LOCUS Exported 8307 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR system used to genome editing in Mycobacterium smegmatis, expresses Fn CPf1. ACCESSION . VERSION . KEYWORDS pSBY1_FnCpf1cg SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8307) AUTHORS Sun B, Yang J, Yang S, Ye RD, Chen D, Jiang Y TITLE A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing System of Mycobacterium smegmatis. JOURNAL Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588. Epub 2018 Aug 6. PUBMED 30039929 REFERENCE 2 (bases 1 to 8307) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8307 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(128..147) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 139..954 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(212..231) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1285..1873 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1774..1793 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 8239..8285 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 gctagccacc tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa tctctgatgt 61 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac 121 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc gaggccgcga 181 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg 241 caatcaggtg cgacaatcta tcgcttgtat gggaagcccc atgcgccaga gttgtttctg 301 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg 361 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca 421 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct 481 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt 541 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca 601 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct 661 gttgaacaag tctggaaaga aatgcataat cttttgccat tctcaccgga ttcagtcgtc 721 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt 781 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac 841 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat 901 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa 961 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt 1021 gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg 1081 cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag 1141 ctctcaccaa ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt 1201 atgagtcagc aacaccttct tcacgaggca gacctcacta gttccatgag cgtcagaccc 1261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 1321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 1381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 1441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 1501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 1561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 1621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 1681 agaaagcgcc acgcttcccg aggggagaaa ggcggacagg tatccggtaa gcggcagggt 1741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 1801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 1861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 1921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 1981 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcaacg cgtgagccca 2041 ccagctccgt aagttcgggc gctgtgtggc tcgtacccgc gcattcaggc ggcagggggt 2101 ctaacgggtc taaggcggcg tgtacggccg ccacagcggc tctcagcggc ccggaaacgt 2161 cctcgaaacg acgcatgtgt tcctcctggt tggtacaggt ggttgggggt gctcggctgt 2221 cgctggtgtt ccaccaccag ggctcgacgg gagagcgggg gagtgtgcag ttgtggggtg 2281 gcccctcagc gaaatatctg acttggagct cgtgtcggac catacaccgg tgattaatcg 2341 tggtctacta ccaagcgtga gccacgtcgc cgacgaattt gagcagctct ggctgccgta 2401 ctggccgctg gcaagcgacg atctgctcga ggggatctac cgccaaagcc gcgcgtcggc 2461 cctaggccgc cggtacatcg aggcgaaccc aacagcgctg gcaaacctgc tggtcgtgga 2521 cgtagaccat ccagacgcag cgctccgagc gctcagcgcc cgggggtccc atccgctgcc 2581 caacgcgatc gtgggcaatc gcgccaacgg ccacgcacac gcagtgtggg cactcaacgc 2641 ccctgttcca cgcaccgaat acgcgcggcg taagccgctc gcatacatgg cggcgtgcgc 2701 cgaaggcctt cggcgcgccg tcgatggcga ccgcagttac tcaggcctca tgaccaaaaa 2761 ccccggccac atcgcctggg aaacggaatg gctccactca gatctctaca cactcagcca 2821 catcgaggcc gagctcggcg cgaacatgcc accgccgcgc tggcgtcagc agaccacgta 2881 caaagcggct ccgacgccgc tagggcggaa ttgcgcactg ttcgattccg tcaggttgtg 2941 ggcctatcgt cccgccctca tgcggatcta cctgccgacc cggaacgtgg acggactcgg 3001 ccgcgcgatc tatgccgagt gccacgcgcg aaacgccgaa tttccgtgca acgacgtgtg 3061 tcccggaccg ctaccggaca gcgaggtccg cgccatcgcc aacagcattt ggcgttggat 3121 cacaaccaag tcgcgcattt gggcggacgg gatcgtggtc tacgaggcca cactcagtgc 3181 gcgccagtcg gccatctcgc ggaagggcgc agcagcgcgc acggcggcga gcacagttgc 3241 gcggcgcgca aagtccgcgt cagccatgga ggcattgcta tgagcgacgg ctacagcgac 3301 ggctacaacc ggcagccgac tgtccgcaaa aagcggcgcg tgaccgccgc cgaaggcgct 3361 cgaatcaccg gactatccga acgccacgtc gtccggctcg tggcgcagga acgcagcgag 3421 tggctcgccg agcaggctgc acgccgcgaa cgcatccgcg cctatcacga cgacgagggc 3481 cactcttggc cgcaaacggc caaacatttc gggctgcatc tggacaccgt taagcgactc 3541 ggctatcggg cgaggaaaga gcgtgcggca gaacaggaag cggctcaaaa ggcccacaac 3601 gaagccgaca atccaccgct gttctaacgc aattggggag cgggtgtcgc gggggttccg 3661 tggggggttc cgttgcaacg ggtcggacag gtaaaagtcc tggtagacgc tagttttctg 3721 gtttgggcca tgcctgtctc gttgcgtgtt tcgttgcgcc cgttttgaat accagccaga 3781 cgagacgggg ttctacgaat cttggtcgat accaagccat ttccgctgaa tatcggggag 3841 ctcaccgcca gaatcggtgg ttgtggtgat gtacgtggcg aactccgttg tagtgcctgt 3901 ggtggcatcc gtggcgcggc cgcggtacca gatctttaaa tctagacggt gaccacaacg 3961 cgcccgcttt gatcggggac gtctgcggcc gaccatttac gggtcttgtt gtcgttggcg 4021 gtcatgggcc gaacatactc acccggatcg gagggccgag gacaaggtcg aacgaggggc 4081 atgacccggt gcggggcttc ttgcactcgg cataggcgag tgctaagaat aacgttggca 4141 ctcgcgaccg gtgagtgcta ggtcgggacg gtgaggccag gcccgtcgtc gcagcgagtg 4201 gcagcgagga caacttgagc cgtccgtcgc gggcactgcg cccggccagc gtaagtagcg 4261 gggttgccgt cacccggtga cccccgtttc atccccgatc cggaggaatc acttcgcaat 4321 gtccatctac caagagtttg tgaataaata ctccctgtcc aagaccctcc gttttgagct 4381 gatcccccaa ggcaagaccc tcgaaaacat caaggcacgc ggcctcatcc tggatgacga 4441 aaagcgcgct aaggattaca agaaggcaaa gcagatcatc gacaagtacc accagttctt 4501 catcgaagag atcctgtcct ccgtgtgcat ctccgaggac ctgctccaga actactccga 4561 tgtctacttc aagctcaaga agtccgatga cgataacctg cagaaggact tcaagtccgc 4621 taaggatacc atcaagaagc agatctccga atacatcaag gattccgaga agttcaagaa 4681 cctcttcaac cagaacctga tcgacgcaaa gaagggccag gaatccgatc tcatcctgtg 4741 gctcaagcag tccaaggata acggcatcga gctcttcaag gccaactccg acatcaccga 4801 catcgatgaa gctctggaga tcatcaagtc cttcaagggc tggaccacct acttcaaggg 4861 cttccacgaa aaccgcaaga acgtgtactc ctccaacgat atcccaacct ctatcatcta 4921 ccgcatcgtc gacgataacc tgccaaagtt cctcgaaaac aaggcaaagt acgagtccct 4981 gaaggataag gccccagaag ctatcaacta cgagcagatc aagaaggacc tggccgaaga 5041 gctcaccttc gacatcgatt acaagacctc tgaagtgaac cagcgcgtct tctccctcga 5101 tgaagtgttc gagatcgcca acttcaacaa ctacctgaac cagtccggca tcaccaagtt 5161 caacaccatc atcggcggca agttcgtcaa cggcgaaaac accaagcgca agggcatcaa 5221 cgagtacatc aacctctact cccagcagat caacgataag accctgaaga agtacaagat 5281 gtccgtgctc ttcaagcaga tcctgtccga caccgaatcc aagtccttcg tcatcgacaa 5341 gctggaggac gattccgatg tggtcaccac catgcagtcc ttctacgaac agatcgcagc 5401 cttcaagacc gtggaagaga agtccatcaa ggagaccctc tccctgctct tcgacgatct 5461 gaaggctcag aagctggatc tctccaagat ctacttcaag aacgacaagt ccctgaccga 5521 tctctcccag caggtcttcg acgattactc cgtgatcggc accgcagtcc tggaatacat 5581 cacccagcag atcgccccaa agaacctcga taacccatcc aagaaggaac aggagctgat 5641 cgccaagaag accgaaaagg ctaagtacct gtccctcgag accatcaagc tggctctcga 5701 agagttcaac aagcaccgcg acatcgataa gcagtgccgc ttcgaagaga tcctcgcaaa 5761 cttcgctgca atcccaatga tcttcgacga aatcgcacag aacaaggata acctggccca 5821 gatctccatc aagtaccaga accagggcaa gaaggatctg ctccaggcct ccgctgagga 5881 cgatgtgaag gcaatcaagg acctgctcga tcagaccaac aacctgctcc acaagctgaa 5941 gatcttccac atctcccagt ccgaagacaa ggccaacatc ctcgacaagg atgagcactt 6001 ctacctggtg ttcgaagagt gctacttcga actcgctaac atcgtcccac tgtacaacaa 6061 gatccgcaac tacatcaccc agaagccata ctccgatgaa aagttcaagc tcaacttcga 6121 gaactccacc ctggcaaacg gctgggacaa gaacaaggaa ccagataaca ccgccatcct 6181 cttcatcaag gacgataagt actacctggg cgtgatgaac aagaagaaca acaagatctt 6241 cgacgataag gccatcaagg aaaacaaggg cgagggctac aagaagatcg tgtacaagct 6301 gctcccaggc gctaacaaga tgctcccaaa ggtcttcttc tccgcaaagt ccatcaagtt 6361 ctacaaccca tccgaagata tcctgcgcat ccgcaaccac tccacccaca ccaagaacgg 6421 ctccccacag aagggctacg aaaagttcga gttcaacatc gaagactgcc gcaagttcat 6481 cgatttctac aagcagtcca tctccaagca cccagagtgg aaggacttcg gcttccgctt 6541 ctccgatacc cagcgctaca actccatcga tgaattctac cgcgaagtgg agaaccaggg 6601 ctacaagctg accttcgaaa acatctccga gtcctacatc gattccgtgg tcaaccaggg 6661 caagctgtac ctcttccaga tctacaacaa ggacttctcc gcttactcca agggccgccc 6721 aaacctgcac accctctact ggaaggcact cttcgacgaa cgcaacctgc aggatgtggt 6781 ctacaagctc aacggcgaag cagagctgtt ctaccgcaag cagtccatcc caaagaagat 6841 cacccaccca gccaaggaag caatcgccaa caagaacaag gataacccaa agaaggaatc 6901 cgtgttcgag tacgacctga tcaaggataa gcgcttcacc gaggacaagt tcttcttcca 6961 ctgcccaatc accatcaact tcaagtcctc cggcgccaac aagttcaacg atgaaatcaa 7021 cctgctcctg aaggagaagg ctaacgacgt gcacatcctg tccatcgatc gcggcgaacg 7081 ccacctcgcc tactacaccc tggtcgacgg caagggcaac atcatcaagc aggacacctt 7141 caacatcatc ggcaacgatc gcatgaagac caactaccac gacaagctgg ccgctatcga 7201 gaaggaccgc gattccgctc gcaaggattg gaagaagatc aacaacatca aggaaatgaa 7261 ggaaggctac ctctcccagg tggtccacga aatcgctaag ctggtgatcg agtacaacgc 7321 aatcgtggtc ttcgaagacc tgaacttcgg cttcaagcgc ggccgcttca aggtggagaa 7381 gcaggtctac cagaagctgg aaaagatgct catcgagaag ctgaactacc tcgtgttcaa 7441 ggacaacgaa ttcgataaga ccggcggcgt cctccgtgca taccagctga ccgccccatt 7501 cgagaccttc aagaagatgg gcaagcagac cggcatcatc tactacgtgc cagctggctt 7561 cacctctaag atctgcccag tgaccggctt cgtcaaccag ctctacccaa agtacgaatc 7621 cgtctccaag tcccaggagt tcttctccaa gttcgacaag atctgctaca acctggataa 7681 gggctacttc gaattctcct tcgactacaa gaacttcggc gataaggcag ccaagggcaa 7741 gtggaccatc gcatccttcg gctcccgcct catcaacttc cgcaactccg acaagaacca 7801 caactgggat acccgcgaag tgtacccaac caaggaactg gagaagctcc tgaaggatta 7861 ctccatcgaa tacggccacg gcgagtgcat caaggctgca atctgcggcg aatccgacaa 7921 gaagttcttc gcaaagctga cctctgtgct caacaccatc ctgcagatgc gcaactccaa 7981 gaccggcacc gagctggatt acctcatctc cccagtggcc gacgtcaacg gcaacttctt 8041 cgattcccgc caggctccaa agaacatgcc acaggacgct gatgcaaacg gcgcctacca 8101 catcggtctg aagggtctca tgctcctggg tcgcatcaag aacaaccagg aaggcaagaa 8161 gctgaatctc gtcattaaga acgaagaata ctttgaattt gtccagaacc gcaataacta 8221 agatcgactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt 8281 tttatgccat catggccgcg gtgatca //