LOCUS       sequence-320844-        8226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid expressing Sth1 Cas9 and the cognate sgRNA .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8226)
  AUTHORS   Yan MY, Li SS, Ding XY, Guo XP, Jin Q, Sun YC
  TITLE     A CRISPR-Assisted Nonhomologous End-Joining Strategy for Efficient 
            Genome Editing in Mycobacterium tuberculosis.
  JOURNAL   mBio. 2020 Jan 28;11(1). pii: mBio.02364-19. doi: 
            10.1128/mBio.02364-19.
  PUBMED    31992616
REFERENCE   2  (bases 1 to 8226)
  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: "mBio."; 
            date: "2020-01-28"; volume: "11(1). pii: mBio.02364-19. doi"; pages:
            " 10.1128/mBio.02364-19"
COMMENT     Sequence Label: pYC1446
FEATURES             Location/Qualifiers
     source          1..8226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3529..3556
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             6513..6887
                     /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      complement(7456..8044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7536..7555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccctatcag tgatagagat aatctgggag tccctatcag tgatagagaa ggcggtatcg
       61 atatgtcgga cctggtgctg ggcctggata tcggcatcgg ctcggtgggc gtgggcatcc
      121 tgaacaaggt gaccggcgag atcatccaca agaactcgcg catcttcccg gccgcccagg
      181 ccgagaacaa cctggtgcgc cgcaccaacc gccagggccg ccgcctggcc cgccgcaaga
      241 agcaccgccg cgtgcgcctg aaccgcctgt tcgaggagtc gggcctgatc accgacttca
      301 ccaagatctc gatcaacctg aacccgtacc agctgcgcgt gaagggcctg accgacgagc
      361 tgtcgaacga ggagctgttc atcgccctga agaacatggt gaagcaccgc ggcatctcgt
      421 acctggacga cgcctcggac gacggcaact cgtcggtggg cgactacgcc cagatcgtga
      481 aggagaactc gaagcagctg gagaccaaga ccccgggcca gatccagctg gagcgctacc
      541 agacctacgg ccagctgcgc ggcgacttca ccgtggagaa ggacggcaag aagcaccgcc
      601 tgatcaacgt gttcccgacc tcggcctacc gctcggaggc cctgcgcatc ctgcagaccc
      661 agcaggagtt caacccgcag atcaccgacg agttcatcaa ccgctacctg gagatcctga
      721 ccggcaagcg caagtactac cacggcccgg gcaacgagaa gtcgcgcacc gactacggcc
      781 gctaccgcac ctcgggcgag accctggaca acatcttcgg catcctgatc ggcaagtgca
      841 ccttctaccc ggacgagttc cgcgccgcca aggcctcgta caccgcccag gagttcaacc
      901 tgctgaacga cctgaacaac ctgaccgtgc cgaccgagac caagaagctg tcgaaggagc
      961 agaagaacca gatcatcaac tacgtgaaga acgagaaggc catgggcccg gccaagctgt
     1021 tcaagtacat cgccaagctg ctgtcgtgcg acgtggccga catcaagggc taccgcatcg
     1081 acaagtcggg caaggccgag atccacacct tcgaggccta ccgcaagatg aagaccctgg
     1141 agaccctgga catcgagcag atggaccgcg agaccctgga caagctggcc tacgtgctga
     1201 ccctgaacac cgagcgcgag ggcatccagg aggccctgga gcacgagttc gccgacggct
     1261 cgttctcgca gaagcaggtg gacgagctgg tgcagttccg caaggccaac tcgtcgatct
     1321 tcggcaaggg ctggcacaac ttctcggtga agctgatgat ggagctgatc ccggagctgt
     1381 acgagacctc ggaggagcag atgaccatcc tgacccgcct gggcaagcag aagaccacct
     1441 cgtcgtcgaa caagaccaag tacatcgacg agaagctgct gaccgaggag atctacaacc
     1501 cggtggtggc caagtcggtg cgccaggcca tcaagatcgt gaacgccgcc atcaaggagt
     1561 acggcgactt cgacaacatc gtgatcgaga tggcccgcga gaccaacgag gacgacgaga
     1621 agaaggccat ccagaagatc cagaaggcca acaaggacga gaaggacgcc gccatgctga
     1681 aggccgccaa ccagtacaac ggcaaggccg agctgccgca ctcggtgttc cacggccaca
     1741 agcagctggc caccaagatc cgcctgtggc accagcaggg cgagcgctgc ctgtacaccg
     1801 gcaagaccat ctcgatccac gacctgatca acaactcgaa ccagttcgag gtggaccaca
     1861 tcctgccgct gtcgatcacc ttcgacgact cgctggccaa caaggtgctg gtgtacgcca
     1921 ccgccaacca ggagaagggc cagcgcaccc cgtaccaggc cctggactcg atggacgacg
     1981 cctggtcgtt ccgcgagctg aaggccttcg tgcgcgagtc gaagaccctg tcgaacaaga
     2041 agaaggagta cctgctgacc gaggaggaca tctcgaagtt cgacgtgcgc aagaagttca
     2101 tcgagcgcaa cctggtggac acccgctacg cctcgcgcgt ggtgctgaac gccctgcagg
     2161 agcacttccg cgcccacaag atcgacacca aggtgtcggt ggtgcgcggc cagttcacct
     2221 cgcagctgcg ccgccactgg ggcatcgaga agacccgcga cacctaccac caccacgccg
     2281 tggacgccct gatcatcgcc gcctcgtcgc agctgaacct gtggaagaag cagaagaaca
     2341 ccctggtgtc gtactcggag gaccagctgc tggacatcga gaccggcgag ctgatctcgg
     2401 acgacgagta caaggagtcg gtgttcaagg ccccgtacca gcacttcgtg gacaccctga
     2461 agtcgaagga gttcgaggac tcgatcctgt tctcgtacca ggtggactcg aagttcaacc
     2521 gcaagatctc ggacgccacc atctacgcca cccgccaggc caaggtgggc aaggacaagg
     2581 ccgacgagac ctacgtgctg ggcaagatca aggacatcta cacccaggac ggctacgacg
     2641 ccttcatgaa gatctacaag aaggacaagt cgaagttcct gatgtaccgc cacgacccgc
     2701 agaccttcga gaaggtgatc gagccgatcc tggagaacta cccgaacaag cagatcaacg
     2761 acaagggcaa ggaggtgccg tgcaacccgt tcctgaagta caaggaggag cacggctaca
     2821 tccgcaagta ctcgaagaag ggcaacggcc cggagatcaa gtcgctgaag tactacgact
     2881 cgaagctggg caaccacatc gacatcaccc cgaaggactc gaacaacaag gtggtgctgc
     2941 agtcggtgtc gccgtggcgc gccgacgtgt acttcaacaa gaccaccggc aagtacgaga
     3001 tcctgggcct gaagtacgcc gacctgcagt tcgacaaggg caccggcacc tacaagatct
     3061 cgcaggagaa gtacaacgac atcaagaaga aggagggcgt ggactcggac tcggagttca
     3121 agttcaccct gtacaagaac gacctgctgc tggtgaagga caccgagacc aaggagcagc
     3181 agctgttccg cttcctgtcg cgcaccatgc cgaagcagaa gcactacgtg gagctgaagc
     3241 cgtacgacaa gcagaagttc gagggcggcg aggccctgat caaggtgctg ggcaacgtgg
     3301 ccaactcggg ccagtgcaag aagggcctgg gcaagtcgaa catctcgatc tacaaggtgc
     3361 gcaccgacgt gctgggcaac cagcacatca tcaagaacga gggcgacaag ccgaagctgg
     3421 acttctaaat actctggcca tttttcttta ccgcctttga gtgagctgat accgctcgcc
     3481 gcagccgaac gaccgagcgc aacgcgtgcg gccgctgatt agctaagcag aaggccatcc
     3541 tgacggatgg cctttttcta gaaaaaaaaa aagcgccgca actgcggcgc tttttttttt
     3601 ttaattaagt gatagattgc gatgcgtgtg ctcggttgat ataatctgtc agatcaggag
     3661 gacttcgcat gtcgcggttg gataagtcca aggtcatcaa ctcggcgctg gagctgctga
     3721 acgaggtcgg tatcgaggga ctgaccacgc gcaagttggc ccaaaagctc ggcgtggaac
     3781 aaccgaccct ctactggcac gtgaagaaca agcgcgcgct gctggatgcg ttggccatcg
     3841 agatgctgga tcgccatcac acgcactttt gccccctgga gggcgagagc tggcaggatt
     3901 ttctgcggaa caacgcgaag tcgttccggt gcgccctcct gagccatcgg gatggcgcca
     3961 aagtccactt gggcacccgc ccgaccgaga agcaatacga gaccctggag aaccagctgg
     4021 ccttcctctg tcaacagggt ttttcgttgg agaacgccct gtatgccctg tccgccgtgg
     4081 gacacttcac cttgggctgc gtgttggagg atcaggaaca tcaggtggcg aaagaggaac
     4141 gggagacccc cacgacggat tcgatgcccc cgttgttgcg ccaggcgatc gaactgtttg
     4201 accaccaggg cgccgaaccc gcctttttgt tcgggctgga gctgatcatc tgcgggctgg
     4261 aaaagcaact gaagtgcgaa agcggttcgt gataacgccg aaaaccccgc ttcggcgggg
     4321 ttttgccgcg gtctgcgttg tcgggaagat ttaaaataaa aaaggggacc tctagggtcc
     4381 ccaattaaat agttaacact agtattaaaa aaaaaagccc cgcgattgcg gggctttttt
     4441 ttttcgacgg cgagggttcc gaccgctgca actcccggtg caaccttgtc ccggtctatt
     4501 ctcttcactg caccagctcc aatctggtgt gaatgcccct cgtctgttcg cgcaggcggg
     4561 gggctctatt cgtttgtcag catcgaaagt agccagatca gggatgcgtt gcaaccgcgt
     4621 atgcccaggt cagaagagtc gcacaagagt tgcagacccc tggaaagaaa aatggccaga
     4681 gggcgaaaac accctctgac cagcggagcg ggcgacggga atcgaacccg cgtagctagt
     4741 ttggaagaat gggtgtctgc cgaccacata tatgggccgg tcaagatagg tttttacccc
     4801 ctctcggctg catcctctaa gtggaaagaa attgcaggtc gtagaagcgc gttgaagcct
     4861 gagagttgca caggagttgc aacccggtag ccttgttcac gacgagagga gacctagttg
     4921 gcacgtcgcg gatggggatc gctgaagact cagcgcagcg ggaggatcca agcctcatac
     4981 gtcaacccgc aggacggtgt gaggtactac gcgctgcaga cctacgacaa caagatggac
     5041 gccgaagcct ggctcgcggg cgagaagcgg ctcatcgaga tggagacctg gacccctcca
     5101 caggaccggg cgaagaaggc agccgccagc gccatcacgc tggaggagta cacccggaag
     5161 tggctcgtgg agcgcgacct cgcagacggc accagggatc tgtacagcgg gcacgcggag
     5221 cgccgcatct acccggtgct aggtgaagtg gcggtcacag agatgacgcc agctctggtg
     5281 cgtgcgtggt gggccgggat gggtaggaag cacccgactg cccgccggca tgcctacaac
     5341 gtcctccggg cggtgatgaa cacagcggtc gaggacaagc tgatcgcaga gaacccgtgc
     5401 cggatcgagc agaaggcagc cgatgagcgc gacgtagagg cgctgacgcc tgaggagctg
     5461 gacatcgtcg ccgctgagat cttcgtgcac taccggatcg cggcatacat cctggcgtgg
     5521 acgagcctcc ggttcggaga gctgatcgag cttcgccgca aggacatcgt ggacgacggc
     5581 atgacgatga agctccgggt gcgccgtggc gcttcccgcg tggggaacaa gatcgtcgtt
     5641 ggcaacgcca agaccgtccg gtcgaagcgt cctgtgacgg ttccgcctca cgtcgcggag
     5701 atgatccgag cgcacatgaa ggaccgtacg aagatgaaca agggccccga ggcattcctg
     5761 gtgaccacga cgcagggcaa ccggctgtcg aagtccgcgt tcaccaagtc gctgaagcgt
     5821 ggctacgcca agatcggtcg gccggaactc cgcatccacg acctccgcgc tgtcggcgct
     5881 acgttcgccg ctcaggcagg tgcgacgacc aaggagctga tggcccgcct cggtcacacg
     5941 actcctagga tggcgatgaa gtaccagatg gcgtctgagg cccgcgacga ggctatcgct
     6001 gaggcgatgt ccaagctggc caagacctcc tgaaacgcaa aaagcccccc tcccaaggac
     6061 actgagtcct aaagaggggg gtttcttgtc agtacgcgaa gaaccacgcc tggccgcgag
     6121 cgccagcacc gccgctctgt gcggagacct gggcaccagc cccgccgccg ccaggagcat
     6181 tgccgttccc gccagctagc ctctgccagt gttacaacca attaaccaat tctgattaga
     6241 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     6301 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccataatt
     6361 ccgacccgca cgaccagcgt tagcatgctc agtaagttga gtgcatcagg ctcagctctg
     6421 aattgacagc acaccgccgt cgaggcaagc ttgagcgggg tgcactcatc atagtgcagg
     6481 aaagaagctc tacatattca ggaggattca ccatggccaa gttgaccagt gccgttccgg
     6541 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     6601 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     6661 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     6721 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     6781 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     6841 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgaccg acgccgacca
     6901 acaccgccgg tccgacgcgg cccgacgggt ccgagggggg tcgacctcga aacttgtatc
     6961 aataaacgaa aggctcagtc gaaagactgg gcctttcgtt tatctgttgt ttgaaaaaaa
     7021 aaagcgccgc aactgcggcg cttttttttt tgaattctct gaccagggaa aatagccctc
     7081 tgacctgggg atttgcgatc cctatcagtg atagatataa tctgggagga gacgattaat
     7141 gcgtctcggt ttttgtactc gaaagaagct acaaagataa ggcttcatgc cgaaatcaac
     7201 accctgtcat tttatggcag ggtgtttttt ttttgtcgac ttggggaccc tagaggtccc
     7261 cttttttatt tttttggatg aagagcatta tcaatggctc ttcacaggaa gcttaaaaaa
     7321 aaaagccccg cgattgcggg gctttttttt tttaatacgg ttatccacag aatcagggga
     7381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7621 tctcccctcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7861 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8041 tcaagaagat cctttgatct tttctacggg gtctgacgct catggaacaa taaacgaaag
     8101 gctcagtcga aagactgggc ctttcgttta tctgttgttt gaaaaaaaaa agccccgcga
     8161 ttgcggggct tttttttttc atatgtctga ccagggaaaa tagccctctg acctggggat
     8221 ttgact
//
