHÃÉÿßrÕ@¦h W Wˆöá;Ÿ?R«� d s w ¬ ¯ešßk5GR~	^
©
dºÜÉ/ Ì î§ë~¶i§7 ^ ’ *ùL¾tÀ
ë
Gâéóh£ÓÆ4Zà9¼ÿ3ftáø Ó Ø
,<xÌï:vTlþYæ(L~  V r «  Võíd™‰“	æÿ¹È¦qÈ. ‘ Ò ôìc½’	åá¢Åî12}-  ,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                                          	                     
               	            	   
                     ,   /   (         9                   g   H                     G                                                                                                                                                                 ,                                                                                                                                                                                                                                                                                                    	                                                                         	                             /      
                               
                             	         6              	       
     	    	 
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   —<?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  q<Notes>
<UUID>15144748-8855-4af9-ac74-a84f9cb9cf34</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Description>synthetic circular DNA</Description>
<Created UTC="16:0:0">2017.6.14</Created>
<LastModified UTC="7:21:26">2021.6.9</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>
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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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ò„ß<óPX#cPFýt¿ð÷ ø÷ÙýÏÔÎØOÅ Äün¿ÛbÃ�ÏM|ßˆ3Âð÷ËÃ$A±±¿1ZYÇ·™ Z!0eÖ¿îEeª*^¥™öM=¶Ûm·M0ø#ÌÃOfM‡‚MX0o±;ËNŒ<ÞÄß}÷ßxFðã—TüTá(8ãß8MeäÙèîWè@*¨áPAy›Á      ø  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  ¨<?xml version="1.0"?><Features nextValidID="16"><Feature recentID="15" name="source" type="source" swappedSegmentNumbering="1" 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" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5297-5344" color="#ffffff" type="standard"/><Q name="note"><V text="transcription terminator for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="13" name="6xHis" directionality="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5233" maxFusedRunOn="5233"><Segment range="5213-5230" 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="1" translationMW="1098.30" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5247" maxFusedRunOn="5247"><Segment range="5131-5163" 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="1" translationMW="627.76" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5247" maxFusedRunOn="5247"><Segment range="5110-5127" 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="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5247" maxFusedRunOn="5247"><Segment range="5083-5100" 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" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5042-5064" 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" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5003-5027" 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="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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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>
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recentID="8" name="ribosome binding site" type="RBS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5926-5948" 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="lacI prom" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5482-5559" color="#ffffff" type="standard"/><Q name="gene"><V text="lacI"/></Q></Feature><Feature recentID="10" name="lacI" directionality="2" translateFirstCodonAsMet="1" translationMW="38558.17" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="4399-5481" 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="11" name="rop" directionality="2" translateFirstCodonAsMet="1" translationMW="7196.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="3399-3590" 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="12" name="bom" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3155-3297" color="#a6acb3" type="standard"/><Q name="note"><V text="basis of mobility region from pBR322"/></Q></Feature><Feature recentID="13" 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="14" 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><Feature recentID="15" name="MCS" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6051-6096" color="#ffff99" type="standard"/></Feature><Feature recentID="16" name="lac operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5887-5911" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-Î²-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5868-5886" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="T7 terminator" type="terminator" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6181-6228" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
   —<?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  Æ<Notes>
<UUID>5f4cf620-d056-40fb-b976-1df73837e0b8</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pET28a-Str</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>synthetic circular DNA</Description>
<Created UTC="16:0:0">2017.6.14</Created>
<LastModified UTC="17:2:48">2025.9.3</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y      K         
?????                                                                                                       ÿ d      HO  –               E              ÿÿÿÿYô? P                                                                                                                                                              3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
   )<?xml version="1.0"?><CustomEnzymeSets/>
