LOCUS       sequence-201573-       10039 bp    DNA     circular SYN 14-APR-2025
DEFINITION  CRISPR system used to genome editing in Mycobacterium smegmatis, 
            expresses Fn Cpf1 and crRNA targeting ligD and hygromysin resistance
            gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10039)
  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 10039)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            J."; date: "2018-09"; volume: "13(9):e1700588. doi"; pages: " 
            10.1002/biot.201700588. Epub 2018 Aug 6"
COMMENT     Sequence Label: pSBY3_ligD-HygR
FEATURES             Location/Qualifiers
     source          1..10039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..53)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(145..230)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(438..457)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(522..541)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3017..3605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3506..3525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      9971..10017
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat
       61 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc
      121 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
      181 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt aaatcaccgt
      241 tgcgtgtgac gtgctatcta caacagtaga aattcggatc cattatacct aggactgagc
      301 tagctgtcaa tctagccacc tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa
      361 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
      421 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
      481 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
      541 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccc atgcgccaga
      601 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
      661 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
      721 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
      781 agaatatcct gattcaggtg aaaatagatg atcgaggatg ccagggcctt tcacgcccgc
      841 cgctgctgag cgtccgccgc cgggcggccc ggggcgtcag gcgccggggg cggtgtccgg
      901 cggcccccag aggaactgcg ccagttcctc cggatcggtg aagccggaga gatccagcgg
      961 ggtctcctcg aacacctcga agtcgtgcag gaaggtgaag gcgagcagtt cgctggcgaa
     1021 gtcctcggtc cgcttccact gcgccccgtc gagcagcgcg gccaggatct cgcggtcgcc
     1081 ccggaaggcg ttgagatgca gttgcaccag gctgtagcgg gagtctcccg catagacgtc
     1141 ggtgaagtcg acgatcccgg tgacctcggt cgcggccagg tccacgaaga tgttggtccc
     1201 gtgcaggtcg ccgtggacga accggggttc gcggccggcc agcagcgtgt ccacgtccgg
     1261 cagccagtcc tccaggcggt ccagcagccg gggcgagagg tagccccacc cgcggtggtc
     1321 ctcgacggtc gccgcgcggc gttcccgcag cagttccggg aagacctcgg aatggggggt
     1381 gagcacggtg ttcccggtca gcggcaccct gtgcagccgg ccgagcaccc ggccgagttc
     1441 gcgggccagg gcgagcagcg cgttccggtc ggtcgtgccg tccatcgcgg accgccaggt
     1501 ggtgccggtc atccggctca tcaccaggta gggccacggc caggctccgg tgccgggccg
     1561 cagctcgccg cggccgagga ggcggggcac cggcaccggg gcgtccgcca ggaccgcgta
     1621 cgcctccgac tccgacgcga ggctctccgg accgcaccag tgctcgccga acagcttgat
     1681 caccgggccg ggctcgccga ccagtacggg gttggtgctc tcgccgggca cccgcagcac
     1741 cggcggcacc ggcagcccga gctcctccag ggctcggcgg gccagcggct cccagaattc
     1801 ctggtcgttc cgcaggctcg cgtaggaatc atccgaatca atacggtcga gaagtaacag
     1861 ggattcttgt gtcacagcgg acctctattc acagggtacg ggccggctta attccgcacg
     1921 gccggtcgcg acacggcctg tccgcaccgc ggatcaggcg ttgacgatga cgggctggtc
     1981 ggccacgtcg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2041 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2221 gatcaaagat tgttgatcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2281 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2341 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2401 agtctggaaa gaaatgcata atcttttgcc attctcaccg gattcagtcg tcactcatgg
     2461 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2521 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2581 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2641 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2701 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata
     2761 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     2821 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcacc
     2881 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca
     2941 gcaacacctt cttcacgagg cagacctcac tagttccatg agcgtcagac cccgtagaaa
     3001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3421 ccacgcttcc cgaggggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3721 gagctgatac cgctcgccgc agccgaacga ccgagcgcaa cgcgtgagcc caccagctcc
     3781 gtaagttcgg gcgctgtgtg gctcgtaccc gcgcattcag gcggcagggg gtctaacggg
     3841 tctaaggcgg cgtgtacggc cgccacagcg gctctcagcg gcccggaaac gtcctcgaaa
     3901 cgacgcatgt gttcctcctg gttggtacag gtggttgggg gtgctcggct gtcgctggtg
     3961 ttccaccacc agggctcgac gggagagcgg gggagtgtgc agttgtgggg tggcccctca
     4021 gcgaaatatc tgacttggag ctcgtgtcgg accatacacc ggtgattaat cgtggtctac
     4081 taccaagcgt gagccacgtc gccgacgaat ttgagcagct ctggctgccg tactggccgc
     4141 tggcaagcga cgatctgctc gaggggatct accgccaaag ccgcgcgtcg gccctaggcc
     4201 gccggtacat cgaggcgaac ccaacagcgc tggcaaacct gctggtcgtg gacgtagacc
     4261 atccagacgc agcgctccga gcgctcagcg cccgggggtc ccatccgctg cccaacgcga
     4321 tcgtgggcaa tcgcgccaac ggccacgcac acgcagtgtg ggcactcaac gcccctgttc
     4381 cacgcaccga atacgcgcgg cgtaagccgc tcgcatacat ggcggcgtgc gccgaaggcc
     4441 ttcggcgcgc cgtcgatggc gaccgcagtt actcaggcct catgaccaaa aaccccggcc
     4501 acatcgcctg ggaaacggaa tggctccact cagatctcta cacactcagc cacatcgagg
     4561 ccgagctcgg cgcgaacatg ccaccgccgc gctggcgtca gcagaccacg tacaaagcgg
     4621 ctccgacgcc gctagggcgg aattgcgcac tgttcgattc cgtcaggttg tgggcctatc
     4681 gtcccgccct catgcggatc tacctgccga cccggaacgt ggacggactc ggccgcgcga
     4741 tctatgccga gtgccacgcg cgaaacgccg aatttccgtg caacgacgtg tgtcccggac
     4801 cgctaccgga cagcgaggtc cgcgccatcg ccaacagcat ttggcgttgg atcacaacca
     4861 agtcgcgcat ttgggcggac gggatcgtgg tctacgaggc cacactcagt gcgcgccagt
     4921 cggccatctc gcggaagggc gcagcagcgc gcacggcggc gagcacagtt gcgcggcgcg
     4981 caaagtccgc gtcagccatg gaggcattgc tatgagcgac ggctacagcg acggctacaa
     5041 ccggcagccg actgtccgca aaaagcggcg cgtgaccgcc gccgaaggcg ctcgaatcac
     5101 cggactatcc gaacgccacg tcgtccggct cgtggcgcag gaacgcagcg agtggctcgc
     5161 cgagcaggct gcacgccgcg aacgcatccg cgcctatcac gacgacgagg gccactcttg
     5221 gccgcaaacg gccaaacatt tcgggctgca tctggacacc gttaagcgac tcggctatcg
     5281 ggcgaggaaa gagcgtgcgg cagaacagga agcggctcaa aaggcccaca acgaagccga
     5341 caatccaccg ctgttctaac gcaattgggg agcgggtgtc gcgggggttc cgtggggggt
     5401 tccgttgcaa cgggtcggac aggtaaaagt cctggtagac gctagttttc tggtttgggc
     5461 catgcctgtc tcgttgcgtg tttcgttgcg cccgttttga ataccagcca gacgagacgg
     5521 ggttctacga atcttggtcg ataccaagcc atttccgctg aatatcgggg agctcaccgc
     5581 cagaatcggt ggttgtggtg atgtacgtgg cgaactccgt tgtagtgcct gtggtggcat
     5641 ccgtggcgcg gccgcggtac cagatcttta aatctagacg gtgaccacaa cgcgcccgct
     5701 ttgatcgggg acgtctgcgg ccgaccattt acgggtcttg ttgtcgttgg cggtcatggg
     5761 ccgaacatac tcacccggat cggagggccg aggacaaggt cgaacgaggg gcatgacccg
     5821 gtgcggggct tcttgcactc ggcataggcg agtgctaaga ataacgttgg cactcgcgac
     5881 cggtgagtgc taggtcggga cggtgaggcc aggcccgtcg tcgcagcgag tggcagcgag
     5941 gacaacttga gccgtccgtc gcgggcactg cgcccggcca gcgtaagtag cggggttgcc
     6001 gtcacccggt gacccccgtt tcatccccga tccggaggaa tcacttcgca atgtccatct
     6061 accaagagtt tgtgaataaa tactccctgt ccaagaccct ccgttttgag ctgatccccc
     6121 aaggcaagac cctcgaaaac atcaaggcac gcggcctcat cctggatgac gaaaagcgcg
     6181 ctaaggatta caagaaggca aagcagatca tcgacaagta ccaccagttc ttcatcgaag
     6241 agatcctgtc ctccgtgtgc atctccgagg acctgctcca gaactactcc gatgtctact
     6301 tcaagctcaa gaagtccgat gacgataacc tgcagaagga cttcaagtcc gctaaggata
     6361 ccatcaagaa gcagatctcc gaatacatca aggattccga gaagttcaag aacctcttca
     6421 accagaacct gatcgacgca aagaagggcc aggaatccga tctcatcctg tggctcaagc
     6481 agtccaagga taacggcatc gagctcttca aggccaactc cgacatcacc gacatcgatg
     6541 aagctctgga gatcatcaag tccttcaagg gctggaccac ctacttcaag ggcttccacg
     6601 aaaaccgcaa gaacgtgtac tcctccaacg atatcccaac ctctatcatc taccgcatcg
     6661 tcgacgataa cctgccaaag ttcctcgaaa acaaggcaaa gtacgagtcc ctgaaggata
     6721 aggccccaga agctatcaac tacgagcaga tcaagaagga cctggccgaa gagctcacct
     6781 tcgacatcga ttacaagacc tctgaagtga accagcgcgt cttctccctc gatgaagtgt
     6841 tcgagatcgc caacttcaac aactacctga accagtccgg catcaccaag ttcaacacca
     6901 tcatcggcgg caagttcgtc aacggcgaaa acaccaagcg caagggcatc aacgagtaca
     6961 tcaacctcta ctcccagcag atcaacgata agaccctgaa gaagtacaag atgtccgtgc
     7021 tcttcaagca gatcctgtcc gacaccgaat ccaagtcctt cgtcatcgac aagctggagg
     7081 acgattccga tgtggtcacc accatgcagt ccttctacga acagatcgca gccttcaaga
     7141 ccgtggaaga gaagtccatc aaggagaccc tctccctgct cttcgacgat ctgaaggctc
     7201 agaagctgga tctctccaag atctacttca agaacgacaa gtccctgacc gatctctccc
     7261 agcaggtctt cgacgattac tccgtgatcg gcaccgcagt cctggaatac atcacccagc
     7321 agatcgcccc aaagaacctc gataacccat ccaagaagga acaggagctg atcgccaaga
     7381 agaccgaaaa ggctaagtac ctgtccctcg agaccatcaa gctggctctc gaagagttca
     7441 acaagcaccg cgacatcgat aagcagtgcc gcttcgaaga gatcctcgca aacttcgctg
     7501 caatcccaat gatcttcgac gaaatcgcac agaacaagga taacctggcc cagatctcca
     7561 tcaagtacca gaaccagggc aagaaggatc tgctccaggc ctccgctgag gacgatgtga
     7621 aggcaatcaa ggacctgctc gatcagacca acaacctgct ccacaagctg aagatcttcc
     7681 acatctccca gtccgaagac aaggccaaca tcctcgacaa ggatgagcac ttctacctgg
     7741 tgttcgaaga gtgctacttc gaactcgcta acatcgtccc actgtacaac aagatccgca
     7801 actacatcac ccagaagcca tactccgatg aaaagttcaa gctcaacttc gagaactcca
     7861 ccctggcaaa cggctgggac aagaacaagg aaccagataa caccgccatc ctcttcatca
     7921 aggacgataa gtactacctg ggcgtgatga acaagaagaa caacaagatc ttcgacgata
     7981 aggccatcaa ggaaaacaag ggcgagggct acaagaagat cgtgtacaag ctgctcccag
     8041 gcgctaacaa gatgctccca aaggtcttct tctccgcaaa gtccatcaag ttctacaacc
     8101 catccgaaga tatcctgcgc atccgcaacc actccaccca caccaagaac ggctccccac
     8161 agaagggcta cgaaaagttc gagttcaaca tcgaagactg ccgcaagttc atcgatttct
     8221 acaagcagtc catctccaag cacccagagt ggaaggactt cggcttccgc ttctccgata
     8281 cccagcgcta caactccatc gatgaattct accgcgaagt ggagaaccag ggctacaagc
     8341 tgaccttcga aaacatctcc gagtcctaca tcgattccgt ggtcaaccag ggcaagctgt
     8401 acctcttcca gatctacaac aaggacttct ccgcttactc caagggccgc ccaaacctgc
     8461 acaccctcta ctggaaggca ctcttcgacg aacgcaacct gcaggatgtg gtctacaagc
     8521 tcaacggcga agcagagctg ttctaccgca agcagtccat cccaaagaag atcacccacc
     8581 cagccaagga agcaatcgcc aacaagaaca aggataaccc aaagaaggaa tccgtgttcg
     8641 agtacgacct gatcaaggat aagcgcttca ccgaggacaa gttcttcttc cactgcccaa
     8701 tcaccatcaa cttcaagtcc tccggcgcca acaagttcaa cgatgaaatc aacctgctcc
     8761 tgaaggagaa ggctaacgac gtgcacatcc tgtccatcga tcgcggcgaa cgccacctcg
     8821 cctactacac cctggtcgac ggcaagggca acatcatcaa gcaggacacc ttcaacatca
     8881 tcggcaacga tcgcatgaag accaactacc acgacaagct ggccgctatc gagaaggacc
     8941 gcgattccgc tcgcaaggat tggaagaaga tcaacaacat caaggaaatg aaggaaggct
     9001 acctctccca ggtggtccac gaaatcgcta agctggtgat cgagtacaac gcaatcgtgg
     9061 tcttcgaaga cctgaacttc ggcttcaagc gcggccgctt caaggtggag aagcaggtct
     9121 accagaagct ggaaaagatg ctcatcgaga agctgaacta cctcgtgttc aaggacaacg
     9181 aattcgataa gaccggcggc gtcctccgtg cataccagct gaccgcccca ttcgagacct
     9241 tcaagaagat gggcaagcag accggcatca tctactacgt gccagctggc ttcacctcta
     9301 agatctgccc agtgaccggc ttcgtcaacc agctctaccc aaagtacgaa tccgtctcca
     9361 agtcccagga gttcttctcc aagttcgaca agatctgcta caacctggat aagggctact
     9421 tcgaattctc cttcgactac aagaacttcg gcgataaggc agccaagggc aagtggacca
     9481 tcgcatcctt cggctcccgc ctcatcaact tccgcaactc cgacaagaac cacaactggg
     9541 atacccgcga agtgtaccca accaaggaac tggagaagct cctgaaggat tactccatcg
     9601 aatacggcca cggcgagtgc atcaaggctg caatctgcgg cgaatccgac aagaagttct
     9661 tcgcaaagct gacctctgtg ctcaacacca tcctgcagat gcgcaactcc aagaccggca
     9721 ccgagctgga ttacctcatc tccccagtgg ccgacgtcaa cggcaacttc ttcgattccc
     9781 gccaggctcc aaagaacatg ccacaggacg ctgatgcaaa cggcgcctac cacatcggtc
     9841 tgaagggtct catgctcctg ggtcgcatca agaacaacca ggaaggcaag aagctgaatc
     9901 tcgtcattaa gaacgaagaa tactttgaat ttgtccagaa ccgcaataac taagatcgac
     9961 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatgcc
    10021 atcatggccg cggtgatca
//
