LOCUS       sequence-320872-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning and constitutive expression of crRNAs in mycobacterium.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  AUTHORS   Yan MY, Yan HQ, Ren GX, Zhao JP, Guo XP, Sun YC
  TITLE     CRISPR-Cas12a-Assisted Recombineering in Bacteria.
  JOURNAL   Appl Environ Microbiol. 2017 Aug 17;83(17). pii: AEM.00947-17. doi: 
            10.1128/AEM.00947-17. Print 2017 Sep 1.
  PUBMED    28646112
REFERENCE   2  (bases 1 to 5621)
  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: "Appl 
            Environ Microbiol."; date: "2017-08-17"; volume: "83(17). pii: 
            AEM.00947-17. doi"; pages: " 10.1128/AEM.00947-17. Print 2017 Sep 1"
COMMENT     Sequence Label: pCR-Hyg
FEATURES             Location/Qualifiers
     source          1..5621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      579..625
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      4800..5388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5289..5308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctagacggt gaccacaacg cgcccgcttt gatcggggac gtctgcggcc gaccatttac
       61 gggtcttgtt gtcgttggcg gtcatgggcc gaacatactc acccggatcg gagggccgag
      121 gacaaggtcg aacgaggggc atgacccggt gcggggcttc ttgcactcgg cataggcgag
      181 tgctaagaat aacgttggca ctcgcgaccg gtgagtgcta ggtcgggacg gtgaggccag
      241 gcccgtcgtc gcagcgagtg gcagcgagga caacttgagc cgtccgtcgc gggcactgcg
      301 cccggccagc gtaagtagcg gggttgccgt cacccggtga cccccgtttc atccccgatc
      361 cggaggaatc acttcgcaat ggccaagaca attgcggatc cagctgcaga atttggcgcc
      421 agaggcactt cgaggctgat ttaggcaaaa acgggtctaa gaactttaaa taatttctac
      481 tgttgtagat tccgtctccg gcctccagta gtattcggcg aagcttgtct aagaacttta
      541 aataatttct actgttgtag atatcgactg ccaggcatca aataaaacga aaggctcagt
      601 cgaaagactg ggcctttcgt tttatgccat catggccgcg ggctagctta tcaccagccc
      661 gtcatcgtca acgcctgatc cgcggtgcgg acaggccgtg tcgcgaccgg ccgtgcggaa
      721 ttaagccggc ccgtaccctg tgaatagagg tccgctgtga cacaagaatc cctgttactt
      781 ctcgaccgta ttgattcgga tgattcctac gcgagcctgc ggaacgacca ggaattctgg
      841 gagccgctgg cccgccgagc cctggaggag ctcgggctgc cggtgccgcc ggtgctgcgg
      901 gtgcccggcg agagcaccaa ccccgtactg gtcggcgagc ccggctcggt gatcaagctg
      961 ttcggcgagc actggtgcgg tccggagagc ctcgcgtcgg agtcggaggc gtacgcggtc
     1021 ctggcggacg ccccggtgcc ggtgccccgc ctcctcggcc gcggcgagct gcggcccggc
     1081 accggagcct ggccgtggcc ctacctggtg atgagccgga tgaccggcac cacctggcgg
     1141 tccgcgatgg acggcacgac cgaccggaac gcgctgctcg ccctggcccg cgaactcggc
     1201 cgggtgctcg gccggctgca cagggtgccg ctgaccggga acaccgtgct caccccccat
     1261 tccgaggtct tcccggaact gctgcgggaa cgccgcgcgg cgaccgtcga ggaccaccgc
     1321 gggtggggct acctctcgcc ccggctgctg gaccgcctgg aggactggct gccggacgtg
     1381 gacacgctgc tggccggccg cgaaccccgg ttcgtccacg gcgacctgca cgggaccaac
     1441 atcttcgtgg acctggccgc gaccgaggtc accgggatcg tcgacttcac cgacgtctat
     1501 gcgggagact cccgctacag cctggtgcaa ctgcatctca acgccttccg gggcgaccgc
     1561 gagatcctgg ccgcgctgct cgacggggcg cagtggaagc ggaccgagga cttcgcccgc
     1621 gaactgctcg ccttcacctt cctgcacgac ttcgaggtgt tcgaggagac cccgctggat
     1681 ctctccggct tcaccgatcc ggaggaactg gcgcagttcc tctgggggcc gccggacacc
     1741 gcccccggag cctgacgccc cgggccgccc ggcgccgccc ccggcccccg gcggccatcg
     1801 acccaagtac cgccacctaa catacgttgc gtcgagagac gaaggcacca cgattaccgg
     1861 cgacgatgat ctattgccgg tctacaccga cgctggggtg attgcgtttg ctccgggaac
     1921 accgatcgct tcgttcaaaa tcggcggaag ataccccgtg cgtgcggtgc tgaaggaccg
     1981 caaagatgtt ccgtacggca ccgaggtctg caagtacggg atgaaatccg gcgagacctg
     2041 tgggccggtc gttgtggctg ccaagtacac gatcaccgct cagctgcgca ccgtacatgg
     2101 cgactcaggc gcgccgctgt accgcaagaa ccgcgacggc acagccgatt tgatcggcat
     2161 cgcttcgagc gttaacgagg acagtcgcac gacgaagttc ttctggatcg cgcccgtgct
     2221 ggaagcactc aacctcgaag cgtgtggttg cggagccatc tagcaaccac acgaaacatg
     2281 cgcaacgaac cgcgcaacga acaacgccta gaactggccc tagatgagct gactcgtatc
     2341 gttggtaaac ctagtttgac cagcatgttt taactacgtt cggtgagctg tcaacggggc
     2401 ctgtaacggc acaacgaacc gtgcaacgag agtggccacg gatgccacca caggcactac
     2461 aacggagttc gccacgtaca tcaccacaac caccgattct ggcggtgagc tccccgatat
     2521 tcagcggaaa tggcttggta tcgaccaaga ttcgtagaac cccgtctcgt ctggctggta
     2581 ttcaaaacgg gcgcaacgaa acacgcaacg agacaggcat ggcccaaacc agaaaactag
     2641 cgtctaccag gacttttacc tgtccgaccc gttgcaacgg aaccccccac ggaacccccg
     2701 cgacacccgc tccccaattg cgttagaaca gcggtggatt gtcggcttcg ttgtgggcct
     2761 tttgagccgc ttcctgttct gccgcacgct ctttcctcgc ccgatagccg agtcgcttaa
     2821 cggtgtccag atgcagccct aaatgtttgg ccgtttgcgg ccaagagtgg ccctcgtcgt
     2881 cgtgataggc gcggatgcgt tcgcggcgtg cagcctgctc ggcgagccac tcgctgcgtt
     2941 cctgcgccac gagccggacg atgtggcgtt cggatagtcc ggtgattcga gcgccttcgg
     3001 cggcggtcac gcgccgcttt ttgcggacag tcggctgccg gttgtagccg tcgctgtagc
     3061 cgtcgctcat agcaatgcct ccatggctga cgcggacttt gcgcgccgcg caactgtgct
     3121 cgccgccgtg cgcgctgctg cgcccttccg cgagatggcc gactggcgcg cactgagtgt
     3181 ggcctcgtag accacgatcc cgtccgccca aatgcgcgac ttggttgtga tccaacgcca
     3241 aatgctgttg gcgatggcgc ggacctcgct gtccggtagc ggtccgggac acacgtcgtt
     3301 gcacgagaat tcggcgtttc gcgtgtggca ctcggcatag atcgcgcggc cgagtccgtc
     3361 cacgttccgg gtcggcaggt agatccgcat gagggcggga cgataggccc acaacctgac
     3421 ggaatcgaac agtgcgcaat tccgccctag cggcgtcgga gccgctttgt acgtggtctg
     3481 ctgacgccag cgcggcggtg gcatgttcgc gccgagctcg gcctcgatgt ggctgagtgt
     3541 gtagagatct gagtggagcc attccgtttc ccaggcgatg tggccggggt ttttggtcat
     3601 gaggcctgag taactgcggt cgccgtcgac ggcgcgccga aggccttcgg cgcacgccgc
     3661 catgtatgcg agcggcttac gccgcgcgta ttcggtgcgt ggaacagggg cgttgagtgc
     3721 ccacactgcg tgtgcgtggc cgttggcgcg attgcccacg atcgcgttgg gcagcggatg
     3781 ggacccccgg gcgctgagcg ctcggagcgc tgcgtctgga tggtctacgt ccacgaccag
     3841 caggtttgcc agcgctgttg ggttcgcctc gatgtaccgg cggcctaggg ccgacgcgcg
     3901 gctttggcgg tagatcccct cgagcagatc gtcgcttgcc agcggccagt acggcagcca
     3961 gagctgctca aattcgtcgg cgacgtggct cacgcttggt agtagaccac gattaatcac
     4021 cggtgtatgg tccgacacga gctccaagtc agatatttcg ctgaggggcc accccacaac
     4081 tgcacactcc cccgctctcc cgtcgagccc tggtggtgga acaccagcga cagccgagca
     4141 cccccaacca cctgtaccaa ccaggaggaa cacatgcgtc gtttcgagga cgtttccggg
     4201 ccgctgagag ccgctgtggc ggccgtacac gccgccttag acccgttaga ccccatgccg
     4261 cctgaatgcg cgggtacgag ccacacagcg cccgaactta cggagctggt gggctcacct
     4321 ggctttatgg cgtacgaatc ggctgtgtgc gacctgttgg gcgaggtgag gtacgcgcta
     4381 ctcacgctgg caagggcgac agagccgccc caccgagccc gcacggccgc gcgcggtgtc
     4441 aacaaccggg tgagtcgtgc acaccagcag gtgttcgagg cttggctcga agtgcaggac
     4501 atcgtggcga acgccgcccg atgagccgcg ccttacgctg gctgccagcc gttcgcgggc
     4561 tggttggtgc agcgcgtcga gcggttagag gccctgcggt gttccaccac cgcagggcct
     4621 cgcccttttt aaggctgaat ttgcttgtct ccgaatccaa ctggcttgtc caagggtgta
     4681 tctacgctta atccaaagtt caaacgaggg gattacacat gactaacttc gataacgttc
     4741 tcggctcgat actcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4801 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4861 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4921 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4981 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5041 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5101 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5161 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5221 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5281 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5341 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5401 gcggcctttt tacggttcct ggccttttgc tggcattttg ctcacatgtt ctttcctgcg
     5461 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5521 cgcagccgaa cgaccgagcg cagcgagcgg ccctagttgg caatgggatt cacctttatg
     5581 ttgataagaa ataaaagaaa atgccaatag gatatcggca t
//
