LOCUS       sequence-73918-e        6920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kanamycin resistant; contains qtag28, expresses sspB under a 
            promoter containing PtetO, and constutively expresses single-chain 
            TetR10 (tsc10); integrates into the mycobacteriophage att-Tweety 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6920)
  AUTHORS   Kim JH, O'Brien KM, Sharma R, Boshoff HI, Rehren G, Chakraborty S, 
            Wallach JB, Monteleone M, Wilson DJ, Aldrich CC, Barry CE 3rd, Rhee 
            KY, Ehrt S, Schnappinger D
  TITLE     A genetic strategy to identify targets for the development of drugs 
            that prevent bacterial persistence.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Nov 4.
  PUBMED    24191058
REFERENCE   2  (bases 1 to 6920)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Nov 4."
COMMENT     Sequence Label: pGMCtKq28-TSC10M1-sspB
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..94
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     76..93
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(274..293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(275..293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(376..964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(456..475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1296..2111)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2019..2038
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2103..2122
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4344..4366
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4363..4379
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4363..4379
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4399..4419
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    5947..5971
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6360..6384)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(6913..6920,1..13))
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 ttatacatag ttgcttaagc tgcaggggac cctagaggtc ccctttttta ttttaaaaat
       61 tttttataac cggtgattta ggtgacacta tagaagtgga attcgatggc ctagctggca
      121 tcggtacgtg ccgcgcaacc atgtagtagt cctggagcgt gtccatctgg tgttcaagct
      181 tagcgactcc acgatccgac tagcaatgca tcctgtttgg catcggcagt gatggagtag
      241 cacccttggt gcatatggaa accgttgtgg tctccctata gtgagtcgta ttaatttacg
      301 cgtttcactg gccgacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      361 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      421 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      481 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      541 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
      601 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      661 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      721 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      781 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      841 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      901 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      961 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaact agtgaggtct
     1021 gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca
     1081 gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt
     1141 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
     1201 caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
     1261 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     1321 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1381 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1441 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1501 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaaga
     1561 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1621 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1681 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1741 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1801 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1861 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1921 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatg gggcttccca
     1981 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2041 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     2101 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2161 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtcgctt
     2221 tgttgctagc tggcgggaac ggcaatgctc ctggcggcgg cggggctggt gcccaggtct
     2281 ccgcacagag cggcggtgct ggcgctcgcg gccaggcgtg gttcttcgcg tactgacaag
     2341 aaacccccct ctttaggact cagtgtcctt gggagggggg ctttttgcgt tggcgcgcct
     2401 gcaggtcgag atttacagtc ggtcgcctac gcaaacgtcg atgggagcag ttcggacagc
     2461 ccctgcatgg cctccagatg cctcgcccga tccgcatgcg catagatccg ctgcgcatcc
     2521 acactcgcat gacccaagat ctccatacgc gtttgctcat ccacacccgc tgcgcgcagc
     2581 aatgtcgagg tggtgtgccg cgagttgtgc ggcggtaacg actcggttgg accgatcacc
     2641 ccagcagcgc ggaacacgcc acgccacacg tcgtagtccg aacggggatc gatcggcttc
     2701 ccctccttgt gccacaccaa gtcatgcgga ttgtcggtgc ggagtttctg catcgccaca
     2761 tacaacggcg gcaacaacgg aacctcacgc caaccagcgt ccgtcttcgg ccgggtgaac
     2821 aacaacgacc cctcacattc ctgatactcg aaatgcgccg gcaggtccca ccgggactgc
     2881 gggcatgccc atgcccgtgt cttcccgcaa ggccagtacg ggggtttctt gggcatacgg
     2941 tcgggccggg acagcggtga cggttcgggt agaggatccc cacagccgtg gacgcgggtt
     3001 tccgattgca actgccaagc gatggtgatc catccctgag cggggttgtc gacgtagggc
     3061 cagcgcaggc cgaggagttc cccacggcgg gcgcccgtca ggaaaccggc ggcgatccgc
     3121 accgcatccg gttcgtcgca cacctggaac gcggtgtgaa tgatgtgctg cgccacgtcc
     3181 gccgggaagc cgttgcgttt cttcttccgg tactcgggct tgtcgaccaa tgcggccaca
     3241 ttcctggtcg ccacaccctc cgctaccgca tcgtccaagg ctttctggac gatgacatgg
     3301 accagctcgg cggtgcggga ggccccgatc tcggagtgca ggtcccgcac atgctgcggg
     3361 gtgagtttgt cgatgcgttt cgcgccgagg atcgggttga tgtggttgtg gatggcggcc
     3421 cggtagtcgt tgaggacgcc ggggcggact ttacgtttgg cgtggatgtt gtcgatccag
     3481 tgcagcatcc acttctccac agttgtggat gaggtggtgg cgatgcggcc ctcttcgacg
     3541 tcgcggcgga gttgtttgag tttggccatg gcggtgttgc ggtcaacgga ggacacccat
     3601 ttgtagcggc ggttgccgtt gcggtctggg ggtagttcta ctcgtcccat ccatttgccg
     3661 tcggcgcgtt ggaagaacgc tccgtctccg cgggttctgc gtttcttagt tgccatcgtt
     3721 ttccctccca gggggtcacc ctacggttca ccctacggtg ctgcgcagca ttacgcagaa
     3781 ttgcgcagta tcgggggtct acctgcgggt ttgacaacgt ttctcctggt atgcagccta
     3841 tcaaccgctg actcttaatc agcgggtcgg gggttcgaaa ccctcacggc gcacaggtca
     3901 gaggccataa gcctcagagg ggatcaccct aaaggtaacc ctataggggt tttcactggt
     3961 cctacatgtc gtcgcgccgt cgtactctct tttcataggg gagcatgagg atccaactga
     4021 gcttgctgtg ccttcgattg agcggttgaa gcatctggag cgtcggcgga cagcgatcac
     4081 ggttcccaac gatggctgaa agaaggatga aatgactgca ctcatcgata agcttcgaat
     4141 tctgcagctg gatccatgga tatctagtgt taactattta attggggacc ctagaggtcc
     4201 ccttttttat tttaaaaatt ttttcnaagg ggacctctag ggtccccagc tggctagcga
     4261 gtcatgaggt naccatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4321 ccgcctttga gtgagctgat accagcggat aacaatttca cacaggaaac agctatgacc
     4381 atgattacgc caagctatca actttgtata gaaaagttgc agctggctag cgagtcatga
     4441 ggtcaccatg gtcattagga gccgctctcg cacttcagct gcttctccaa gccgcagatg
     4501 atcagctcca ggccgaacaa gaaggccggc tcggcgccct ggtggtcgaa cagctcgatg
     4561 gcctggcgca gcaacggcgg catgctgtcg gtggtcgggg tctcgcgctc ctccttggcg
     4621 acctggtgct cctggtcctc caacacgcag cccagggtga agtggcccac ggccgacagg
     4681 gcgtacaacg cgttctccag cgagaagccc tgctggcaca ggaaggccaa ctggttctcc
     4741 agggtctcgt actgcttctc ggtcggccgg gtgcccaggt gcaccttggc gccgtcgcgg
     4801 tggctcagca aggcgcagcg gaagctcttg gcgttgttgc gcaggaagtc ctgccagctc
     4861 tcgccctcca gcgggcagaa gtgggtgtgg tggcggtcca acatctcgat ggccagggcg
     4921 tcgagcaggg cccgcttgtt cttgacgtgc cagtacaggg tcggctgctc gacgcccagc
     4981 ttctgggcca gcttgcgggt ggtcaggccc tcgatgccga cctcgttcag cagctccagg
     5041 gcggagttga tgaccttgga cttgtccagg cgggatctag agccacctcc gcccgatccg
     5101 ccacctccag agcctccacc gccagaaccg cctccaccgg atccaccgcc tcctgagtcg
     5161 cccgactcac acttcaactg cttctccagc ccacaaatga tgagttccaa accgaacagg
     5221 aacgcgggct ccgcaccctg atgatcgaag agctcgatcg cctgccggag cagcgggggc
     5281 atcgaatccg tggtgggcgt ctcccgctct tccttcgcca cctgatgttc ttggtcttcg
     5341 aggacgcacc ccagcgtgaa atgcccgacc gcgctcagcg catacagggc gttctcgagg
     5401 gagaaccctt gctgacacaa gaacgcgagc tggttctcga gcgtttcata ctgcttctcc
     5461 gtgggccgcg taccgagatg gaccttcgcc ccgtcccgat gggagaggag cgcacaccgg
     5521 aaggacttcg cgttgttccg gaggaagtct tgccacgact ccccttccaa gggacagaag
     5581 tgcgtatgat gccggtcgag catttcgatc gcgagcgcat ccagcaacgc gcgcttgttc
     5641 ttcacatgcc aatacaacgt gggttgctcc acgcccaact tctgcgcgag cttccgggtc
     5701 gtcaaccctt cgatgcccac ctcattcaac agttccaacg cgctgttgat caccttgctc
     5761 ttatccaacc gcgacattgc gaagtgattc ctcctgatgg tactggtacc ggatccgtgg
     5821 caggagggtc gcctgcgtcg gcaactcgcc caggctacac gagagccctc gactgcgaaa
     5881 atgccgacca ggcaccaccg cactccaggc ttaacccggt gcgacgatcc aatattacga
     5941 gatctgacaa gtttgtacaa aaaagcaggc taccaggcct agatctgggg accctagagg
     6001 tccccttttt tattttaaaa attttttcac gtgccagccg attggactag aaatattgga
     6061 tcgtcggcac cgtcacggcc gtgggaggcg gcacgatccg cgacgtgatg atcggccgca
     6121 tccccacggt gctgcgcagt gagctctacg ccatcccggc gttgatctgt gcgttcgcac
     6181 gcacaggccc ggtgtgagaa gggtctctga cgagcgggag aactccctat cagtgataga
     6241 gtttgtcctc cctatcagtg atagataggc tctgggagta cccgtgtgta cgaccagcac
     6301 ggcatacatc atttcgacgc cgagagattc gccgcccgaa atgagcacga tccgcatgca
     6361 cccagctttc ttgtacaaag tgggctcgag gcatgcgaag gagatataca tatggacctg
     6421 tcgcagctga ccccgcgccg gccctacctg ctgcgcgcct tctacgagtg gttgctggac
     6481 aaccagctga ccccgcacct ggtggtggac gtgaccctgc cgggcgtgca ggtgccgatg
     6541 gagtacgcgc gcgacggcca gatcgtgctg aacatcgcgc cgcgcgccgt cggcaacctg
     6601 gagctggcga acgacgaggt gcgcttcaac gcgcgcttcg gtggcatccc gcggcaggtc
     6661 tcggtgccgc tggcggccgt gctggccatc tacgcccgcg agaacggcgc cggcaccatg
     6721 ttcgagccgg aggcggccta cgacgaggac acctcgatca tgaacgacga ggaggcctcg
     6781 gccgacaacg agaccgtgat gtcggtgatc gacggcgaca agccggacca cgacgatgac
     6841 acccacccgg acgatgagcc gccccagccg ccgcgcggcg gtcgcccggc cctgcgcgtc
     6901 gtgaagtaat agcaacttta
//
