LOCUS       sequence-73896-e        8809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kanamycin resistant; expresses luxAB-DAS+4 under the control of a 
            promoter containing PtetO-4C5G; constitutively expresses reverse 
            TetR (tetR38); integrates into the mycobacteriophage att-L5 site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8809)
  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 8809)
  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: pGMCK-T38S38-750-luxAB-DAS+4
FEATURES             Location/Qualifiers
     source          1..8809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(21..37)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(30..52)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    596..730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(633..652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     687..706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(926..1514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1006..1025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1846..2661)
                     /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     2569..2588
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2653..2672
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(4604..4650)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4907..4929
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4926..4942
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4926..4942
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4962..4982
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    6167..6191
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6357..6381)
                     /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(8802..8809,1..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 ttgataattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
       61 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      121 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggggattg
      241 gtggcgacga ctcctggagc ccgtcagtat cggcggaatt ccagctgagc gccggtcgct
      301 accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca tgtctgcccg
      361 tatttcgcgt aaggaaatcc attatgtact atttaaaaaa cacaaacttt tggatgttcg
      421 gtttattctt tttcttttac ttttttatca tgggagccta cttcccgttt ttcccgattt
      481 ggctacatga catcaaccat atcagcaaaa gtgatacggg tattattttt gccgctattt
      541 ctctgttctc gctattattc caaccgctgt ttggtctgct ttctgacaaa ctcggaactt
      601 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      661 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      721 tgtctggatc ctctagagtc gacctgcagg tttaaacgcg gccgcacgcg ttgcgctcgg
      781 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      841 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      901 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      961 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1021 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1081 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     1141 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1201 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1261 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1321 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1381 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1441 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1501 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaact
     1561 agtgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat
     1621 catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt
     1681 tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga
     1741 tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt
     1801 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     1861 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     1921 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     1981 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2041 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2101 tggcaaaaga ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2161 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2221 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     2281 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     2341 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     2401 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     2461 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatg
     2521 gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     2581 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt
     2641 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     2701 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     2761 aacgtcgctt tgttgctagc tggcgggaac ggcaatgctc ctggcggcgg cggggctggt
     2821 gcccaggtct ccgcacagag cggcggtgct ggcgctcgcg gccaggcgtg gttcttcgcg
     2881 tactgacaag aaacccccct ctttaggact cagtgtcctt gggagggggg ctttttgcgt
     2941 ttcaggaggt cttggccagc ttggacatcg cctcagcgat agcctcgtcg cgggcctcag
     3001 acgccatctg gtacttcatc gccatcctag gagtcgtgtg accgagacgg gccatcagct
     3061 ccttggtcgt cgcacctgcc tgagcggcga acgtagcgcc gacagcgcgg aggtcgtgga
     3121 tgcggagttc cggccgaccg atcttggcgt agccacgctt cagcgacttg gtgaacgcgg
     3181 acttcgacag ccggttgccc tgcgtcgtgg tcaccaggaa tgcctcgggg cccttgttca
     3241 tcttcgtacg gtccttcatg tgcgctcgga tcatctccgc gacgtgaggc ggaaccgtca
     3301 caggacgctt cgaccggacg gtcttggcgt tgccaacgac gatcttgttc cccacgcggg
     3361 aagcgccacg gcgcacccgg agcttcatcg tcatgccgtc gtccacgatg tccttgcggc
     3421 gaagctcgat cagctctccg aaccggaggc tcgtccacgc caggatgtat gccgcgatcc
     3481 ggtagtgctc gaagatctca gcggcgacga tgtccagctc ctcaggcgtc agcgcctcta
     3541 cgtcgcgctc atcggctgcc ttctgctcga tccggcacgg gttctctgcg atcagcttgt
     3601 cctcgaccgc tgtgttcatc accgcccgga ggacgttgta ggcatgccgg cgggcagtcg
     3661 ggtgcttcct acccatcccg gcccaccacg cacgcaccag agctggcgtc atctctgtga
     3721 ccgccacttc acctagcacc gggtagatgc ggcgctccgc gtgcccgctg tacagatccc
     3781 tggtgccgtc tgcgaggtcg cgctccacga gccacttccg ggtgtactcc tccagcgtga
     3841 tggcgctggc ggctgccttc ttcgcccggt cctgtggagg ggtccaggtc tccatctcga
     3901 tgagccgctt ctcgcccgcg agccaggctt cggcgtccat cttgttgtcg taggtctgca
     3961 gcgcgtagta cctcacaccg tcctgcgggt tgacgtatga ggcttggatc ctcccgctgc
     4021 gctgagtctt cagcgatccc catccgcgac gtgccaacta ggtctcctct cgtcgtgaac
     4081 aaggctaccg ggttgcaact cctgtgcaac tctcaggctt caacgcgctt ctacgacctg
     4141 caatttcttt ccacttagag gatgcagccg agagggggta aaaacctatc ttgaccggcc
     4201 catatatgtg gtcggcagac acccattctt ccaaactagc tacgcgggtt cgattcccgt
     4261 cgcccgctcc gctggtcaga gggtgttttc gccctctggc catttttctt tccaggggtc
     4321 tgcaactctt gtgcgactct tctgacctgg gcatacgcgg ttgcaacgca tccctgatct
     4381 ggctactttc gatgctgaca aacgaataga gccccccgcc tgcgcgaaca gacgaggggc
     4441 attcacacca gattggagct ggtgcagtga agagaataga ccgggacaag gttgcaccgg
     4501 gagttgcagc ggtcggaacc ctcgccgtcg gcgggctggc gttcgccctg tcgttcacgg
     4561 ctctagctga tcaccgcggc catgatggcc ggacaaacaa cagataaaac gaaaggccca
     4621 gtctttcgac tgagcctttc gttttatttg atgcctggca gtcgatcgta cgctagttaa
     4681 ctacgtcgac atcgatttaa ttaatatgca tcaattgatt taaatactag tgttaactat
     4741 ttaattgggg accctagagg tccccttttt tattttaaaa attttttcna aggggacctc
     4801 tagggtcccc agctggctag cgagtcatga ggtnaccatg ttctttcctg cgttatcccc
     4861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccagcg gataacaatt
     4921 tcacacagga aacagctatg accatgatta cgccaagcta tcaactttgt atagaaaagt
     4981 tgcagctggc tagcgagtca tgaggtcacc atggtcatta ggagccgctc tcgcacttca
     5041 gctgcttctc caagccgcag atgatcagct ccaggccgaa caagaaggcc ggctcggcgc
     5101 cctggtggtc gaacagctcg atggcctggc gcagcaacgg cggcatgctg tcggtggtcg
     5161 gggtctcgcg ctcctccttg gcggccgtgt gctcctggtc ctccaacacg cagcccaggg
     5221 tgaagtggcc cacggccgac agggcgtaca acgcgttctc cagcgagaag ccgttctcgg
     5281 tcatgaagcg cagctgggtc tccacggtgt cgtactgctt ctcgtccggg cgggtgccca
     5341 ggtgcacctt ggcgccgtcg cggtagcgca gcagcgcgcg gcggaacgac atggcgttgt
     5401 tgcgcaggaa cgactgccac gactcgccgg ccgccggcag cgagtagtcg tggtggcgcg
     5461 ccaggatctc caccgccagg gcgtccagca gggcccgctt gttcttgacg tgccagtaca
     5521 gggtcttctg cgcgacgccc agcttctggg ccagcttgcg ggtggtcacg ccctcgatgc
     5581 cgacctcgtt gcccagggcc agggcggagt tgatgacctt ggacttgtcc aggcgggaca
     5641 ttgcgaagtg attcctcctg atggtaccct aggtcagatc aacggcctgg tcgacaaggg
     5701 caatacggtg atcgtcatcg aacataacct ggacgtgatc ggccgtcaac tcgttgaggt
     5761 accggtcgga ttcaaatggt tcgtcgacgg tctgatcccg cggtgtgcac cgatgccctc
     5821 acggttccag cggtcgacca ggtcctcgcc gtgacgcccc ttgatcgcaa acgccaccga
     5881 atgcaccacg gccacagcca aagccgacac gctggttacc acgacgatgt tcacgggggg
     5941 ccctcgcgga tctggcccaa ctgctggact tccaccgaag caatcgagcc gatgtcacgc
     6001 tgcaactggt gcgggtgggc gacccgcggg cattcggctg cgtacccacc gacgaggagg
     6061 accgcgtagt cgcctttctg gagaagacgg agcagacggt gctgttgggc gcctgctgga
     6121 ttcccaggtc ttcgaacacc gacacgctcg cggcgtgatc caatattcaa gtttgtacaa
     6181 aaaagcaggc taccaggcct agatctgggg accctagagg tccccttttt tattttaaaa
     6241 attttttcac gtgccagccg attggtcgct ctgaccaggg aaaatagccc tctgacctgg
     6301 ggatttgcga tccccgtcag tgacggagat aattgtgagc gctcacaacc gcatgcaccc
     6361 agctttcttg tacaaagtgg ttaattaaca ggatcccaaa taaggaaatg ttatgaaatt
     6421 tggaaacttc cttctcactt atcagccacc tgagctatct cagaccgaag tgatgaagcg
     6481 attggttaat ctgggcaaag cgtctgaagg ttgtggcttc gacaccgttt ggttgctaga
     6541 gcaccacttc actgaatttg ggttgttagg gaatccttat gttgctgccg cacacctatt
     6601 aggtgcgaca gaaacgctca acgttggcac tgcagctatc gtattgccga ctgcccatcc
     6661 ggttcgacaa gcagaagacg taaacctact ggatcaaatg tcaaaaggac gattccgttt
     6721 tggtatttgt cgcggtttgt acgataaaga ttttcgtgtc tttggtacag acatggataa
     6781 cagccgagcc ttaatggact gttggtatga cttgatgaaa gaaggcttca atgaaggcta
     6841 tatcgcggcg gataacgaac atattaagtt cccgaaaatc caactgaatc catcggctta
     6901 cacacaaggt ggtgctcctg tttatgtcgt cgcggagtca gcatcaacga cagaatgggc
     6961 tgcagagcgt ggcctaccaa tgattctaag ctggatcatc aacactcacg agaagaaagc
     7021 gcagcttgat ctttacaacg aagtcgcgac tgaacatggc tacgatgtga ctaagattga
     7081 ccactgtttg tcttacatca cctccgtcga tcatgactca aatagagcca aagatatttg
     7141 ccgcaacttc ttgggccatt ggtacgactc atacgtgaat gccaccaaga tttttgacga
     7201 ctctgaccaa acaaaaggtt acgacttcaa taaaggtcaa tggcgtgatt ttgtgttgaa
     7261 aggccacaaa gacaccaatc gccgaattga ttacagctac gaaatcaacc cagtagggac
     7321 gcctgaagag tgtatcgcga ttatccagca agatattgat gcgacgggta ttgacaatat
     7381 ttgttgtggt tttgaagcaa acggttctga agaagaaatt atcgcatcta tgaagctatt
     7441 ccagtctgat gtgatgccat atctcaaaga aaaacagtaa ttaatatttt ctaaaaggaa
     7501 agagacatga aatttggatt attcttcctc aattttatga attcaaagcg ttcttctgat
     7561 caagtcatcg aagaaatgtt agataccgca cattacgtag atcagttgaa gtttgacacg
     7621 ttggctgttt acgaaaacca tttctcgaac aatggtgtgg ttggtgcccc actaacagtg
     7681 gctggttttt tacttggtat gacaaagaac gccaaagtgg cttcgttgaa tcacgtcatt
     7741 accacgcatc atccagtacg tgtggcggaa gaagcgtgtc tacttgacca aatgagtgaa
     7801 ggccgttttg cctttggctt tagtgattgt gaaaagagtg cagatatgcg cttctttaat
     7861 cgaccaacgg attctcagtt tcagttgttc agtgagtgtc acaagatcat caatgatgca
     7921 ttcactactg ggtactgcca tccaaacaat gatttttata gttttcctaa aatctccgtt
     7981 aacccacacg cgttcactga aggcggtcct gcgcaatttg tgaatgcgac gagcaaagaa
     8041 gtggttgaat gggcggctaa gttagggctt ccactcgtgt ttagatggga cgactcaaac
     8101 gctcaaagaa aagaatacgc cggtttgtac cacgaagttg ctcaggcaca tggtgtcgat
     8161 gttagtcagg ttcgacacaa gctgacgctg ctggtcaacc aaaatgtaga tggtgaagca
     8221 gcaagggcag aagctcgcgt gtatttggaa gagtttgtcc gtgaatctta ctcaaatacc
     8281 gactttgagc aaaaaatggg agagctgttg tcagaaaatg ccatcggtac ttatgaagaa
     8341 agtactcagg cagcgcgagt tgcgattgag tgttgtggtg ccgcggacct attgatgtct
     8401 tttgagtcga tggaagataa agcgcagcaa agagcggtta tcgatgtggt aaacgccaac
     8461 atcgtcaaat accactcggc cgccaacgac gagaactact ccgagaacta cgcggacgcc
     8521 agctaataac tagtcgttta actgatgctg aaggggcagc gatgcccctt atatcaccat
     8581 tcttttcgcg cgatagcgct aactaataga ggcatttata tggacgtact ttcagcggtt
     8641 aagcaggaaa atatcgcagc gagcacagaa atcgatgact tgattttcat gggaactcct
     8701 cagcaatggt cattgcagga acaaaaacag ccggatctcg agctcgcgaa agctctgatt
     8761 taaatggatc ccccgggctg caggaattcg atatcaagct tcaacttta
//
