LOCUS       sequence-73914-e        5805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kanamycin resistant; contains qtag20 and 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 5805)
  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 5805)
  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: pGMCKq20-T38S38-0X
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(37..83)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     340..362
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     359..375
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     359..375
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    395..415
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1600..1624
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1744..1768)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1774..1794)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1857..1875
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1857..1874
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2062..2081)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2063..2081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2164..2752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2244..2263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3084..3899)
                     /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     3807..3826
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3891..3910
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgatcaccgc ggccatgatg gccggacaaa caacagataa aacgaaaggc ccagtctttc
       61 gactgagcct ttcgttttat ttgatgcctg gcagtcgatc gtacgctagt taactacgtc
      121 gacatcgatt taattaatat gcatcaattg atttaaatac tagtgttaac tatttaattg
      181 gggaccctag aggtcccctt ttttatttta aaaatttttt cnaaggggac ctctagggtc
      241 cccagctggc tagcgagtca tgaggtnacc atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgatacca gcggataaca atttcacaca
      361 ggaaacagct atgaccatga ttacgccaag ctatcaactt tgtatagaaa agttgcagct
      421 ggctagcgag tcatgaggtc accatggtca ttaggagccg ctctcgcact tcagctgctt
      481 ctccaagccg cagatgatca gctccaggcc gaacaagaag gccggctcgg cgccctggtg
      541 gtcgaacagc tcgatggcct ggcgcagcaa cggcggcatg ctgtcggtgg tcggggtctc
      601 gcgctcctcc ttggcggccg tgtgctcctg gtcctccaac acgcagccca gggtgaagtg
      661 gcccacggcc gacagggcgt acaacgcgtt ctccagcgag aagccgttct cggtcatgaa
      721 gcgcagctgg gtctccacgg tgtcgtactg cttctcgtcc gggcgggtgc ccaggtgcac
      781 cttggcgccg tcgcggtagc gcagcagcgc gcggcggaac gacatggcgt tgttgcgcag
      841 gaacgactgc cacgactcgc cggccgccgg cagcgagtag tcgtggtggc gcgccaggat
      901 ctccaccgcc agggcgtcca gcagggcccg cttgttcttg acgtgccagt acagggtctt
      961 ctgcgcgacg cccagcttct gggccagctt gcgggtggtc acgccctcga tgccgacctc
     1021 gttgcccagg gccagggcgg agttgatgac cttggacttg tccaggcggg acattgcgaa
     1081 gtgattcctc ctgatggtac cctaggtcag atcaacggcc tggtcgacaa gggcaatacg
     1141 gtgatcgtca tcgaacataa cctggacgtg atcggccgtc aactcgttga ggtaccggtc
     1201 ggattcaaat ggttcgtcga cggtctgatc ccgcggtgtg caccgatgcc ctcacggttc
     1261 cagcggtcga ccaggtcctc gccgtgacgc cccttgatcg caaacgccac cgaatgcacc
     1321 acggccacag ccaaagccga cacgctggtt accacgacga tgttcacggg gggccctcgc
     1381 ggatctggcc caactgctgg acttccaccg aagcaatcga gccgatgtca cgctgcaact
     1441 ggtgcgggtg ggcgacccgc gggcattcgg ctgcgtaccc accgacgagg aggaccgcgt
     1501 agtcgccttt ctggagaaga cggagcagac ggtgctgttg ggcgcctgct ggattcccag
     1561 gtcttcgaac accgacacgc tcgcggcgtg atccaatatt caagtttgta caaaaaagca
     1621 ggctcgaata ccgggatgcg acagcgctgg gcccaggaga tccaccggtc cagggcctgt
     1681 ttaccttcct cgcccttgac cgaaaacaca tgccgcaggc tctctttgag caggtaggcg
     1741 cgaacccagc tttcttgtac aaagtggacg cgtcaacttt attatacata gttgcttaag
     1801 ctgcagggga ccctagaggt cccctttttt attttaaaaa ttttttataa ccggtgattt
     1861 aggtgacact atagaagtgg aattcgatgg cctagctggc atcggtacgt gccgcgcaac
     1921 catgtagtag tcctggagcg tgtccatctg gtgttcaagc ttgcaacttc ttcaagaggc
     1981 tgccgtcccg cgccgatacc atgctatagt atggcgttct tgctgctgat ggagcatgcc
     2041 atatggaaac cgttgtggtc tccctatagt gagtcgtatt aatttacgcg tttcactggc
     2101 cgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2161 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2281 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2341 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2401 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2461 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2521 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2581 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2641 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2701 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2761 tttgatcttt tctacggggt ctgacgctca gtggaactag tgaggtctgc ctcgtgaaga
     2821 aggtgttgct gactcatacc aggcctgaat cgccccatca tccagccaga aagtgaggga
     2881 gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt
     2941 tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa
     3001 agttcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt
     3061 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     3121 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     3181 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     3241 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     3301 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagatt atgcatttct
     3361 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     3421 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     3481 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     3541 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     3601 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     3661 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     3721 ctacctttgc catgtttcag aaacaactct ggcgcatggg gcttcccata caagcgatag
     3781 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     3841 tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata
     3901 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     3961 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtcgctttg ttgctagctg
     4021 gcgggaacgg caatgctcct ggcggcggcg gggctggtgc ccaggtctcc gcacagagcg
     4081 gcggtgctgg cgctcgcggc caggcgtggt tcttcgcgta ctgacaagaa acccccctct
     4141 ttaggactca gtgtccttgg gaggggggct ttttgcgttt caggaggtct tggccagctt
     4201 ggacatcgcc tcagcgatag cctcgtcgcg ggcctcagac gccatctggt acttcatcgc
     4261 catcctagga gtcgtgtgac cgagacgggc catcagctcc ttggtcgtcg cacctgcctg
     4321 agcggcgaac gtagcgccga cagcgcggag gtcgtggatg cggagttccg gccgaccgat
     4381 cttggcgtag ccacgcttca gcgacttggt gaacgcggac ttcgacagcc ggttgccctg
     4441 cgtcgtggtc accaggaatg cctcggggcc cttgttcatc ttcgtacggt ccttcatgtg
     4501 cgctcggatc atctccgcga cgtgaggcgg aaccgtcaca ggacgcttcg accggacggt
     4561 cttggcgttg ccaacgacga tcttgttccc cacgcgggaa gcgccacggc gcacccggag
     4621 cttcatcgtc atgccgtcgt ccacgatgtc cttgcggcga agctcgatca gctctccgaa
     4681 ccggaggctc gtccacgcca ggatgtatgc cgcgatccgg tagtgctcga agatctcagc
     4741 ggcgacgatg tccagctcct caggcgtcag cgcctctacg tcgcgctcat cggctgcctt
     4801 ctgctcgatc cggcacgggt tctctgcgat cagcttgtcc tcgaccgctg tgttcatcac
     4861 cgcccggagg acgttgtagg catgccggcg ggcagtcggg tgcttcctac ccatcccggc
     4921 ccaccacgca cgcaccagag ctggcgtcat ctctgtgacc gccacttcac ctagcaccgg
     4981 gtagatgcgg cgctccgcgt gcccgctgta cagatccctg gtgccgtctg cgaggtcgcg
     5041 ctccacgagc cacttccggg tgtactcctc cagcgtgatg gcgctggcgg ctgccttctt
     5101 cgcccggtcc tgtggagggg tccaggtctc catctcgatg agccgcttct cgcccgcgag
     5161 ccaggcttcg gcgtccatct tgttgtcgta ggtctgcagc gcgtagtacc tcacaccgtc
     5221 ctgcgggttg acgtatgagg cttggatcct cccgctgcgc tgagtcttca gcgatcccca
     5281 tccgcgacgt gccaactagg tctcctctcg tcgtgaacaa ggctaccggg ttgcaactcc
     5341 tgtgcaactc tcaggcttca acgcgcttct acgacctgca atttctttcc acttagagga
     5401 tgcagccgag agggggtaaa aacctatctt gaccggccca tatatgtggt cggcagacac
     5461 ccattcttcc aaactagcta cgcgggttcg attcccgtcg cccgctccgc tggtcagagg
     5521 gtgttttcgc cctctggcca tttttctttc caggggtctg caactcttgt gcgactcttc
     5581 tgacctgggc atacgcggtt gcaacgcatc cctgatctgg ctactttcga tgctgacaaa
     5641 cgaatagagc cccccgcctg cgcgaacaga cgaggggcat tcacaccaga ttggagctgg
     5701 tgcagtgaag agaatagacc gggacaaggt tgcaccggga gttgcagcgg tcggaaccct
     5761 cgccgtcggc gggctggcgt tcgccctgtc gttcacggct ctagc
//
