LOCUS       sequence-201575-        6043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR system used to genome editing in Mycobacterium smegmatis, 
            expresses crRNA to ligD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6043)
  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 6043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: pSBY2_ligD(FnCpf1)
FEATURES             Location/Qualifiers
     source          1..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(727..743)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(728..744)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     rep_origin      4818..5406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5307..5326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5560..5577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      5667..5694
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5814..5899
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5991..6018
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 attaagggag aacatgaagg caatcagtcg ggtgctgatc gcgatggttg cagccatcgc
       61 ggcgcttttc acgagcacag gcacctctca cgcaggcctg gacaacgagc tgagcctcgt
      121 tgatggccag gaccgcaccc tcaccgtgca gcagtgggac accttcctca atggtgtgtt
      181 ccccctggac cgcaaccgtc ttacccgtga gtggttccac tccggtcgcg ccaagtacat
      241 cgtggccggc cccggtgccg acgagttcga gggcacgctg gaactcggct accagatcgg
      301 cttcccgtgg tcgctgggtg tgggcatcaa cttcagctac accaccccga acatcctgat
      361 cgacgacggt gacatcatcg ctccgccgtt cggcctgaac tcggtcatca ccccgaacct
      421 gttccccggt gtgtcgatct cggcagatct gggcaacggc cccggcatcc aggaagtcgc
      481 aacgttctcg gtcgacgtct ccggcgccga gggtggcgtg gccgtgtcga acgcccacgg
      541 caccgtgacc ggtgcggccg gcggtgtgct gctgcgtccg ttcgcccgcc tgatcgcctc
      601 gaccggtgac tcggtcacca cctacggcga accctggaac atgaactgat tcctggaccg
      661 ccgttcgatt taaataagct ttagctaatt aattggggac cctagaggtc ccctttttta
      721 ttttatcgat accgtcgacc tcgagggggg gcccccgggg atcatcgagc cgagaacgtt
      781 atcgaagttg gtcatgtgta atcccctcgt ttgaactttg gattaagcgt agatacaccc
      841 ttggacaagc cagttggatt cggagacaag caaattcagc cttaaaaagg gcgaggccct
      901 gcggtggtgg aacaccgcag ggcctctaac cgctcgacgc gctgcaccaa ccagcccgcg
      961 aacggctggc agccagcgta aggcgcggct catcgggcgg cgttcgccac gatgtcctgc
     1021 acttcgagcc aagcctcgaa cacctgctgg tgtgcacgac tcacccggtt gttgacaccg
     1081 cgcgcggccg tgcgggctcg gtggggcggc tctgtcgccc ttgccagcgt gagtagcgcg
     1141 tacctcacct cgcccaacag gtcgcacaca gccgattcgt acgccataaa gccaggtgag
     1201 cccaccagct ccgtaagttc gggcgctgtg tggctcgtac ccgcgcattc aggcggcagg
     1261 gggtctaacg ggtctaaggc ggcgtgtacg gccgccacag cggctctcag cggcccggaa
     1321 acgtcctcga aacgacgcat gtgttcctcc tggttggtac aggtggttgg gggtgctcgg
     1381 ctgtcgctgg tgttccacca ccagggctcg acgggagagc gggggagtgt gcagttgtgg
     1441 ggtggcccct cagcgaaata tctgacttgg agctcgtgtc ggaccataca ccggtgatta
     1501 atcgtggtct actaccaagc gtgagccacg tcgccgacga atttgagcag ctctggctgc
     1561 cgtactggcc gctggcaagc gacgatctgc tcgaggggat ctaccgccaa agccgcgcgt
     1621 cggccctagg ccgccggtac atcgaggcga acccaacagc gctggcaaac ctgctggtcg
     1681 tggacgtaga ccatccagac gcagcgctcc gagcgctcag cgcccggggg tcccatccgc
     1741 tgcccaacgc gatcgtgggc aatcgcgcca acggccacgc acacgcagtg tgggcactca
     1801 acgcccctgt tccacgcacc gaatacgcgc ggcgtaagcc gctcgcatac atggcggcgt
     1861 gcgccgaagg ccttcggcgc gccgtcgacg gcgaccgcag ttactcaggc ctcatgacca
     1921 aaaaccccgg ccacatcgcc tgggaaacgg aatggctcca ctcagatctc tacacactca
     1981 gccacatcga ggccgagctc ggcgcgaaca tgccaccgcc gcgctggcgt cagcagacca
     2041 cgtacaaagc ggctccgacg ccgctagggc ggaattgcgc actgttcgat tccgtcaggt
     2101 tgtgggccta tcgtcccgcc ctcatgcgga tctacctgcc gacccggaac gtggacggac
     2161 tcggccgcgc gatctatgcc gagtgccacg cgcgaaacgc cgaattcccg tgcaacgacg
     2221 tgtgtcccgg accgctaccg gacagcgagg tccgcgccat cgccaacagc atttggcgtt
     2281 ggatcacaac caagtcgcgc atttgggcgg acgggatcgt ggtctacgag gccacactca
     2341 gtgcgcgcca gtcggccatc tcgcggaagg gcgcagcagc gcgcacggcg gcgagcacag
     2401 ttgcgcggcg cgcaaagtcc gcgtcagcca tggaggcatt gctatgagcg acggctacag
     2461 cgacggctac aaccggcagc cgactgtccg caaaaagcgg cgcgtgaccg ccgccgaagg
     2521 cgctcgaatc accggactat ccgaacgcca cgtcgtccgg ctcgtggcgc aggaacgcag
     2581 cgagtggctc gccgagcagg ctgcacgccg cgaacgcatc cgcgcctatc acgacgacga
     2641 gggccactct tggccgcaaa cggccaaaca tttcgggctg catctggaca ccgttaagcg
     2701 actcggctat cgggcgagga aagagcgtgc ggcagaacag gaagcggctc aaaaggccca
     2761 caacgaagcc gacaatccac cgctgttcta acgcaattgg ggagcgggtg tcgcgggggt
     2821 tccgtggggg gttccgttgc aacgggtcgg acaggtaaaa gtcctggtag acgctagttt
     2881 tctggtttgg gccatgcctg tctcgttgcg tgtttcgttg cgcccgtttt gaataccagc
     2941 cagacgagac ggggttctac gaatcttggt cgataccaag ccatttccgc tgaatatcgg
     3001 ggagctcacc gccagaatcg gtggttgtgg tgatgtacgt ggcgaactcc gttgtagtgc
     3061 ctgtggtggc atccgtggcc actctcgttg cacggttcgt tgtgccgtta caggccccgt
     3121 tgacagctca ccgaacgtag ttaaaacatg ctggtcaaac taggtttacc aacgatacga
     3181 gtcagctcat ctagggccag ttctaggcgt tgttcgttgc gcggttcgtt gcgcatgttt
     3241 cgtgtggttg ctagatggct ccgcaaccac acgcttcgag gttgagtgct tccagcacgg
     3301 gcgcgatcca gaagaacttc gtcgtgcgac tgtcctcgtt gatccttgcc gagctgggat
     3361 ggaagcccgg ccgacccacc ctggaggaga tgatcgagga tgccagggcc tttcacgccc
     3421 gccgctgctg agcgtccgcc gccgggcggc ccggggcgtc aggcgccggg ggcggtgtcc
     3481 ggcggccccc agaggaactg cgccagttcc tccggatcgg tgaagccgga gagatccagc
     3541 ggggtctcct cgaacacctc gaagtcgtgc aggaaggtga aggcgagcag ttcgcgggcg
     3601 aagtcctcgg tccgcttcca ctgcgccccg tcgagcagcg cggccaggat ctcgcggtcg
     3661 ccccggaagg cgttgagatg cagttgcacc aggctgtagc gggagtctcc cgcatagacg
     3721 tcggtgaagt cgacgatccc ggtgacctcg gtcgcggcca ggtccacgaa gatgttggtc
     3781 ccgtgcaggt cgccgtggac gaaccggggt tcgcggccgg ccagcagcgt gtccacgtcc
     3841 ggcagccagt cctccaggcg gtccagcagc cggggcgaga ggtagcccca cccgcggtgg
     3901 tcctcgacgg tcgccgcgcg gcgttcccgc agcagttccg ggaagacctc ggaatggggg
     3961 gtgagcacgg tgttcccggt cagcggcacc ctgtgcagcc ggccgagcac ccggccgagt
     4021 tcgcgggcca gggcgagcag cgcgttccgg tcggtcgtgc cgtccatcgc ggaccgccag
     4081 gtggtgccgg tcatccggct catcaccagg tagggccacg gccaggctcc ggtgccgggc
     4141 cgcagctcgc cgcggccgag gaggcggggc accggcaccg gggcgtccgc caggaccgcg
     4201 tacgcctccg actccgacgc gaggctctcc ggaccgcacc agtgctcgcc gaacagcttg
     4261 atcaccgggc cgggctcgcc gaccagtacg gggttggtgc tctcgccggg cacccgcagc
     4321 accggcggca ccggcagccc gagctcctcc agggctcggc gggccagcgg ctcccagaat
     4381 tcctggtcgt tccgcaggct cgcgtaggaa tcatccgaat caatacggtc gagaagtaac
     4441 agggattctt gtgtcacagc ggacctctat tcacagggta cgggccggct taattccgca
     4501 cggccggtcg cgacacggcc tgtccgcacc gcggatcagg cgttgacgat gacgggctgg
     4561 tcggccacgt cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4621 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4681 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4741 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4801 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4861 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4921 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4981 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5041 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5101 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5161 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5221 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5281 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5341 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5401 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5461 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5521 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5581 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5641 agctagaact agttgattag ctaagcagaa ggccatcctg acggatggcc tttttgcgtt
     5701 taatactgtt taaactctag actcgagtct agattgacag ctagctcagt cctaggtata
     5761 atggatccga atttctactg ttgtagatag cacgtcacac gcaacggtga tttaaataaa
     5821 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     5881 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     5941 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     6001 cctgacggat ggcctttttg cgtttctaca aactcttaag ctt
//
