LOCUS       sequence-142356-        7794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7794)
  AUTHORS   van Kessel JC, Hatfull GF
  TITLE     Efficient point mutagenesis in mycobacteria using single-stranded 
            DNA recombineering: characterization of antimycobacterial drug 
            targets.
  JOURNAL   Mol Microbiol. 2008 Mar . 67(5):1094-107.
  PUBMED    18221264
REFERENCE   2  (bases 1 to 7794)
  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: "Mol 
            Microbiol. 2008 Mar . 67(5):1094-107."
COMMENT     Sequence Label: pJV62
FEATURES             Location/Qualifiers
     source          1..7794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5700..5746
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5896..5915)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5907..6722
                     /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     complement(5980..5999)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7053..7641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7542..7561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgcgtgag cccaccagct ccgtaagttc gggcgctgtg tggctcgtac ccgcgcattc
       61 aggcggcagg gggtctaacg ggtctaaggc ggcgtgtacg gccgccacag cggctctcag
      121 cggcccggaa acgtcctcga aacgacgcat gtgttcctcc tggttggtac aggtggttgg
      181 gggtgctcgg ctgtcgctgg tgttccacca ccagggctcg acgggagagc gggggagtgt
      241 gcagttgtgg ggtggcccct cagcgaaata tctgacttgg agctcgtgtc ggaccataca
      301 ccggtgatta atcgtggtct actaccaagc gtgagccacg tcgccgacga atttgagcag
      361 ctctggctgc cgtactggcc gctggcaagc gacgatctgc tcgaggggat ctaccgccaa
      421 agccgcgcgt cggccctagg ccgccggtac atcgaggcga acccaacagc gctggcaaac
      481 ctgctggtcg tggacgtaga ccatccagac gcagcgctcc gagcgctcag cgcccggggg
      541 tcccatccgc tgcccaacgc gatcgtgggc aatcgcgcca acggccacgc acacgcagtg
      601 tgggcactca acgcccctgt tccacgcacc gaatacgcgc ggcgtaagcc gctcgcatac
      661 atggcggcgt gcgccgaagg ccttcggcgc gccgtcgatg gcgaccgcag ttactcaggc
      721 ctcatgacca aaaaccccgg ccacatcgcc tgggaaacgg aatggctcca ctcagatctc
      781 tacacactca gccacatcga ggccgagctc ggcgcgaaca tgccaccgcc gcgctggcgt
      841 cagcagacca cgtacaaagc ggctccgacg ccgctagggc ggaattgcgc actgttcgat
      901 tccgtcaggt tgtgggccta tcgtcccgcc ctcatgcgga tctacctgcc gacccggaac
      961 gtggacggac tcggccgcgc gatctatgcc gagtgccacg cgcgaaacgc cgaatttccg
     1021 tgcaacgacg tgtgtcccgg accgctaccg gacagcgagg tccgcgccat cgccaacagc
     1081 atttggcgtt ggatcacaac caagtcgcgc atttgggcgg acgggatcgt ggtctacgag
     1141 gccacactca gtgcgcgcca gtcggccatc tcgcggaagg gcgcagcagc gcgcacggcg
     1201 gcgagcacag ttgcgcggcg cgcaaagtcc gcgtcagcca tggaggcatt gctatgagcg
     1261 acggctacag cgacggctac aaccggcagc cgactgtccg caaaaagcgg cgcgtgaccg
     1321 ccgccgaagg cgctcgaatc accggactat ccgaacgcca cgtcgtccgg ctcgtggcgc
     1381 aggaacgcag cgagtggctc gccgagcagg ctgcacgccg cgaacgcatc cgcgcctatc
     1441 acgacgacga gggccactct tggccgcaaa cggccaaaca tttcgggctg catctggaca
     1501 ccgttaagcg actcggctat cgggcgagga aagagcgtgc ggcagaacag gaagcggctc
     1561 aaaaggccca caacgaagcc gacaatccac cgctgttcta acgcaattgg ggagcgggtg
     1621 tcgcgggggt tccgtggggg gttccgttgc aacgggtcgg acaggtaaaa gtcctggtag
     1681 acgctagttt tctggtttgg gccatgcctg tctcgttgcg tgtttcgttg cgcccgtttt
     1741 gaataccagc cagacgagac ggggttctac gaatcttggt cgataccaag ccatttccgc
     1801 tgaatatcgg ggagctcacc gccagaatcg gtggttgtgg tgatgtacgt ggcgaactcc
     1861 gttgtagtgc ctgtggtggc atccgtggcg cggccgcggt accagatctt taaatctaga
     1921 taaagaagtg acgcggtctc aagcgtcgag cgtcgccagc gtgtcgagga tgtcgaagtc
     1981 gtagccgtcg gcgctggcga tgtagacctg ctggtcgaat tgactgtcgc gcatacacat
     2041 cgggccccgg ggcccgtcga acccgacatc gtgcgcggat gccatcaggt ccggtatctc
     2101 gggggagtgg gcccgctgga agatggcctc gagcgcaagc agaccctcgt aacaggattc
     2161 ggccatcgcg ttgagcggtg gcgcgtcggc gccgtagcgg gcgacgtagc tgcccatcag
     2221 gtccatggca cccgcggtgg ccagtgaact gaagtacgcc gcggcgacat agaggttttc
     2281 ggtggagccg gcgccgctgg ccagcagcat gttctcctcc atcagcgggc tgaaccgcgc
     2341 catgcggtcg tgcccgccgg cgcgcgcgaa ctcgcggttg aacaacacgg cgtcctggcc
     2401 gacgagcagc atcaacacgg cctgcgcccc cgacgcgatg gccttgcgga caggtgcgcg
     2461 gaaatcgtcg gtgccgtacg ggacgtagat ctcccgtctg agctcgaggt ccagatctcg
     2521 gcagtacgcg cgggcggccg cggcggaacg gcgcggccag atgtagtcat cgccgaccag
     2581 gcaccaggac cggatgccga agtggtcgcg cagccaggcg agcgcgggcg cgatctggat
     2641 ctgcggtgtc tcgcctgtgc agaacacgcc cggtgtgcgt tcaccgccct cgtacaacga
     2701 ggtgtagacg tacgggatgc ggtcgcggac caccggggag atgcggttgc gcacggccga
     2761 gatgtgccag ccggtcacgg cgtcgagacc gtgacctcgc aaccggtcgg cgacggtccg
     2821 ggcgacgtcg tcgccgggcg ctccgccgtc gagcacctcg atggtgacct tgcggccctg
     2881 caggccgcct cggtcgttga cctccttggc cgcgagctcg gccacggcct cgcacgaagg
     2941 cgcgaagatt cccgctggcc cttgaagcgg aatcaccagc ccgacgcgga actcaacctc
     3001 gccgtcctgc actccagatc accgtcgatc ccgtgtagtc tgcgcttcaa agctttctag
     3061 cagaaataat tcattctgaa cagaccccgc cgtcgacacg aggagacacc caccatggcc
     3121 gccggacagc agcgccgccc caacctcctg ctgccgttgg tgcgtctgac ccacctcgcg
     3181 gagtcggcga tcgaacgcgt gctcgcggac tcgtcgctca agatcgagga ctggcgcgtg
     3241 ctcgacgagt tggccggacg gcgcaccgtg cccatgagcg atctcgcgca ggccacgctg
     3301 atcacgggtc cgactctcac cagaaccgtc gatcgccttg tgtcgcaagg gatcatctac
     3361 cggactgccg atctgcatga ccgccggcgg gtgctcgtgg cgttgacccc gcgggggcgg
     3421 acgctgcgca accgcctggt ggacgcggta gccgaggccg agtgtgcggc ttttgaatcg
     3481 tgcgggctgg acgtcgacca gttgcgcgaa ctcgtcgaca ccacctcgaa tttgacttcg
     3541 taaccacccg cgcccggcgc gggcgttcac ccttgacttt tattttcatc tggatatatt
     3601 tcgggtgaat ggaaaggggt gaccatgccg acctacacat tccgttgttc ccactgcggt
     3661 cccttcgatc tcacctgcgc gatctccgag cgcgatgcgg cggcgacctg tccggagtgc
     3721 cggacgccgg cgcgccgggt cttcggttcg gtagggctga cgacattcac cgcgggacat
     3781 caccgcgcat tcgacgcggc gtccgcgagc gccgaaagtc ccacggtggt gaagtcgatt
     3841 cccgcaggcg cggaccgccc gcgggccccg cgccgcaatc ccggtctacc gagtctgccg
     3901 aggtactagc gacatgggtg gcgtcgggct cttctacgtg ggtgcggtgc tcatcatcga
     3961 cgggctgatg ctgctgggcc gcatcagccc acgaggcgca acaccgctga acttcttcgt
     4021 cggcggactg caggtggtga cgcctacggt gctgatcctg cagtccggcg gagacgcggc
     4081 cgtgatcttc gcggcctccg ggctctacct gttcggcttc acctacctgt gggtggccat
     4141 caacaacgtg accgactggg acggagaagg tctcggatgg ttctcgctgt tcgtcgcgat
     4201 cgccgcactc ggctactcgt ggcacgcgtt caccgccgag gccgacccgg cgttcggggt
     4261 gatctggctg ctgtgggcag tgctgtggtt catgctgttc ctgctgctcg gcctggggca
     4321 cgacgcactg gggcccgccg tcgggttcgt cgcggtggcc gaaggcgtga tcaccgccgc
     4381 cgtgccggcc ttcctgatcg tgtcgggcaa ctgggaaacc ggcccgctcc ccgccgcggt
     4441 catcgccgtg atcggttttg ccgcagttgt tctcgcatac cccatcgggc gccgtctcgc
     4501 agcgccgtca gtcaccaacc ctccaccggc cgcgctcgcg gccaccaccc gataagagaa
     4561 agggagtcca catatggctg aaaatgctgt caccaagcag gattcgccca aggcacccga
     4621 gacgatctcg caagtgctgc aggtgctcgt gccgcagctg gcgcgtgcag tgcccaaggg
     4681 tatggacccc gaccgcatcg cgcggatcgt gcaaaccgag atccgcaagt cgcgcaacgc
     4741 gaaagctgct ggcatcgcta agcagtccct cgacgactgc acgcaagagt catttgccgg
     4801 tgcgctgctg acctcggccg cgctcggtct cgagcccggt gtcaacggcg agtgctacct
     4861 cgtgccctac cgcgacaccc ggcgcggcgt ggtcgagtgc cagctgatca tcggctacca
     4921 gggcatcgtc aaactgttct ggcagcaccc gcgcgcctcc cggatcgacg cgcagtgggt
     4981 cggcgcgaac gacgaattcc attacaccat gggcctcaat ccgacgctga aacacgtgaa
     5041 ggccaagggt gatcggggta atccggtcta cttctacgcg atcgtcgagg tgaccggcgc
     5101 tgagccgctg tgggacgtgt tcaccgccga cgagatcagg gaattgcgtc gcggcaaggt
     5161 cggatcctcg ggcgacatca aggacccgca gcgctggatg gagcggaaga ccgcgctcaa
     5221 acaggtgctg aagctggcac cgaagacgac gcggctcgac gcggcgatcc gcgccgacga
     5281 tcgcccgggc accgacctgt cacagtcgca ggcgctcgcg ctgccgtcga ccgtcaagcc
     5341 gacggccgac tacatcgacg gcgagatcgc cgagccgcac gaggtcgata cgccaccgaa
     5401 gtcgtcgcgc gcacaacgcg cgcagcgcgc caccgcgccg gcgcccgacg tgcagatggc
     5461 caatcccgat cagctgaagc gcctcggcga gatccagaag gccgaaaagt acaacgacgc
     5521 cgattggttc aagttcctcg ccgattcggc cggcgtcaaa gccacgcgcg ccgccgacct
     5581 cacgttcgac gaggcgaagg ctgtcatcga catgttcgac gggcccaacg catgagcgcc
     5641 ccggcgaatt ccgacgcggt ggttgatctg caactagcgt acgatcgact gccaggcatc
     5701 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatgcca tcatggccgc
     5761 ggtgatcagc tagccacctg acgtcggggg ggggggaaag ccacgttgtg tctcaaaatc
     5821 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     5881 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga
     5941 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     6001 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagccccat gcgccagagt
     6061 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     6121 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     6181 atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag
     6241 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     6301 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     6361 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     6421 gctggcctgt tgaacaagtc tggaaagaaa tgcataatct tttgccattc tcaccggatt
     6481 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     6541 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     6601 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     6661 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     6721 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     6781 gcggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc
     6841 cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta
     6901 caacaaagct ctcaccaacc gtggctccct cactttctgg ctggatgatg gggcgattca
     6961 ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctcactagt tccatgagcg
     7021 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     7201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7441 gagctatgag aaagcgccac gcttcccgag gggagaaagg cggacaggta tccggtaagc
     7501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgc
//
