LOCUS       sequence-320874-        5178 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 5178)
  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 5178)
  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-Zeo
FEATURES             Location/Qualifiers
     source          1..5178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      578..624
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             914..1288
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      4356..4944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4845..4864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagacggtg accacaacgc gcccgctttg atcggggacg tctgcggccg accatttacg
       61 ggtcttgttg tcgttggcgg tcatgggccg aacatactca cccggatcgg agggccgagg
      121 acaaggtcga acgaggggca tgacccggtg cggggcttct tgcactcggc ataggcgagt
      181 gctaagaata acgttggcac tcgcgaccgg tgagtgctag gtcgggacgg tgaggccagg
      241 cccgtcgtcg cagcgagtgg cagcgaggac aacttgagcc gtccgtcgcg ggcactgcgc
      301 ccggccagcg taagtagcgg ggttgccgtc acccggtgac ccccgtttca tccccgatcc
      361 ggaggaatca cttcgcaatg gccaagacaa ttgcggatcc agctgcagaa tttggcgcca
      421 gaggcacttc gaggctgatt taggcaaaaa cgggtctaag aactttaaat aatttctact
      481 gttgtagatt ccgtctccgg cctccagtag tattcggcga agcttgtcta agaactttaa
      541 ataatttcta ctgttgtaga tatcgactgc caggcatcaa ataaaacgaa aggctcagtc
      601 gaaagactgg gcctttcgtt ttatgccatc atggccgcgg gctagcttat caccagcccg
      661 tcatcgtcaa cgcctgatcc gcggtgcgga caggccgtgt cgcgaccggc cgtgcggaat
      721 taagccggcc cgtaccctgt gaatagaggt ccataggaat tccgacccgc acgaccagcg
      781 ttagcatgct cagtaagttg agtgcatcag gctcagctct gaattgacag cacaccgccg
      841 tcgaggcaat cttgagcggg gtgcactcat catagtgcag gaaagaagct ctacatattc
      901 aggaggattc accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
      961 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     1021 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     1081 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     1141 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     1201 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     1261 gcacttcgtg gccgaggagc aggactgacc gacgccgacc aacaccgccg gtccgacgcg
     1321 gcccgacggg tccgaggggg gtcgacctcg aaacttgtcc aagtaccgcc acctaacata
     1381 cgttgcgtcg agagacgaag gcaccacgat taccggcgac gatgatctat tgccggtcta
     1441 caccgacgct ggggtgattg cgtttgctcc gggaacaccg atcgcttcgt tcaaaatcgg
     1501 cggaagatac cccgtgcgtg cggtgctgaa ggaccgcaaa gatgttccgt acggcaccga
     1561 ggtctgcaag tacgggatga aatccggcga gacctgtggg ccggtcgttg tggctgccaa
     1621 gtacacgatc accgctcagc tgcgcaccgt acatggcgac tcaggcgcgc cgctgtaccg
     1681 caagaaccgc gacggcacag ccgatttgat cggcatcgct tcgagcgtta acgaggacag
     1741 tcgcacgacg aagttcttct ggatcgcgcc cgtgctggaa gcactcaacc tcgaagcgtg
     1801 tggttgcgga gccatctagc aaccacacga aacatgcgca acgaaccgcg caacgaacaa
     1861 cgcctagaac tggccctaga tgagctgact cgtatcgttg gtaaacctag tttgaccagc
     1921 atgttttaac tacgttcggt gagctgtcaa cggggcctgt aacggcacaa cgaaccgtgc
     1981 aacgagagtg gccacggatg ccaccacagg cactacaacg gagttcgcca cgtacatcac
     2041 cacaaccacc gattctggcg gtgagctccc cgatattcag cggaaatggc ttggtatcga
     2101 ccaagattcg tagaaccccg tctcgtctgg ctggtattca aaacgggcgc aacgaaacac
     2161 gcaacgagac aggcatggcc caaaccagaa aactagcgtc taccaggact tttacctgtc
     2221 cgacccgttg caacggaacc ccccacggaa cccccgcgac acccgctccc caattgcgtt
     2281 agaacagcgg tggattgtcg gcttcgttgt gggccttttg agccgcttcc tgttctgccg
     2341 cacgctcttt cctcgcccga tagccgagtc gcttaacggt gtccagatgc agccctaaat
     2401 gtttggccgt ttgcggccaa gagtggccct cgtcgtcgtg ataggcgcgg atgcgttcgc
     2461 ggcgtgcagc ctgctcggcg agccactcgc tgcgttcctg cgccacgagc cggacgatgt
     2521 ggcgttcgga tagtccggtg attcgagcgc cttcggcggc ggtcacgcgc cgctttttgc
     2581 ggacagtcgg ctgccggttg tagccgtcgc tgtagccgtc gctcatagca atgcctccat
     2641 ggctgacgcg gactttgcgc gccgcgcaac tgtgctcgcc gccgtgcgcg ctgctgcgcc
     2701 cttccgcgag atggccgact ggcgcgcact gagtgtggcc tcgtagacca cgatcccgtc
     2761 cgcccaaatg cgcgacttgg ttgtgatcca acgccaaatg ctgttggcga tggcgcggac
     2821 ctcgctgtcc ggtagcggtc cgggacacac gtcgttgcac gagaattcgg cgtttcgcgt
     2881 gtggcactcg gcatagatcg cgcggccgag tccgtccacg ttccgggtcg gcaggtagat
     2941 ccgcatgagg gcgggacgat aggcccacaa cctgacggaa tcgaacagtg cgcaattccg
     3001 ccctagcggc gtcggagccg ctttgtacgt ggtctgctga cgccagcgcg gcggtggcat
     3061 gttcgcgccg agctcggcct cgatgtggct gagtgtgtag agatctgagt ggagccattc
     3121 cgtttcccag gcgatgtggc cggggttttt ggtcatgagg cctgagtaac tgcggtcgcc
     3181 gtcgacggcg cgccgaaggc cttcggcgca cgccgccatg tatgcgagcg gcttacgccg
     3241 cgcgtattcg gtgcgtggaa caggggcgtt gagtgcccac actgcgtgtg cgtggccgtt
     3301 ggcgcgattg cccacgatcg cgttgggcag cggatgggac ccccgggcgc tgagcgctcg
     3361 gagcgctgcg tctggatggt ctacgtccac gaccagcagg tttgccagcg ctgttgggtt
     3421 cgcctcgatg taccggcggc ctagggccga cgcgcggctt tggcggtaga tcccctcgag
     3481 cagatcgtcg cttgccagcg gccagtacgg cagccagagc tgctcaaatt cgtcggcgac
     3541 gtggctcacg cttggtagta gaccacgatt aatcaccggt gtatggtccg acacgagctc
     3601 caagtcagat atttcgctga ggggccaccc cacaactgca cactcccccg ctctcccgtc
     3661 gagccctggt ggtggaacac cagcgacagc cgagcacccc caaccacctg taccaaccag
     3721 gaggaacaca tgcgtcgttt cgaggacgtt tccgggccgc tgagagccgc tgtggcggcc
     3781 gtacacgccg ccttagaccc gttagacccc atgccgcctg aatgcgcggg tacgagccac
     3841 acagcgcccg aacttacgga gctggtgggc tcacctggct ttatggcgta cgaatcggct
     3901 gtgtgcgacc tgttgggcga ggtgaggtac gcgctactca cgctggcaag ggcgacagag
     3961 ccgccccacc gagcccgcac ggccgcgcgc ggtgtcaaca accgggtgag tcgtgcacac
     4021 cagcaggtgt tcgaggcttg gctcgaagtg caggacatcg tggcgaacgc cgcccgatga
     4081 gccgcgcctt acgctggctg ccagccgttc gcgggctggt tggtgcagcg cgtcgagcgg
     4141 ttagaggccc tgcggtgttc caccaccgca gggcctcgcc ctttttaagg ctgaatttgc
     4201 ttgtctccga atccaactgg cttgtccaag ggtgtatcta cgcttaatcc aaagttcaaa
     4261 cgaggggatt acacatgact aacttcgata acgttctcgg ctcgatactc gttccactga
     4321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4501 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4741 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4981 ttttgctggc attttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5041 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5101 gagcggccct agttggcaat gggattcacc tttatgttga taagaaataa aagaaaatgc
     5161 caataggata tcggcatt
//
