LOCUS       sequence-321958-       10032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Helper plasmid pYC1640 for C-terminal tagging with degradation tag 
            ssrA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10032)
  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 10032)
  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: pYC1876
FEATURES             Location/Qualifiers
     source          1..10032
                     /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      3568..3595
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             4899..5273
                     /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"
     polyA_signal    5337..5471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5374..5393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5428..5447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6071..6090
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(9239..9827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9319..9338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccctatcag tgatagagat aatctgggag tccctatcag tgatagagaa ggcggtatcg
       61 atatgtcgga cctggtgctg ggcctggaca 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 aaaaccctgg
     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 gggcaagcaa 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 tgcgcgagag caagaccctg 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 ggcatcgaaa 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 acttcgccgc cgactcgcac cagcgcgact acgccctggc cgcctaaata ctctggccat
     3481 ttttctttac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3541 acgcgtgcgg ccgctgatta gctaagcaga aggccatcct gacggatggc ctttttctag
     3601 aaaaaaaaaa agcgccgcaa ctgcggcgct tttttttttt taattaagtg atagattgcg
     3661 atgcgtgtgc tcggttgata taatctgtca gatcaggagg acttcgcatg tcgcggttgg
     3721 ataagtccaa ggtcatcaac tcggcgctgg agctgctgaa cgaggtcggt atcgagggac
     3781 tgaccacgcg caagttggcc caaaagctcg gcgtggaaca accgaccctc tactggcacg
     3841 tgaagaacaa gcgcgcgctg ctggatgcgt tggccatcga gatgctggat cgccatcaca
     3901 cgcacttttg ccccctggag ggcgagagct ggcaggattt tctgcggaac aacgcgaagt
     3961 cgttccggtg cgccctcctg agccatcggg atggcgccaa agtccacttg ggcacccgcc
     4021 cgaccgagaa gcaatacgag accctggaga accagctggc cttcctctgt caacagggtt
     4081 tttcgttgga gaacgccctg tatgccctgt ccgccgtggg acacttcacc ttgggctgcg
     4141 tgttggagga tcaggaacat caggtggcga aagaggaacg ggagaccccc acgacggatt
     4201 cgatgccccc gttgttgcgc caggcgatcg aactgtttga ccaccagggc gccgaacccg
     4261 cctttttgtt cgggctggag ctgatcatct gcgggctgga aaagcaactg aagtgcgaaa
     4321 gcggttcgtg ataactagtg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
     4381 gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttggt gagagctttg
     4441 ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg
     4501 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc
     4561 cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     4621 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     4681 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     4741 ataattccga cccgcacgac cagcgttagc atgctcagta agttgagtgc atcaggctca
     4801 gctctgaatt gacagcacac cgccgtcgag gcaagcttga gcggggtgca ctcatcatag
     4861 tgcaggaaag aagctctaca tattcaggag gattcaccat ggccaagttg accagtgccg
     4921 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     4981 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     5041 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     5101 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     5161 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     5221 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgaccgacgc
     5281 cgaccaacac cgccggtccg acgcggcccg acgggtccga ggggggtcga cctcgaaact
     5341 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5401 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5461 atgtctggat catggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     5521 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     5581 tgacgactga atccggtgag aatggcaaaa gattatgcat ttctttccag acttgttcaa
     5641 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     5701 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     5761 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     5821 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     5881 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     5941 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     6001 tcagaaacaa ctctggcgca tggggcttcc catacaagcg atagattgtc gcacctgatt
     6061 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     6121 atcgcggcct cgagggataa ccgtaaaaaa aaaaagcccc gcaatcgcgg ggcttttttt
     6181 tttgctagcc ctgtgaagag ccattgataa tgctcttcat ccaaaaaaat aaaaaagggg
     6241 acctctaggg tccccaagtc gacaaaaaaa aaacaccctg ccataaaatg acagggtgtt
     6301 gatttcggca tgaagcctta tctttgtagc ttctttcgag tacaaaaaca tgtcttccta
     6361 atgaagacac tcccagatta tatctatcac tgatagggat cgcaaatccc caggtcagag
     6421 ggctattttc cctggtcaga ggatcccgcg gccgcgccac ggatgccacc acaggcacta
     6481 caacggagtt cgccacgtac atcaccacaa ccaccgattc tggcggtgag ctccccgata
     6541 ttcagcggaa atggcttggt atcgaccaag attcgtagaa ccccgtctcg tctggctggt
     6601 attcaaaacg ggcgcaacga aacacgcaac gagacaggca tggcccaaac cagaaaacta
     6661 gcgtctacca ggacttttac ctgtccgacc cgttgcaacg gaacccccca cggaaccccc
     6721 gcgacacccg ctccccaatt gcgctccgtg gcgtaatcgt cgccgtagcg ttctagccac
     6781 gcccgccagt gttcagccgc gctcatctgg ctaccgctct gcggtccttc atcggtcatg
     6841 aacgcattct atcagtagat cagtaactca gtagatacgc cctgcgggca gtttgtcgtg
     6901 cacggtgcac tcctcgtgcc gccagcggcg caccggcgtt gacggcggaa tcgcctcgtt
     6961 gggagtggac ggtgccatgc cttcgacgat agatcgttgc ggtgcgtctg aacacgcgct
     7021 cggagtgcgc tcagggcggg ggcgtcgagc gcggttggta agcacttggc ggatgtgttc
     7081 gcgggcggcg gcaatgcggg cctgcgctgc gtcggtgagc gtcgcccggg ccggcgtctg
     7141 gtcgatgcgg gtggcggcgt agccgccgcc ttttggcggt gctggcggcg tggtccacga
     7201 gagccgtcgt agccggctgg acagaaacgc tgcggggttg gtgatgtggt cgggccagct
     7261 gcatccgcgg gtgcgcatgt cggcgtcgag ggcgtcggtg atagctcgtg ccgaccagac
     7321 ggccggctcg atgcccgcgg cggtaagcgt gtcgcagatg gcgccgatgt ggccgcggcc
     7381 caggccgtgg gtgcgcgcga cgagctgggc ggcgagcaac tggatggcaa gtggtcgcgg
     7441 tgtggtgcgc cagcgccgcg gtggatcggg ttgttgttcg cggctatagg tggcgtgtgc
     7501 gcgtgcgcgc acgcttggtg acttactccc tacgggagtt aaaaaaccat cgccgcctgg
     7561 cggcggtagg tggaaatcgt gcacaggctg atcaccggcc ggctgggggg cgcgacgcgg
     7621 cacgaggtgg tacaccgagg gtcgccggcc cacgctgggg gtggttggcg agccgtggcc
     7681 gcgctgtgcc tccaccgccc agcgcgacac ccgcagcacc cgccatgccg cggtcacggt
     7741 gcgcacgtca catcccactg tgtcggcgat ggtggcgcga gtgaccgcca cgtggcggcc
     7801 ggtggcgtga tcggcgtgct cggccatcac cgccgcgatc gccaacaacg tggccgcggt
     7861 catcgacacc cgctgctccg cacacaactc gcccagcgcc ggcgactgcg cccaccgccg
     7921 cagaccgtcc aaccatgaca cgcgactcgt ccacatcggc gcgcgatgtt tcgcaccggc
     7981 gcgcagcacc cagtcacgcc gaccctgtgc cacccgggcg cgctcgacgt gcgggctgtg
     8041 tccggcatag gccgccgcgc cgagcggtgg cggtgaagaa cacgcccgat gactcggtcg
     8101 ggcggtgaga agatcacgat tcgcgcactt gctgcgtggc ttatctggta cgccgggcag
     8161 gcatgcgcta ccgtgagacg agtcgaccaa gacacgtccc tttcaggggc aacgggtgcg
     8221 gacccggacc tgagttggcg ctcaggtcga taccttcaga caaggcccct tcgggggcct
     8281 tcgtcatgtc agcgggcgaa tccgcacacc agctattgag atgtcacatc tgccaaggcc
     8341 cccgcagtcg agcagggcgg ggcctcagcg cgagctctca catcgccagc ggcagcacct
     8401 ccttgtcgag ttcgcggacg gcctcgggat gaccgactgc aatggcgagg aggtctgccg
     8461 agagctggct gacgctggtg ccgcgttggg cggccatttc gcgcagacgc gcgtacacga
     8521 ctgcggcagc gcggaccttg atctgctcgc gctgcccctt atgcggttgg gccatgccga
     8581 cgatcgtccc ccagagaaag tcctgtacct ggtaccgaca cgccgaagcg gtgctcggtt
     8641 ttcggccgag ttcaccccgg catgactgcc ggcgagacaa agaagtcaac gccaggcact
     8701 gcgcgaacac tttccatctg aaggcgttgt ctgacttggg agttgggggc gttcggcgcg
     8761 tgattttgtc ggtggtcccc aatacgctgg cccaatggag cacccctacc cgacgggacc
     8821 aactacccat ccggctccca actgcacact cccccgctct cccgtcgagc cctggtggtg
     8881 gaacaccagc gacagccgag cacccccaac cacctgtacc aaccaggagg aacacatgcg
     8941 tcgtttcgag gacgtttccg ggccgctgag agccgctgtg gcggccgtac acgccgcctt
     9001 agacccgtta gaccccctgc cgcctgaatg cgcgggtacg agccacacag cgcccgaact
     9061 tacggagctg gtgggctcac gcgttgcgct cggtcgttcg gctgcggcga gcggtatcag
     9121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     9181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     9241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     9301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     9361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctcccc tcgggaagcg
     9421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     9481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     9541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     9601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     9661 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     9721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     9781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     9841 tcttttctac ggggtctgac gctcatggaa ctagtggtct gacgctcatg gaacaataaa
     9901 cgaaaggctc agtcgaaaga ctgggccttt cgtttatctg ttgtttgaaa aaaaaaagcc
     9961 ccgcgattgc ggggcttttt tttttcatat gtctgaccag ggaaaatagc cctctgacct
    10021 ggggatttga ct
//
