LOCUS       sequence-12744-e        7871 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 7871)
  AUTHORS   van Kessel JC, Hatfull GF
  TITLE     Recombineering in Mycobacterium tuberculosis.
  JOURNAL   Nat Methods. 2007 Feb . 4(2):147-52.
  PUBMED    17179933
REFERENCE   2  (bases 1 to 7871)
  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: "Nat 
            Methods. 2007 Feb . 4(2):147-52."
COMMENT     Sequence Label: pJV52
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4739..4785
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4935..4954)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4946..5761
                     /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(5019..5038)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6093..6681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6582..6601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaccgcta ccggacagcg aggtccgcgc catcgccaac agcatttggc gttggatcac
       61 aaccaagtcg cgcatttggg cggacgggat cgtggtctac gaggccacac tcagtgcgcg
      121 ccatgcggcc atctcgcgga agggcgcagc agcgcgcacg gcggcgagca cagttgcgcg
      181 gcgcgcaaag tccgcgtcag ccatggaggc attgctatga gcgacggcta cagcgacggc
      241 tacagcgacg gctacaactg gcagccgact gtccgcaaaa agcggcgcgt gaccgccgcc
      301 gaaggcgctc gaatcaccgg actatccgaa cgccacgtcg tccggctcgt ggcgcaggaa
      361 cgcagcgagt ggttcgccga gcaggctgca cgccgcgaac gcatccgcgc ctatcacgac
      421 gacgagggcc actcttggcc gcaaacggcc aaacatttcg ggctgcatct ggacaccgtt
      481 aagcgactcg gctatcgggc gaggaaagag cgtgcggcag aacaggaagc ggctcaaaag
      541 gcccacaacg aagccgacaa tccaccgctg ttctaacgca attggggagc gggtgtcgcg
      601 ggggttccgt ggggggttcc gttgcaacgg gtcggacagg taaaagtcct ggtagacgct
      661 agttttctgg tttgggccat gcctgtctcg ttgcgtgttt cgttgcgtcc gttttgaata
      721 ccagccagac gagacggggt tctacgaatc ttggtcgata ccaagccatt tccgctgaat
      781 atcgtggagc tcaccgccag aatcggtggt tgtggtgatg tacgtggcga actccgttgt
      841 agtgcttgtg gtggcatccg tggcgcggcc gcggtaccag atctttaaat ctagataaag
      901 aagtgacgcg gtctcaagcg tcgagcgtcg tcagcgtgtc gaggatgtcg aagtcgtagc
      961 cgtcggcgct ggcgatgtag acctgctggt cgaattgact gtcgcgcata cacatcgggc
     1021 cccggggccc gtcgaacccg acatcgtgcg cggatgccat caggtccggt atctcggggg
     1081 agtgggcccg ctggaagatg gcctcgagcg caacgagacc ctcgtaacag gattcggcca
     1141 tcgcgttgag cggtggcgcg tcggcgccgt agcgggcgac gtagctgccc atcaggtcca
     1201 tggcacccgc ggtggccagt gaactgaagt acgccgcggc gacatagagg ttttcggtgg
     1261 agccggcgcc gctggccagc agcatgttct cctccatcag cgggctgaac cgcgccatgc
     1321 ggtcgtgccc gccggcgcgc gcgaactcgc ggttgaacaa cacggcgtcc tggccgacga
     1381 gcagcatcaa cacggcctgc gcccccgacg cgatggcctt gcggaccggt gcgcggaaat
     1441 cgtcggtgcc gtacgggacg tagatctccc gtctgagctc gaggtccaga tctcggcagt
     1501 acgcgcgggc ggccgcggcg gaacggcgcg gccagatgta gtcatcgccg accaggcacc
     1561 aggaccggat gccgaagtgg tcgcgcagcc aggcgagcgc gggcgcgatc tggatctgcg
     1621 gtgtctcgcc tgtgcagaac acgcccggtg tgcgttcacc gccctcgtac aacgaggtgt
     1681 agacgtacgg gatgcggtcg cggaccaccg gggagatgcg gttgcgcacg gccgagatgt
     1741 gccagccggt cacggcgtcg agaccgtgac ctcgcaaccg gtcggcgacg gtccgggcga
     1801 cgtcgtcgcc gggccgtccg ccgtcgagca cctcgatggt gaccttgcgg ccctgcaggc
     1861 cgcctcggtc gttgacctcc ttggccgcga gctcggccac ggcctcgcac gaaggcgcga
     1921 agattcccgc tggcccttga agcggaatca ccagcccgac gcggaactca acctcgccgt
     1981 cctgcactcc agatcaccgt cgatcccgtg tagtctgcgc ttcaaagctt tctagcagaa
     2041 ataattcatt ctgaacagac cccgccgtcg acacgaggag acacccacca tggccgccgg
     2101 acagcagcgc cgccccaacc tcctgctgcc gttggtgcgt ctgacccacc tcgcggagtc
     2161 ggcgatcgaa cgcgtgctcg cggactcgtc gctcaagatc gaggactggc gcgtgctcga
     2221 cgagttggcc ggacggcgca ccgtgcccat gagcgatctc gcgcaggcca cgctgatcac
     2281 gggtccgact ctcaccagaa ccgtcgatcg ccttgtgtcg caagggatca tctaccggac
     2341 tgccgatctg catgaccgcc ggcgggtgct cgtggcgttg accccgcggg ggcggacgct
     2401 gcgcaaccgc ctggtggacg cggtagccga ggccgagtgt gcggcttttg aatcgtgcgg
     2461 gctggacgtc gaccagttgc gcgaactcgt cgacaccacc tcgaatttga cttcgtaacc
     2521 acccgcgccc ggccggcgtt cacccttgac ttttattttc atctggatat atttcgggtg
     2581 aatggaaagg ggtgaccatg ccgacctaca cattccgttg ttcccactgc ggtcccttcg
     2641 atctcacctg cgcgatctcc gagcgcgatg cggcggcgac ctgtccggag tgccggacgc
     2701 cggcgcgccg ggtcttcggt tcggtagggc tgacgacatt caccgcggga catcaccgcg
     2761 cattcgacgc ggcgtccgcg agcgccgaaa gtcccacggt ggtgaagtcg attcccgcag
     2821 gcgcggaccg cccgcgggcc ccgcgccgca atcccggtct accgagtctg ccgaggtact
     2881 agcgacatgg gtggcgtcgg gctcttctac gtgggtgcgg tgctcatcat cgacgggctg
     2941 atgctgctgg gccgcatcag cccacgaggc gcaacaccgc tgaacttctt cgtcggcgga
     3001 ctgcaggtgg tgacgcctac ggtgctgatc ctgcagtccg gcggagacgc ggccgtgatc
     3061 ttcgcggcct ccgggctcta cctgttcggc ttcacctacc tgtgggtggc catcaacaac
     3121 gtgaccgact gggacggaga aggtctcgga tggttctcgc tgttcgtcgc gatcgccgca
     3181 ctcggctact cgtggcacgc gttcaccgcc gaggccgacc cggcgttcgg ggtgatctgg
     3241 ctgctgtggg cagtgctgtg gttcatgctg ttcctgctgc tcggcctggg gcacgacgca
     3301 ctggggcccg ccgtcgggtt cgtcgcggtg gccgaaggcg tgatcaccgc cgccgtgccg
     3361 gccttcctga tcgtgtcggg caactgggaa accggcccgc tccccgccgc ggtcatcgcc
     3421 gtgatcggtt ttgccgcagt tgttctcgca taccccatcg ggcgccgtct cgcagcgccg
     3481 tcagtcacca accctccacc ggccgcgctc gcggccacca cccgataaga gaaagggagt
     3541 ccacatatgt aacggatcca gctgcagaat tcgaagctta tcgatgtcga cgtagttaag
     3601 gggattacac atatggctga aaatgctgtc accaagcagg attcgcccaa ggcacccgag
     3661 acgatctcgc aagtgctgca ggtgctcgtg ccgcagctgg cgcgtgcagt gcccaagggt
     3721 atggaccccg accgcatcgc gcggatcgtg caaaccgaga tccgcaagtc gcgcaacgcg
     3781 aaagctgctg gcatcgctaa gcagtccctc gacgactgca cgcaagagtc atttgccggt
     3841 gcgctgctga cctcggccgc gctcggtctc gagcccggtg tcaacggcga gtgctacctc
     3901 gtgccctacc gcgacacccg gcgcggcgtg gtcgagtgcc agctgatcat cggctaccag
     3961 ggcatcgtca aactgttctg gcagcacccg cgcgcctccc ggatcgacgc gcagtgggtc
     4021 ggcgcgaacg acgaattcca ttacaccatg ggcctcaatc cgacgctgaa acacgtgaag
     4081 gccaagggtg atcggggtaa tccggtctac ttctacgcga tcgtcgaggt gaccggcgct
     4141 gagccgctgt gggacgtgtt caccgccgac gagatcaggg aattgcgtcg cggcaaggtc
     4201 ggatcctcgg gcgacatcaa ggacccgcag cgctggatgg agcggaagac cgcgctcaaa
     4261 caggtgctga agctggcacc gaagacgacg cggctcgacg cggcgatccg cgccgacgat
     4321 cgcccgggca ccgacctgtc acagtcgcag gcgctcgcgc tgccgtcgac cgtcaagccg
     4381 acggccgact acatcgacgg cgagatcgcc gagccgcacg aggtcgatac gccaccgaag
     4441 tcgtcgcgcg cacaacgcgc gcagcgcgcc accgcgccgg cgcccgacgt gcagatggcc
     4501 aatcccgatc agctgaagcg cctcggcgag atccagaagg ccgaaaagta caacgacgcc
     4561 gattggttca agttcctcgc cgattcggcc ggcgtcaaag ccacgcgcgc cgccgacctc
     4621 acgttcgacg aggcgaaggc tgtcatcgac atgttcgacg ggcccaacgc atgagcgccc
     4681 cggcgaattc cgacgcggtg gttgatctgc aactagcgta cgatcgactg ccaggcatca
     4741 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatgccat catggccgcg
     4801 gtgatcagct agccacctga cgtcgggggg gggggaaagc cacgttgtgt ctcaaaatct
     4861 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     4921 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctcgag
     4981 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     5041 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagccccatg cgccagagtt
     5101 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     5161 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     5221 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
     5281 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     5341 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     5401 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     5461 ctggcctgtt gaacaagtct ggaaagaaat gcataatctt ttgccattct caccggattc
     5521 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     5581 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     5641 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     5701 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     5761 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     5821 cggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc
     5881 gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac
     5941 aacaaagctc tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag
     6001 gcctggtatg agtcagcaac accttcttca cgaggcagac ctcactagtt ccactgagcg
     6061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6481 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcaacgcg
     6841 tgagcccacc agctccgtaa gttcgggtgc tgtgtggctc gtacccgcgc attcaggcgg
     6901 cagggggtct aacgggtcta aggcggcgtg tacggccgcc acagcggctc ttagcggccc
     6961 ggaaacgtcc tcgaaacgac gcatgtgttc ctcctggttg gtacaggtgg ttgggggtgc
     7021 tcggctgtcg ctggtgtttc atcatcaggg ctcgacggga gagcggggga gtgtgcagtt
     7081 gtggggtggc ccctcagcga aatatctgac ttggagctcg tgtcggacca tacaccggtg
     7141 attaatcgtg gtttattatc aagcgtgagc cacgtcgccg acgaatttga gcagctctgg
     7201 ctgccgtact ggtccctggc aagcgacgat ctgctcgagg ggatctaccg ccaaagccgc
     7261 gcgtcggccc taggccgccg gtacatcgag gcgaacccaa cagcgctggc aaacctgctg
     7321 gtcgtggacg tagaccatcc agacgcagcg ctccgagcgc tcagcgcccg ggggtcccat
     7381 ccgctgccca acgcgatcgt gggcaatcgc gccaacggcc acgcacacgc agtgtgggca
     7441 ctcaacgccc ctgttccacg caccgaatac gcgcggcgta agccgctcgc atacatggcg
     7501 gcgtgcgccg aaggccttcg gcgcgccgtc gatggcgacc gcagttactc aggcctcatg
     7561 accaaaaacc ccggccacat cgcctgggaa acggaatggc tccactcaga tctctacaca
     7621 ctcagccaca tcgaggccga gctcggcgcg aacatgccac cgccgcgctg gcgtcagcag
     7681 accacgtaca aagcggctcc gacgccgcta gggcggaatt gcgcactgtt cgattccgtc
     7741 aggttgtggg cctatcttcc cgccctcatg cggatctacc tgccgacccg gaacgtggac
     7801 ggactcggcc gcgcgatcta tgccgagtgc cacgcgcgaa acgccgaatt tccgtgcaac
     7861 gacgtgtgtc c
//
