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b
ñ¬ÊhAÏrÃ Q � o­ÞC`ËW
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9„©ààBú±ÈØ,á& g oOÃ�Š
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l
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ä
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nÔ!ÂIV‡”©îNkP÷Rmôgv¦Ù”å œ*š‚¬,V!ôÿëdƒ•æø¸-«I  2 � šï‘ 
•ÃÐíNBæZŽ*è! �=R¸3H³šºQÛ	Ž	*
/\q×=ä>WÎwvOl½ v¢fu­I¤Mÿ�úú§±1”*[ˆ69xÉ- ! V „ 9iûHfŠ°ôBkÿ'aé	Tå	“5	š	·ñ‡‹²(Qš?.‡"½FâíWø)    � l©â?X|×4Ã=	ë	É
ø:Ù?et­°_?¸Vx{~�„hÑŠÆ 9 1Oõ!@@�cE 0 @W;=ÄL	×	þ	=
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¨
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T
„ñ‘Ì*Ô)Òš-
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{
‰x¬#”ð>~ÕG —öˆ¬«Ç	, ¿ IQñÅ×Šèî¦ÌHñH	l	&
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ç
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m‹2Q¼$Šü *`@Ý{Ì
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cn¢m¦aƒ/…9«  Înc	F
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b&»ÔYõ-šcÃ#²S¢ä® € å™Ø8” óU‚åYn‚öˆóf# kŒ8hu‰¡et¬QSPÚL<œãð†±'“‡wN¼5ìÅ \Ð<?ô]ÃÓ<
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›
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Lý  +ž  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                                                                                                                                                                                                                                                        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  -Ú<?xml version="1.0"?><Features nextValidID="17"><Feature recentID="0" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6717-6808" 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="1" name="SmR" directionality="2" translationMW="29333.24" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="5925-6716" 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;aadA&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to spectinomycin and streptomycin&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside adenylyltransferase (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/2993813&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/2993813&quot;&gt;Murphy, 1985&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK*"/></Q></Feature><Feature recentID="2" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5356-5431" 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="3" name="tet operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5282-5300" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;tetracycline repressor TetR&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;tetO&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial operator O&lt;sub&gt;2&lt;/sub&gt; for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="tet operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5307-5325" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;tetracycline repressor TetR&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;tetO&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial operator O&lt;sub&gt;2&lt;/sub&gt; for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="araC" directionality="2" translationMW="33384.84" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="11-889" color="#993366" 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;araC&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;L-arabinose regulatory protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"/></Q></Feature><Feature recentID="6" name="araBAD promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="916-1200" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;araBAD&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter of the L-arabinose operon of &lt;i&gt;E. coli&lt;/i&gt;; the &lt;i&gt;araC&lt;/i&gt; regulatory gene is transcribed in the opposite direction (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/7608087&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/7608087&quot;&gt;Guzman et al., 1995&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="pSC101 ori" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4494-4716" color="#ffff00" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;low-copy replication origin that requires the Rep101 protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="Rep101(Ts)" directionality="2" translationMW="37384.08" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="3496-4446" color="#993366" 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;rep101&lt;/i&gt; encoding an A56V mutation&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/6327996&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/6327996&quot;&gt;Armstrong et al., 1984&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKERTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFSSEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANIEISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQVELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKFLSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI*"/></Q></Feature><Feature recentID="9" name="Shine-Dalgarno sequence" type="RBS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1227-1235" 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="10" name="Gam" directionality="1" translationMW="16346.73" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1244-1660" color="#ff0000" 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;gam&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;protects the double-stranded DNA substrate for recombination&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;inhibitor of the host RecBCD nuclease in the&amp;nbsp;Î» Red system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYYIQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITRAFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV*"/></Q></Feature><Feature recentID="11" name="Beta" directionality="1" translationMW="29689.57" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1666-2451" color="#ff0000" 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;bet&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;mediates pairing of a single-stranded DNA with a complementary target&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;single-stranded DNA binding recombinase in the&amp;nbsp;Î» Red system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFIALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESCTCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLAFGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLPLCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA*"/></Q></Feature><Feature recentID="12" name="Exo" directionality="1" translationMW="25909.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2448-3128" color="#ff0000" 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;exo&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;creates 3' single-stranded DNA tails&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;5' to 3' double-stranded DNA exonuclease in the&amp;nbsp;Î» Red system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPRSGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESPIIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMWVTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFGEQWR*"/></Q></Feature><Feature recentID="13" name="Î» tL3 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="3129-3373" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator &lt;i&gt;t&lt;/i&gt;&lt;sub&gt;L3&lt;/sub&gt; from phage&amp;nbsp;Î»&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T1 terminator" type="terminator" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5740-5786" 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="15" name="Myc" directionality="1" translationMW="1203.34" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5517" maxFusedRunOn="5517" detectionMode="exactProteinMatch"><Segment range="5452-5481" 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;Myc (human c-Myc proto-oncogene) epitope tag&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="EQKLISEEDL"/></Q></Feature><Feature recentID="16" name="6xHis" directionality="1" translationMW="840.86" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5517" maxFusedRunOn="5517" detectionMode="exactProteinMatch"><Segment range="5497-5514" 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;6xHis affinity tag&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="HHHHHH"/></Q></Feature></Features>
  å<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="pBAD Forward" sequence="ATGCCATAGCATTTTTATCC" description="For vectors with E. coli araBAD promoter, forward primer"><BindingSite location="1122-1141" boundStrand="0" annealedBases="ATGCCATAGCATTTTTATCC" meltingTemperature="51"><Component hybridizedRange="1122-1141" bases="ATGCCATAGCATTTTTATCC"/></BindingSite><BindingSite simplified="1" location="1122-1141" boundStrand="0" annealedBases="ATGCCATAGCATTTTTATCC" meltingTemperature="51"><Component hybridizedRange="1122-1141" bases="ATGCCATAGCATTTTTATCC"/></BindingSite></Primer><Primer recentID="1" name="pBAD Reverse" sequence="GATTTAATCTGTATCAGG" description="For vectors with E. coli araBAD promoter, reverse primer"><BindingSite location="5569-5586" boundStrand="1" annealedBases="GATTTAATCTGTATCAGG" meltingTemperature="45"><Component hybridizedRange="5569-5586" bases="GATTTAATCTGTATCAGG"/></BindingSite><BindingSite simplified="1" location="5569-5586" boundStrand="1" annealedBases="GATTTAATCTGTATCAGG" meltingTemperature="45"><Component hybridizedRange="5569-5586" bases="GATTTAATCTGTATCAGG"/></BindingSite></Primer></Primers>
  ›<Notes>
<UUID>9edbd218-31f7-4383-a454-5b999b69b853</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pKDsgRNA-p15</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>Arabinose inducible lambda red and anhydrotetracycline inducible sgRNA expression that targets pCas9 plasmid.</Description>
<Created UTC="20:50:38">2021.5.12</Created>
<LastModified UTC="20:50:38">2021.5.12</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
<References>
<Reference volume="5:15096. doi" type="Journal Article" journalName="Sci Rep." title="The no-SCAR (Scarless Cas9 Assisted Recombineering) system for genome editing in Escherichia coli." journal="Sci Rep. 2015 Oct 14;5:15096. doi: 10.1038/srep15096." authors="Reisch CR, Prather KL" pubMedID="26463009" date="2015-10-14" pages=" 10.1038/srep15096"/>
</References>
</Notes>
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   )<?xml version="1.0"?><CustomEnzymeSets/>
