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Ì›ŽÝÞµ–åæõ.ÆËÌ�ÙM;M‰!  aGGGS  bGGSG  cGSGR  dSGRG  eGRGG  fRGGS  gGGSG  hGSGG  iSGGR  jGGRG  kGRGR  lRGRG  mGRGG  nRGGK  oGGKG  pGKGC  qKGCG  5þ  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                                                                                                                                                                                                                                                                                                                                                    ;                                                                                                                                                     	                                                                                                                                                                                                                                                                                                                                                                                                    	                                   	                                                                                                
                                                                                      "                              +          	                                                                                  :                                                                                   ,                                                                                     %         *   '                                 U   B                                                                            !                  +                     ,   <       ;   (      <   D   9      Ç   L               3      ž   %                                                                                                                                                            ;                                                                     	                                                                                                                                                                                    	             	                                    
                                "            +    	                              :                             ,                               %   * '           U B                          !      +       , <   ; (  < D 9  Ç L     3  ž %  �¦  ’<  ’„  ’þ  ˜ø   6  ©bo¼#dé°é™|±7?d›ü6Až,
é‰ùèŽ·ÓYü°ZƒËXœD"[#$$¾« P[ƒÐfú]­h	;	Šê›+Š"	é+“wTD|±¡?hvÐ#h·f»_Û4 åålÈh:	86A·?fr§ª­ÂÉÒâŽE7	:	f	o	„	Í	è	ë	/8ÙÜù»Î—ÛÄïø*„í´_«´GT€‰Û
bdŽ©Ç4 ¸!ô#džf 	3„ï]œ(¾« (œ¾« (¾« (œ(ƒèÐf˜©¯
$$Åio!	?±NÈîž	æëtA�½A’
œa	_æ^ë	c
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ÐÛž[YÅýø"ò�ßúuž 3ë	á	Š
o5Û—#€zãoåË=Þ­oi
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  0à<?xml version="1.0"?><Features nextValidID="19"><Feature recentID="0" name="AAV2 ITR" type="repeat_region" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" prioritize="1" originalSequence="CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT"><Segment range="6525-6654" color="#ffcc99" type="standard"/></Feature><Feature recentID="1" name="AmpR promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7477-7581" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="AmpR" directionality="1" translationMW="31558.16" type="CDS" allowSegmentOverlaps="0" cleavageArrows="7650" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment name="signal sequence" range="7582-7650" color="#ccffcc" type="standard" translated="1"/><Segment range="7651-8442" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Î²-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature><Feature recentID="3" name="Cas9" directionality="1" translationMW="158313.62" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="2" maxRunOn="6278" maxFusedRunOn="6278" detectionMode="exactProteinMatch"><Segment range="1347-5447" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;generates RNA-guided double strand breaks in DNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Cas9 (Csn1) endonuclease from the&amp;nbsp;&lt;i&gt;Streptococcus pyogenes&lt;/i&gt;&amp;nbsp;Type II CRISPR/Cas system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"/></Q></Feature><Feature recentID="4" name="ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8613-9201" color="#ffff00" type="standard"/><Q name="direction"><V text="RIGHT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="f1 ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6740-7195" color="#ffff00" type="standard"/><Q name="direction"><V text="RIGHT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="Kozak sequence" type="regulatory" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1221-1230" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;vertebrate consensus sequence for strong initiation of translation (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pubmed/3313277&quot; title=&quot;https://www.ncbi.nlm.nih.gov/pubmed/3313277&quot;&gt;Kozak, 1987&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="regulatory_class"><V predef="other"/></Q></Feature><Feature recentID="7" name="EBFP2" directionality="1" translationMW="26905.16" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="0" hitsStopCodon="1"><Segment range="5559-5561" color="#3366ff" type="standard" translated="1"/><Segment name="1a" range="5562-5564" color="#3366ff" type="standard" translated="1" translationNumberingStartsFrom="0"/><Segment range="5565-6278" 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;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/17444659&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/17444659&quot;&gt;Ai et al., 2007&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIKVNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLLEFRTAAGITLGMDELYK*"/></Q></Feature><Feature recentID="8" name="CMV enhancer" geneticCode="-1" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="417-702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="nucleoplasmin NLS" directionality="1" translationMW="1739.18" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6278" maxFusedRunOn="6278" detectionMode="exactProteinMatch"><Segment range="5448-5495" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;bipartite nuclear localization signal from nucleoplasmin&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="KRPAATKKAGQAKKKK"/></Q></Feature><Feature recentID="10" name="SV40 NLS" directionality="1" translationMW="883.15" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="126" maxRunOn="6278" maxFusedRunOn="6278" detectionMode="exactProteinMatch"><Segment range="1302-1322" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;nuclear localization signal of SV40 (simian virus 40) large T antigen&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="PKKKRKV"/></Q></Feature><Feature recentID="11" name="hybrid intron" type="intron" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="982-1209" color="noColor" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;hybrid between chicken Î²-actin (CBA) and minute virus of mice (MMV) introns (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/21476867&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/21476867&quot;&gt;Gray et al., 2011&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="bGH poly(A) signal" type="polyA_signal" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6309-6516" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bovine growth hormone polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="U6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1-241" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="AAV2 ITR" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6525-6665" color="#ffe4c4" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;inverted terminal repeat of adeno-associated virus serotype 2&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="rpt_type"><V predef="inverted"/></Q></Feature><Feature recentID="15" name="3xFLAG" directionality="1" translationMW="2730.72" type="CDS" allowSegmentOverlaps="0" cleavageArrows="1295" consecutiveTranslationNumbering="1" maxRunOn="6278" maxFusedRunOn="6278" detectionMode="exactProteinMatch"><Segment range="1230-1295" color="#cc99b2" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;three tandem FLAGÂ® epitope tags, followed by an enterokinase cleavage site&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="DYKDHDGDYKDHDIDYKDDDDK"/></Q></Feature><Feature recentID="16" name="T2A" directionality="1" translationMW="1830.01" type="CDS" allowSegmentOverlaps="0" cleavageArrows="5549" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="139" maxRunOn="6278" maxFusedRunOn="6278" detectionMode="exactProteinMatch"><Segment range="5499-5552" color="#cc99b2" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;2A peptide from&amp;nbsp;&lt;i&gt;Thosea asigna&lt;/i&gt; virus capsid protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="EGRGSLLTCGDVEENPGP"/></Q></Feature><Feature recentID="17" name="MS2 stem loop" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="285-303" color="#99cc00" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;stem loop that binds the bacteriophage MS2 coat protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="MS2 stem loop" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="378-396" color="#99cc00" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;stem loop that binds the bacteriophage MS2 coat protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  &<?xml version="1.0"?><Primers nextValidID="11"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Amp-R" sequence="ATAATACCGCGCCACATAGC" description="Ampicillin resistance gene, reverse primer"><BindingSite location="7799-7818" boundStrand="1" annealedBases="ATAATACCGCGCCACATAGC" meltingTemperature="58"><Component hybridizedRange="7799-7818" bases="ATAATACCGCGCCACATAGC"/></BindingSite><BindingSite simplified="1" location="7799-7818" boundStrand="1" annealedBases="ATAATACCGCGCCACATAGC" meltingTemperature="58"><Component hybridizedRange="7799-7818" bases="ATAATACCGCGCCACATAGC"/></BindingSite></Primer><Primer recentID="1" name="BGH-rev" sequence="TAGAAGGCACAGTCGAGG" description="Bovine growth hormone terminator, reverse primer. Also called BGH reverse"><BindingSite location="6302-6319" boundStrand="1" annealedBases="TAGAAGGCACAGTCGAGG" meltingTemperature="55"><Component hybridizedRange="6302-6319" bases="TAGAAGGCACAGTCGAGG"/></BindingSite><BindingSite simplified="1" location="6302-6319" boundStrand="1" annealedBases="TAGAAGGCACAGTCGAGG" meltingTemperature="55"><Component hybridizedRange="6302-6319" bases="TAGAAGGCACAGTCGAGG"/></BindingSite></Primer><Primer recentID="2" name="EGFP-N" sequence="CGTCGCCGTCCAGCTCGACCAG" description="EGFP, reverse primer"><BindingSite location="4949-4969" boundStrand="1" annealedBases="GCCGTCCAGCTCGACAG" meltingTemperature="42"><Component bases="CGTC"/><Component hybridizedRange="4965-4969" bases="GCCGT"/><Component hybridizedRange="4953-4961" bases="CCAGCTCGA"/><Component bases="C"/><Component hybridizedRange="4949-4951" bases="CAG"/></BindingSite><BindingSite location="5603-5624" boundStrand="1" annealedBases="CGTCGCCGTCCAGCTCGACCAG" meltingTemperature="67"><Component hybridizedRange="5603-5624" bases="CGTCGCCGTCCAGCTCGACCAG"/></BindingSite><BindingSite simplified="1" location="4949-4969" boundStrand="1" annealedBases="GCCGTCCAGCTCGACAG" meltingTemperature="42"><Component bases="CGTC"/><Component hybridizedRange="4965-4969" bases="GCCGT"/><Component hybridizedRange="4953-4961" bases="CCAGCTCGA"/><Component bases="C"/><Component hybridizedRange="4949-4951" bases="CAG"/></BindingSite><BindingSite simplified="1" location="5603-5624" boundStrand="1" annealedBases="CGTCGCCGTCCAGCTCGACCAG" meltingTemperature="67"><Component hybridizedRange="5603-5624" bases="CGTCGCCGTCCAGCTCGACCAG"/></BindingSite></Primer><Primer recentID="3" name="F1ori-F" sequence="GTGGACTCTTGTTCCAAACTGG" description="F1 origin, forward primer"><BindingSite location="7036-7057" boundStrand="0" annealedBases="GTGGACTCTTGTTCCAAACTGG" meltingTemperature="57"><Component hybridizedRange="7036-7057" bases="GTGGACTCTTGTTCCAAACTGG"/></BindingSite><BindingSite simplified="1" location="7036-7057" boundStrand="0" annealedBases="GTGGACTCTTGTTCCAAACTGG" meltingTemperature="57"><Component hybridizedRange="7036-7057" bases="GTGGACTCTTGTTCCAAACTGG"/></BindingSite></Primer><Primer recentID="4" name="F1ori-R" sequence="AGGGAAGAAAGCGAAAGGAG" description="F1 origin, reverse primer"><BindingSite location="6826-6845" boundStrand="1" annealedBases="AGGGAAGAAAGCGAAAGGAG" meltingTemperature="57"><Component hybridizedRange="6826-6845" bases="AGGGAAGAAAGCGAAAGGAG"/></BindingSite><BindingSite simplified="1" location="6826-6845" boundStrand="1" annealedBases="AGGGAAGAAAGCGAAAGGAG" meltingTemperature="57"><Component hybridizedRange="6826-6845" bases="AGGGAAGAAAGCGAAAGGAG"/></BindingSite></Primer><Primer recentID="5" name="hU6-F" sequence="GAGGGCCTATTTCCCATGATT" description="Human U6 promoter, forward primer"><BindingSite location="0-20" boundStrand="0" annealedBases="GAGGGCCTATTTCCCATGATT" meltingTemperature="58"><Component hybridizedRange="0-20" bases="GAGGGCCTATTTCCCATGATT"/></BindingSite><BindingSite simplified="1" location="0-20" boundStrand="0" annealedBases="GAGGGCCTATTTCCCATGATT" meltingTemperature="58"><Component hybridizedRange="0-20" bases="GAGGGCCTATTTCCCATGATT"/></BindingSite></Primer><Primer recentID="6" name="LKO.1 5'" sequence="GACTATCATATGCTTACCGT" description="Human U6 promoter, forward primer"><BindingSite location="171-190" boundStrand="0" annealedBases="GACTATCATATGCTTACCGT" meltingTemperature="51"><Component hybridizedRange="171-190" bases="GACTATCATATGCTTACCGT"/></BindingSite><BindingSite simplified="1" location="171-190" boundStrand="0" annealedBases="GACTATCATATGCTTACCGT" meltingTemperature="51"><Component hybridizedRange="171-190" bases="GACTATCATATGCTTACCGT"/></BindingSite></Primer><Primer recentID="7" name="pBR322ori-F" sequence="GGGAAACGCCTGGTATCTTT" description="pBR322 origin, forward primer"><BindingSite location="9101-9120" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="9101-9120" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite simplified="1" location="9101-9120" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="9101-9120" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite></Primer><Primer recentID="8" name="pBRforEco" sequence="AATAGGCGTATCACGAGGC" description="pBR322 vectors, upsteam of EcoRI site, forward primer"><BindingSite location="7390-7408" boundStrand="1" annealedBases="AATAGGCGTATCACGAGGC" meltingTemperature="57"><Component hybridizedRange="7390-7408" bases="AATAGGCGTATCACGAGGC"/></BindingSite><BindingSite simplified="1" location="7390-7408" boundStrand="1" annealedBases="AATAGGCGTATCACGAGGC" meltingTemperature="57"><Component hybridizedRange="7390-7408" bases="AATAGGCGTATCACGAGGC"/></BindingSite></Primer><Primer recentID="9" name="pGEX 3'" sequence="CCGGGAGCTGCATGTGTCAGAGG" description="pGEX vectors, reverse primer"><BindingSite location="7330-7352" boundStrand="0" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="7330-7352" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite><BindingSite simplified="1" location="7330-7352" boundStrand="0" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="7330-7352" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite></Primer><Primer recentID="10" name="pRS-marker" sequence="CGGCATCAGAGCAGATTGTA" description="pRS vectors, use to sequence yeast selectable marker"><BindingSite location="7211-7230" boundStrand="1" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="7211-7230" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite><BindingSite simplified="1" location="7211-7230" boundStrand="1" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="7211-7230" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite></Primer></Primers>
  <Notes>
<UUID>01e3c57e-df6b-4d8e-b4e7-063813b13268</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pREDIT_Cas9-MS2-BB_BbsI</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>REDIT backbone for wildtype Cas9, pU6-MS2-gRNA-backbone(BbsI)-CBH-SpCas9-T2A-EBFP.</Description>
<Created UTC="21:43:58">2021.5.12</Created>
<LastModified UTC="21:43:58">2021.5.12</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
<References>
<Reference title="Microbial Single-strand Annealing Proteins Enable CRISPR Gene-editing Tools with Improved Knock-in Efficiencies and Reduced Off-target Effects" journalName="Unpublished" journal="Unpublished" type="Journal Article"/>
</References>
</Notes>
  Y     –         
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   )<?xml version="1.0"?><CustomEnzymeSets/>
