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!!$ë$R(Þ(-*.*+ª+«+æ+þ+Ï-'3  =`  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                  
                     .                                                                                                                                                                                                                                                                                                           E                                                         
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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="6" name="SV40 NLS" directionality="1" translationMW="883.15" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="126" maxRunOn="14078" maxFusedRunOn="14078" detectionMode="exactProteinMatch"><Segment range="7011-7031" 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="7" name="superfolder GFP" directionality="1" translationMW="26806.88" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="1409" maxFusedRunOn="1409" detectionMode="exactProteinMatch"><Segment range="693-1406" color="#05fd14" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;GFP variant that folds robustly even when fused to poorly folded proteins (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/16369541&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/16369541&quot;&gt;PÃ©delacq et al., 2006&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"/></Q></Feature><Feature recentID="8" name="T7Te terminator" type="terminator" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1505-1532" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;phage T7 early transcription terminator&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T1 terminator" type="terminator" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1418-1489" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="Shine-Dalgarno sequence" type="RBS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12008-12016" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;full consensus sequence for ribosome-binding sites upstream of start codons in &lt;i&gt;E. coli&lt;/i&gt;; complementary to a region in the 3' end of the 16S rRNA (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pubmed/7528374&quot; title=&quot;https://www.ncbi.nlm.nih.gov/pubmed/7528374&quot;&gt;Chen et al., 1994&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="RBS" type="RBS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="675-686" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;strong bacterial ribosome binding site (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659856&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659856&quot;&gt;Elowitz and Leibler, 2000&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5560-6148" 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="13" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1546-1621" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the &lt;i&gt;Streptococcus pyogenes&lt;/i&gt; CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="Cas9(N)" directionality="1" translationMW="66263.37" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" 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text="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"/></Q></Feature><Feature recentID="15" name="KanR" directionality="2" translationMW="30979.95" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="4520-5335" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V 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  	<?xml version="1.0"?><Primers nextValidID="3"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Kan-R" sequence="ATCGCGAGCCCATTTATACC" description="Kanamycin resistance gene, reverse primer"><BindingSite location="5242-5261" boundStrand="0" annealedBases="ATCGCGAGCCCATTTATACC" meltingTemperature="57"><Component hybridizedRange="5242-5261" bases="ATCGCGAGCCCATTTATACC"/></BindingSite><BindingSite simplified="1" location="5242-5261" boundStrand="0" annealedBases="ATCGCGAGCCCATTTATACC" meltingTemperature="57"><Component hybridizedRange="5242-5261" bases="ATCGCGAGCCCATTTATACC"/></BindingSite></Primer><Primer recentID="1" name="pBR322ori-F" sequence="GGGAAACGCCTGGTATCTTT" description="pBR322 origin, forward primer"><BindingSite location="6048-6067" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="6048-6067" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite location="4438-4451" boundStrand="1" annealedBases="CGCCTGTATCTTT" meltingTemperature="37"><Component bases="GGGAAA"/><Component hybridizedRange="4447-4451" bases="CGCCT"/><Component bases="G"/><Component hybridizedRange="4438-4445" bases="GTATCTTT"/></BindingSite><BindingSite simplified="1" location="6048-6067" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="6048-6067" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite simplified="1" location="4438-4451" boundStrand="1" annealedBases="CGCCTGTATCTTT" meltingTemperature="37"><Component bases="GGGAAA"/><Component hybridizedRange="4447-4451" bases="CGCCT"/><Component bases="G"/><Component hybridizedRange="4438-4445" bases="GTATCTTT"/></BindingSite></Primer><Primer recentID="2" name="pENTR-R" sequence="ATGGCTCATAACACCCCTTG" description="pENTR vectors, reverse primer"><BindingSite location="5326-5345" boundStrand="0" annealedBases="ATGGCTCATAACACCCCTTG" meltingTemperature="57"><Component hybridizedRange="5326-5345" bases="ATGGCTCATAACACCCCTTG"/></BindingSite><BindingSite simplified="1" location="5326-5345" boundStrand="0" annealedBases="ATGGCTCATAACACCCCTTG" meltingTemperature="57"><Component hybridizedRange="5326-5345" bases="ATGGCTCATAACACCCCTTG"/></BindingSite></Primer></Primers>
  ž<Notes>
<UUID>bdc083b0-6577-44b7-b2b9-ef366fd28387</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pyEvolvR (enCas9-PolI5M)</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>Expresses enCas9 fused to PolI5M in S. cerevisiae; contains a BsmbI-flanked GFP cassette for sgRNA cloning..</Description>
<Created UTC="20:56:18">2021.5.12</Created>
<LastModified UTC="8:19:56">2023.10.28</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
<References>
<Reference journalName="ACS Synth Biol. 2020 Jun 16. doi: 10.1021/acssynbio.0c00149." title="Targeted Diversification in the S. cerevisiae Genome with CRISPR-Guided DNA Polymerase I." type="Journal Article" authors="Tou CJ, Schaffer DV, Dueber JE" pubMedID="32485105" journal="ACS Synth Biol. 2020 Jun 16. doi: 10.1021/acssynbio.0c00149."/>
</References>
</Notes>
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   )<?xml version="1.0"?><CustomEnzymeSets/>
