LOCUS       pSBY1_FnCpf1cg               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
            //gt12365.cn
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
//