LOCUS pTR_scU6-Ai9 5821 bp ds-DNA circular 20-MAY-2021 DEFINITION . FEATURES Location/Qualifiers misc_feature 1..115 /label="ITR1" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd promoter 121..361 /label="U6 promoter" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 misc_feature 370..390 /label="Ai9-Sa-gRNA1_With G" /ApEinfo_revcolor=#3366ff /ApEinfo_fwdcolor=#3366ff misc_feature 391..404 /label="crRNA Direct Repeat Portion +1 to +14" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac misc_feature 391..471 /label="sgRNA scaffold" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature 405..408 /label="synthetic tetraloop" /ApEinfo_revcolor=#000000 /ApEinfo_fwdcolor=#000000 misc_feature 409..471 /label="TracrRNA" /ApEinfo_revcolor=#ff0000 /ApEinfo_fwdcolor=#ff0000 misc_feature 472..472 /label="Additional T introduced in gblock" /ApEinfo_revcolor=#ff0000 /ApEinfo_fwdcolor=#ff0000 primer_bind complement(505..524) /label="T3" /ApEinfo_revcolor=#008b8b /ApEinfo_fwdcolor=#008b8b promoter 531..771 /label="U6 promoter" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 misc_feature 780..800 /label="Ai9-Sa-gRNA2_With G" /ApEinfo_revcolor=#3366ff /ApEinfo_fwdcolor=#3366ff misc_feature 801..814 /label="crRNA Direct Repeat Portion +1 to +14" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac misc_feature 801..881 /label="sgRNA scaffold" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature 815..818 /label="synthetic tetraloop" /ApEinfo_revcolor=#000000 /ApEinfo_fwdcolor=#000000 misc_feature 819..881 /label="TracrRNA" /ApEinfo_revcolor=#ff0000 /ApEinfo_fwdcolor=#ff0000 misc_feature 882..882 /label="Additional T introduced in gblock" /ApEinfo_revcolor=#ff0000 /ApEinfo_fwdcolor=#ff0000 misc_feature 1104..1823 /label="EGFP" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 misc_feature 1841..2098 /label="BGH+PolyA" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e LTR 2105..2234 /label="ITR2" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd CDS 2236..2304 /label="LacZ alpha" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc rep_origin 2605..3060 /label="M13 origin" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 rep_origin 2622..2928 /label="F1 ori" /ApEinfo_revcolor=#0080c0 /ApEinfo_fwdcolor=#0080c0 CDS 4167..4826 /label="AmpR" /ApEinfo_revcolor=#ff4242 /ApEinfo_fwdcolor=#ff4242 rep_origin 4924..5606 /label="ColE1 origin" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 ORIGIN 1 CAGCTGCGCG CTCGCTCGCT CACTGAGGCC GCCCGGGCAA AGCCCGGGCG TCGGGCGACC 61 TTTGGTCGCC CGGCCTCAGT GAGCGAGCGA GCGCGCAGAG AGGGAGTGGG GTTCGGTACC 121 GAGGGCCTAT TTCCCATGAT TCCTTCATAT TTGCATATAC GATACAAGGC TGTTAGAGAG 181 ATAATTAGAA TTAATTTGAC TGTAAACACA AAGATATTAG TACAAAATAC GTGACGTAGA 241 AAGTAATAAT TTCTTGGGTA GTTTGCAGTT TTAAAATTAT GTTTTAAAAT GGACTATCAT 301 ATGCTTACCG TAACTTGAAA GTATTTCGAT TTCTTGGCTT TATATATCTT GTGGAAAGGA 361 CGAAACACCG CTCTAGAGTC GCAGATCCTC gttttagtac tctggaaaca gaatctacta 421 aaacaaggca aaatgccgtg tttatctcgt caacttgttg gcgagatttt ttGCGGCCGC 481 CCGCGGTGGA GCTCCAGCTT TTGTTCCCTT TAGTGAGGGT TAATCCTAGG GAGGGCCTAT 541 TTCCCATGAT TCCTTCATAT TTGCATATAC GATACAAGGC TGTTAGAGAG ATAATTAGAA 601 TTAATTTGAC TGTAAACACA AAGATATTAG TACAAAATAC GTGACGTAGA AAGTAATAAT 661 TTCTTGGGTA GTTTGCAGTT TTAAAATTAT GTTTTAAAAT GGACTATCAT ATGCTTACCG 721 TAACTTGAAA GTATTTCGAT TTCTTGGCTT TATATATCTT GTGGAAAGGA CGAAACACCG 781 ACGAAGTTAT ATTAAGGGTT gttttagtac tctggaaaca gaatctacta aaacaaggca 841 aaatgccgtg tttatctcgt caacttgttg gcgagatttt ttTCTAGagg atccggtact 901 cgaggaactg aaaaaccaga aagttaactg gtaagtttag tctttttgtc ttttatttca 961 ggtcccggat ccggtggtgg tgcaaatcaa agaactgctc ctcagtggat gttgccttta 1021 cttctaggcc tgtacggaag tgttacttct gctctaaaag ctgcggaatt gtacccgcgg 1081 cccgggatcc aCCGGTCGCC ACCATGGTGA GCAAGGGCGA GGAGCTGTTC ACCGGGGTGG 1141 TGCCCATCCT GGTCGAGCTG GACGGCGACG TAAACGGCCA CAAGTTCAGC GTGTCCGGCG 1201 AGGGCGAGGG CGATGCCACC TACGGCAAGC TGACCCTGAA GTTCATCTGC ACCACCGGCA 1261 AGCTGCCCGT GCCCTGGCCC ACCCTCGTGA CCACCCTGAC CTACGGCGTG CAGTGCTTCA 1321 GCCGCTACCC CGACCACATG AAGCAGCACG ACTTCTTCAA GTCCGCCATG CCCGAAGGCT 1381 ACGTCCAGGA GCGCACCATC TTCTTCAAGG ACGACGGCAA CTACAAGACC CGCGCCGAGG 1441 TGAAGTTCGA GGGCGACACC CTGGTGAACC GCATCGAGCT GAAGGGCATC GACTTCAAGG 1501 AGGACGGCAA CATCCTGGGG CACAAGCTGG AGTACAACTA CAACAGCCAC AACGTCTATA 1561 TCATGGCCGA CAAGCAGAAG AACGGCATCA AGGTGAACTT CAAGATCCGC CACAACATCG 1621 AGGACGGCAG CGTGCAGCTC GCCGACCACT ACCAGCAGAA CACCCCCATC GGCGACGGCC 1681 CCGTGCTGCT GCCCGACAAC CACTACCTGA GCACCCAGTC CGCCCTGAGC AAAGACCCCA 1741 ACGAGAAGCG CGATCACATG GTCCTGCTGG AGTTCGTGAC CGCCGCCGGG ATCACTCTCG 1801 GCATGGACGA GCTGTACAAG TAAAGCGGCC GCGCGGATCC CTCGACTGTG CCTTCTAGTT 1861 GCCAGCCATC TGTTGTTTGC CCCTCCCCCG TGCCTTCCTT GACCCTGGAA GGTGCCACTC 1921 CCACTGTCCT TTCCTAATAA AATGAGGAAA TTGCATCGCA TTGTCTGAGT AGGTGTCATT 1981 CTATTCTGGG GGGTGGGGTG GGGCAGGACA GCAAGGGGGA GGATTGGGAA GACAACAGCA 2041 GGCATGCTGG GGATGCGGTG GGCTCTATGG CTTCTGAGGC GGAAAGAACC AGCTTTGGAC 2101 GCGTAGGAAC CCCTAGTGAT GGAGTTGGCC ACTCCCTCTC TGCGCGCTCG CTCGCTCACT 2161 GAGGCCGGGC GACCAAAGGT CGCCCGACGC CCGGGCTTTG CCCGGGCGGC CTCAGTGAGC 2221 GAGCGAGCGC GCAGCTGGCG TAATAGCGAA GAGGCCCGCA CCGATCGCCC TTCCCAACAG 2281 TTGCGCAGCC TGAATGGCGA ATGGCGATTC CGTTGCAATG GCTGGCGGTA ATATTGTTCT 2341 GGATATTACC AGCAAGGCCG ATAGTTTGAG TTCTTCTACT CAGGCAAGTG ATGTTATTAC 2401 TAATCAAAGA AGTATTGCGA CAACGGTTAA TTTGCGTGAT GGACAGACTC TTTTACTCGG 2461 TGGCCTCACT GATTATAAAA ACACTTCTCA GGATTCTGGC GTACCGTTCC TGTCTAAAAT 2521 CCCTTTAATC GGCCTCCTGT TTAGCTCCCG CTCTGATTCT AACGAGGAAA GCACGTTATA 2581 CGTGCTCGTC AAAGCAACCA TAGTACGCGC CCTGTAGCGG CGCATTAAGC GCGGCGGGTG 2641 TGGTGGTTAC GCGCAGCGTG ACCGCTACAC TTGCCAGCGC CCTAGCGCCC GCTCCTTTCG 2701 CTTTCTTCCC TTCCTTTCTC GCCACGTTCG CCGGCTTTCC CCGTCAAGCT CTAAATCGGG 2761 GGCTCCCTTT AGGGTTCCGA TTTAGTGCTT TACGGCACCT CGACCCCAAA AAACTTGATT 2821 AGGGTGATGG TTCACGTAGT GGGCCATCGC CCTGATAGAC GGTTTTTCGC CCTTTGACGT 2881 TGGAGTCCAC GTTCTTTAAT AGTGGACTCT TGTTCCAAAC TGGAACAACA CTCAACCCTA 2941 TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT TGGTTAAAAA 3001 ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG TTTACAATTT 3061 AAATATTTGC TTATACAATC TTCCTGTTTT TGGGGCTTTT CTGATTATCA ACCGGGGTAC 3121 ATATGATTGA CATGCTAGTT TTACGATTAC CGTTCATCGA TTCTCTTGTT TGCTCCAGAC 3181 TCTCAGGCAA TGACCTGATA GCCTTTGTAG AGACCTCTCA AAAATAGCTA CCCTCTCCGG 3241 CATGAATTTA TCAGCTAGAA CGGTTGAATA TCATATTGAT GGTGATTTGA CTGTCTCCGG 3301 CCTTTCTCAC CCGTTTGAAT CTTTACCTAC ACATTACTCA GGCATTGCAT TTAAAATATA 3361 TGAGGGTTCT AAAAATTTTT ATCCTTGCGT TGAAATAAAG GCTTCTCCCG CAAAAGTATT 3421 ACAGGGTCAT AATGTTTTTG GTACAACCGA TTTAGCTTTA TGCTCTGAGG CTTTATTGCT 3481 TAATTTTGCT AATTCTTTGC CTTGCCTGTA TGATTTATTG GATGTTGGAA TTCCTGATGC 3541 GGTATTTTCT CCTTACGCAT CTGTGCGGTA TTTCACACCG CATATGGTGC ACTCTCAGTA 3601 CAATCTGCTC TGATGCCGCA TAGTTAAGCC AGCCCCGACA CCCGCCAACA CCCGCTGACG 3661 CGCCCTGACG GGCTTGTCTG CTCCCGGCAT CCGCTTACAG ACAAGCTGTG ACCGTCTCCG 3721 GGAGCTGCAT GTGTCAGAGG TTTTCACCGT CATCACCGAA ACGCGCGAGA CGAAAGGGCC 3781 TCGTGATACG CCTATTTTTA TAGGTTAATG TCATGATAAT AATGGTTTCT TAGACGTCAG 3841 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT 3901 CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA 3961 GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT 4021 GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT 4081 TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT 4141 TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG 4201 TATTATCCCG TATTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA 4261 ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA 4321 GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA 4381 CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA 4441 CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA 4501 CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA 4561 CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC 4621 TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC 4681 GTGGGTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG 4741 TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA 4801 TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT 4861 AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA 4921 ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG 4981 AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA 5041 CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT 5101 TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTCCTT CTAGTGTAGC 5161 CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA 5221 TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA 5281 GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC 5341 CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CTATGAGAAA 5401 GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA 5461 CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG 5521 GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC 5581 TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG 5641 CTCACATGTT CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG 5701 AGTGAGCTGA TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG 5761 AAGCGGAAGA GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT 5821 G //