LOCUS       sequence-73889-e        7635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kanamycin resistant; expresses luxAB-DAS+4 under the control of a 
            promoter containing PtetO-4C5G; integrates into the 
            mycobacteriophage att-L5 site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7635)
  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 7635)
  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-0X750-luxAB-DAS+4
FEATURES             Location/Qualifiers
     source          1..7635
                     /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    4986..5010
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5182..5206)
                     /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(7628..7635,1..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 tgaattcaag tttgtacaaa aaagcaggct gctaccaggc ctagatctgg ggaccctaga
     5041 ggtccccttt tttattttaa aaattttttc acgtgccagc cgattggtcg ctctgaccag
     5101 ggaaaatagc cctctgacct ggggatttgc gatccccgtc agtgacggag ataattgtga
     5161 gcgctcacaa ccgcatgcac cacccagctt tcttgtacaa agtggttaat taacaggatc
     5221 cccaaataag gaaatgttat gaaatttgga aacttccttc tcacttatca gccacctgag
     5281 ctatctcaga ccgaagtgat gaagcgattg gttaatctgg gcaaagcgtc tgaaggttgt
     5341 ggcttcgaca ccgtttggtt gctagagcac cacttcactg aatttgggtt gttagggaat
     5401 ccttatgttg ctgccgcaca cctattaggt gcgacagaaa cgctcaacgt tggcactgca
     5461 gctatcgtat tgccgactgc ccatccggtt cgacaagcag aagacgtaaa cctactggat
     5521 caaatgtcaa aaggacgatt ccgttttggt atttgtcgcg gtttgtacga taaagatttt
     5581 cgtgtctttg gtacagacat ggataacagc cgagccttaa tggactgttg gtatgacttg
     5641 atgaaagaag gcttcaatga aggctatatc gcggcggata acgaacatat taagttcccg
     5701 aaaatccaac tgaatccatc ggcttacaca caaggtggtg ctcctgttta tgtcgtcgcg
     5761 gagtcagcat caacgacaga atgggctgca gagcgtggcc taccaatgat tctaagctgg
     5821 atcatcaaca ctcacgagaa gaaagcgcag cttgatcttt acaacgaagt cgcgactgaa
     5881 catggctacg atgtgactaa gattgaccac tgtttgtctt acatcacctc cgtcgatcat
     5941 gactcaaata gagccaaaga tatttgccgc aacttcttgg gccattggta cgactcatac
     6001 gtgaatgcca ccaagatttt tgacgactct gaccaaacaa aaggttacga cttcaataaa
     6061 ggtcaatggc gtgattttgt gttgaaaggc cacaaagaca ccaatcgccg aattgattac
     6121 agctacgaaa tcaacccagt agggacgcct gaagagtgta tcgcgattat ccagcaagat
     6181 attgatgcga cgggtattga caatatttgt tgtggttttg aagcaaacgg ttctgaagaa
     6241 gaaattatcg catctatgaa gctattccag tctgatgtga tgccatatct caaagaaaaa
     6301 cagtaattaa tattttctaa aaggaaagag acatgaaatt tggattattc ttcctcaatt
     6361 ttatgaattc aaagcgttct tctgatcaag tcatcgaaga aatgttagat accgcacatt
     6421 acgtagatca gttgaagttt gacacgttgg ctgtttacga aaaccatttc tcgaacaatg
     6481 gtgtggttgg tgccccacta acagtggctg gttttttact tggtatgaca aagaacgcca
     6541 aagtggcttc gttgaatcac gtcattacca cgcatcatcc agtacgtgtg gcggaagaag
     6601 cgtgtctact tgaccaaatg agtgaaggcc gttttgcctt tggctttagt gattgtgaaa
     6661 agagtgcaga tatgcgcttc tttaatcgac caacggattc tcagtttcag ttgttcagtg
     6721 agtgtcacaa gatcatcaat gatgcattca ctactgggta ctgccatcca aacaatgatt
     6781 tttatagttt tcctaaaatc tccgttaacc cacacgcgtt cactgaaggc ggtcctgcgc
     6841 aatttgtgaa tgcgacgagc aaagaagtgg ttgaatgggc ggctaagtta gggcttccac
     6901 tcgtgtttag atgggacgac tcaaacgctc aaagaaaaga atacgccggt ttgtaccacg
     6961 aagttgctca ggcacatggt gtcgatgtta gtcaggttcg acacaagctg acgctgctgg
     7021 tcaaccaaaa tgtagatggt gaagcagcaa gggcagaagc tcgcgtgtat ttggaagagt
     7081 ttgtccgtga atcttactca aataccgact ttgagcaaaa aatgggagag ctgttgtcag
     7141 aaaatgccat cggtacttat gaagaaagta ctcaggcagc gcgagttgcg attgagtgtt
     7201 gtggtgccgc ggacctattg atgtcttttg agtcgatgga agataaagcg cagcaaagag
     7261 cggttatcga tgtggtaaac gccaacatcg tcaaatacca ctcggccgcc aacgacgaga
     7321 actactccga gaactacgcg gacgccagct aataactagt cgtttaactg atgctgaagg
     7381 ggcagcgatg ccccttatat caccattctt ttcgcgcgat agcgctaact aatagaggca
     7441 tttatatgga cgtactttca gcggttaagc aggaaaatat cgcagcgagc acagaaatcg
     7501 atgacttgat tttcatggga actcctcagc aatggtcatt gcaggaacaa aaacagccgg
     7561 atctcgagct cgcgaaagct ctgatttaaa tggatccccc gggctgcagg aattcgatat
     7621 caagcttcaa cttta
//
