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Ÿf>V|Ž¿ÿ¸¾ú±•§!DY”  ƒ³#˜og”·Ûzú¶	,
†
«
ÕŒ±A3Oþ! MÁçèóö®ÄÇh(	h	š	É	ú	(
{~½áÖbÍÉÙ€ƒÎ,MP X <o8¦d
 � ±•¡É/z Xb‰÷â_O	uƒþ¡ G ¬0 @SU	ÊË®Mw[‘ L Ü ;K`f×ù1ò•	ªp„ .›Þñª	¥¼V e k x � ­ Â ,gÜB°à9Ñ6HSc�³Ä$°…º! o t ÕÔMl‰q¹`
L£û�GµiY¹Ä25be¿ì( | Í \Íïdt¨	¤>
„öìIgîPï
 Q 8•“ZìY	
Õ
9ÛCx¹4ê › 9Æ7’¬ì	+	vÔ
P¤íx b © îJ	�	À	J
˜
å_ñqR_! ‰ $Ù+$âü� 	Â	õ
ˆ<p\ŽçSDÌBÀ;DSV•Ñä(§ · ~™Éú?Eu®¶o	è	À
ÅˆÁÖúÍ±Þó2F…	T‡9 “ }ƒzM¿x„¼	ß	š
`m„Ý{1 D _ œŒ¡¨	3	L	v	€	§	4
¦0hÈSÁLÖrÆª­ÁIs|Œ#Ž‘”-nû=€§1¢ÞáV- P FXê�½Ê-¼)@	:
L
+Fp®"3�çÈÝ+|ašž¢êùtæLs&S^� Ô AÒ~’3`y	òÛá3µ´ ú l¤\B�µÙ%p
Ìk	Îv4ÓÌÙb@^l* € %®	ƒ†lèeYmÇó\‹×ÚX‚¯úÄÐÙH„ªÎéõÿ õ4XŽS(  µ™ñ¶ÉÚBgsÞŒ	+
…
z}ÔOU‹k¦þaÌ2ôf½ãùžÔ”¦Í W Wˆöá;Ÿ?R7”­ d s w ¬ ¯ešßk5GR�ðFhUo Ì î§ë—
Bõ3£íw! ^ ’ *ùL¾tn	u		Ÿ	ô
/_R�ÀælÅH‹¿ò mÜEz†8 Ó Ø
,X{ÆàøŠår´Ø
aèþkœ³  V r «  Võíd™r‹	ET2ýT¢¼âìø�¥n ‘ Ò ôìc½qm	.Qz½¾	¡ijy²m  ,Î  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            	               	                                    
         	         
         
   	            
         (   1   "         2            !       b   I                     H                                                                                                                                                                 ,                                                                          
                                                                                                                                                                                                                               
                                                                                                   ,      
                                 	                          !    
    
    6    	     	            
   	   
   
 	    
   ( 1 "   2    !   b I       H   j½  kç  l  l-  o»  to  yûð�	¤4ï   V¤µx�5G¼	p
š
õ
Yòæ4as‘¿ÀÝ€Þw~Tê›îsP ¾[	%
Àò	FŸ\!
pjD^d^d^d^d ýŒÏa  F a  Õ«Ëì(šÛÚñ2jsð
¶
höøÿÈÏX¯uÏ[/:�ËØ.CO\×X Åããõã
ŽQ=]Tê›Œô>!£I‡IF	…¬OãX'…F^d†`f*	9X™”¦ÚÖÛ>Ès/ÑÁ	ARR>oC	‡¦YÈ �á°"7÷]_ ¡’Å ŒäïÁE�¨³u-~4¾±M½°"7H	æ\‘"	N¥‰—"	¥*
Â'äªåu"	Â¥^ßØ.¦ Âïýà¯î°"7Ç/³¢‹Âƒ\	iøårœ ”SƒR Z hÚH›	°æ"7c§ ³ È ð>'>d™�Âá°È"7÷.]ws$áÑJ
å_×òª„ìx­ï V	™*+¼`¾ÄÌVâÿ…�ÔoÑ#ûŽ‡[R¦aûˆ ØoË:CÐöI,{•OJÉ­³ƒ½	 0Ì¦ "	NÂ¥‰ïý^ßØ—ÿ!Œë\C	Ìo
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  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="0:46:55">2019.7.9</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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  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="0:45:48">2019.7.9</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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  »<?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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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. 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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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  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. 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  »<?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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  !½<?xml version="1.0"?><Features nextValidID="17"><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="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="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="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="2" 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="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="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="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="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="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="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="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="8" name="laco" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5003-5027" 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="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="10" name="6xHis" directionality="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5898" maxFusedRunOn="5898"><Segment range="5083-5100" 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="11" name="thrombin" directionality="1" translationMW="627.76" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5898" maxFusedRunOn="5898"><Segment range="5110-5127" 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="12" name="T7 tag" directionality="1" translationMW="1098.30" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5898" maxFusedRunOn="5898"><Segment range="5131-5163" 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="13" name="6xHis" directionality="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5925" maxFusedRunOn="5925"><Segment range="5905-5922" 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="14" name="T7 term" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5989-6036" color="#ffffff" type="standard"/><Q name="note"><V text="transcription terminator for bacteriophage T7 RNA polymerase"/></Q></Feature><Feature recentID="15" name="source" type="source" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6061" 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="16" name="EBFP" directionality="1" translationMW="26900.04" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="0" maxRunOn="5898" maxFusedRunOn="5898"><Segment range="5179-5181" color="#3366ff" type="standard" translated="1"/><Segment name="1a" range="5182-5184" color="#3366ff" type="standard" translated="1" translationNumberingStartsFrom="0"/><Segment range="5185-5895" color="#3366ff" type="standard" translated="1" translationNumberingStartsFrom="2"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;mammalian codon-optimized&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;enhanced blue variant of GFP&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"/></Q></Feature></Features>
  	ë<?xml version="1.0"?><Primers nextValidID="4"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="R1" sequence="tagctcgagtcacttgtacagctcgtccatgccgag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="5871-5904" boundStrand="1" annealedBases="gctcgagtcacttgtacagctcgtccatgccgag" meltingTemperature="71"><Component bases="ta"/><Component hybridizedRange="5871-5904" bases="gctcgagtcacttgtacagctcgtccatgccgag"/></BindingSite><BindingSite simplified="1" location="5871-5904" boundStrand="1" annealedBases="gctcgagtcacttgtacagctcgtccatgccgag" meltingTemperature="71"><Component bases="ta"/><Component hybridizedRange="5871-5904" bases="gctcgagtcacttgtacagctcgtccatgccgag"/></BindingSite></Primer><Primer recentID="1" name="F1" sequence="tagcgaattcatggtgagcaagggcgaggagctg" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="5171-5201" boundStrand="0" annealedBases="cgaattcatggtgagcaagggcgaggagctg" meltingTemperature="69"><Component bases="tag"/><Component hybridizedRange="5171-5201" bases="cgaattcatggtgagcaagggcgaggagctg"/></BindingSite><BindingSite simplified="1" location="5171-5201" boundStrand="0" annealedBases="cgaattcatggtgagcaagggcgaggagctg" meltingTemperature="69"><Component bases="tag"/><Component hybridizedRange="5171-5201" bases="cgaattcatggtgagcaagggcgaggagctg"/></BindingSite></Primer><Primer recentID="2" name="T7-ter" sequence="TGCTAGTTATTGCTCAGCGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="5974-5993" boundStrand="1" annealedBases="TGCTAGTTATTGCTCAGCGG" meltingTemperature="58"><Component hybridizedRange="5974-5993" bases="TGCTAGTTATTGCTCAGCGG"/></BindingSite><BindingSite simplified="1" location="5974-5993" boundStrand="1" annealedBases="TGCTAGTTATTGCTCAGCGG" meltingTemperature="58"><Component hybridizedRange="5974-5993" bases="TGCTAGTTATTGCTCAGCGG"/></BindingSite></Primer><Primer recentID="3" 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>
  Ç<Notes>
<UUID>284b5a4e-2063-49d1-923e-4e12c9867407</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pET28a-EBFP</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>synthetic circular DNA</Description>
<Created UTC="16:0:0">2017.6.14</Created>
<LastModified UTC="2:25:4">2025.8.27</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y      K         
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   )<?xml version="1.0"?><CustomEnzymeSets/>
