LOCUS       sequence-209088-        8801 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 8801)
  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 8801)
  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: pJV53
FEATURES             Location/Qualifiers
     source          1..8801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5670..5716
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5866..5885)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5877..6692
                     /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(5950..5969)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7023..7611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7512..7531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaccgcta ccggacagcg aggtccgcgc catcgccaac agcatttggc gttggatcac
       61 aaccaagtcg cgcatttggg cggacgggat cgtggtctac gaggccacac tcagtgcgcg
      121 ccagtcggcc atctcgcgga agggcgcagc agcgcgcacg gcggcgagca cagttgcgcg
      181 gcgcgcaaag tccgcgtcag ccatggaggc attgctatga gcgacggcta cagcgacggc
      241 tacaaccggc agccgactgt ccgcaaaaag cggcgcgtga ccgccgccga aggcgctcga
      301 atcaccggac tatccgaacg ccacgtcgtc cggctcgtgg cgcaggaacg cagcgagtgg
      361 ctcgccgagc aggctgcacg ccgcgaacgc atccgcgcct atcacgacga cgagggccac
      421 tcttggccgc aaacggccaa acatttcggg ctgcatctgg acaccgttaa gcgactcggc
      481 tatcgggcga ggaaagagcg tgcggcagaa caggaagcgg ctcaaaaggc ccacaacgaa
      541 gccgacaatc caccgctgtt ctaacgcaat tggggagcgg gtgtcgcggg ggttccgtgg
      601 ggggttccgt tgcaacgggt cggacaggta aaagtcctgg tagacgctag ttttctggtt
      661 tgggccatgc ctgtctcgtt gcgtgtttcg ttgcgcccgt tttgaatacc agccagacga
      721 gacggggttc tacgaatctt ggtcgatacc aagccatttc cgctgaatat cggggagctc
      781 accgccagaa tcggtggttg tggtgatgta cgtggcgaac tccgttgtag tgcctgtggt
      841 ggcatccgtg gcgcggccgc ggtaccagat ctttaaatct agataaagaa gtgacgcggt
      901 ctcaagcgtc gagcgtcgcc agcgtgtcga ggatgtcgaa gtcgtagccg tcggcgctgg
      961 cgatgtagac ctgctggtcg aattgactgt cgcgcataca catcgggccc cggggcccgt
     1021 cgaacccgac atcgtgcgcg gatgccatca ggtccggtat ctcgggggag tgggcccgct
     1081 ggaagatggc ctcgagcgca agcagaccct cgtaacagga ttcggccatc gcgttgagcg
     1141 gtggcgcgtc ggcgccgtag cgggcgacgt agctgcccat caggtccatg gcacccgcgg
     1201 tggccagtga actgaagtac gccgcggcga catagaggtt ttcggtggag ccggcgccgc
     1261 tggccagcag catgttctcc tccatcagcg ggctgaaccg cgccatgcgg tcgtgcccgc
     1321 cggcgcgcgc gaactcgcgg ttgaacaaca cggcgtcctg gccgacgagc agcatcaaca
     1381 cggcctgcgc ccccgacgcg atggccttgc ggacaggtgc gcggaaatcg tcggtgccgt
     1441 acgggacgta gatctcccgt ctgagctcga ggtccagatc tcggcagtac gcgcgggcgg
     1501 ccgcggcgga acggcgcggc cagatgtagt catcgccgac caggcaccag gaccggatgc
     1561 cgaagtggtc gcgcagccag gcgagcgcgg gcgcgatctg gatctgcggt gtctcgcctg
     1621 tgcagaacac gcccggtgtg cgttcaccgc cctcgtacaa cgaggtgtag acgtacggga
     1681 tgcggtcgcg gaccaccggg gagatgcggt tgcgcacggc cgagatgtgc cagccggtca
     1741 cggcgtcgag accgtgacct cgcaaccggt cggcgacggt ccgggcgacg tcgtcgccgg
     1801 gcgctccgcc gtcgagcacc tcgatggtga ccttgcggcc ctgcaggccg cctcggtcgt
     1861 tgacctcctt ggccgcgagc tcggccacgg cctcgcacga aggcgcgaag attcccgctg
     1921 gcccttgaag cggaatcacc agcccgacgc ggaactcaac ctcgccgtcc tgcactccag
     1981 atcaccgtcg atcccgtgta gtctgcgctt caaagctttc tagcagaaat aattcattct
     2041 gaacagaccc cgccgtcgac acgaggagac acccaccatg gccgccggac agcagcgccg
     2101 ccccaacctc ctgctgccgt tggtgcgtct gacccacctc gcggagtcgg cgatcgaacg
     2161 cgtgctcgcg gactcgtcgc tcaagatcga ggactggcgc gtgctcgacg agttggccgg
     2221 acggcgcacc gtgcccatga gcgatctcgc gcaggccacg ctgatcacgg gtccgactct
     2281 caccagaacc gtcgatcgcc ttgtgtcgca agggatcatc taccggactg ccgatctgca
     2341 tgaccgccgg cgggtgctcg tggcgttgac cccgcggggg cggacgctgc gcaaccgcct
     2401 ggtggacgcg gtagccgagg ccgagtgtgc ggcttttgaa tcgtgcgggc tggacgtcga
     2461 ccagttgcgc gaactcgtcg acaccacctc gaatttgact tcgtaaccac ccgcgcccgg
     2521 cgcgggcgtt cacccttgac ttttattttc atctggatat atttcgggtg aatggaaagg
     2581 ggtgaccatg ccgacctaca cattccgttg ttcccactgc ggtcccttcg atctcacctg
     2641 cgcgatctcc gagcgcgatg cggcggcgac ctgtccggag tgccggacgc cggcgcgccg
     2701 ggtcttcggt tcggtagggc tgacgacatt caccgcggga catcaccgcg cattcgacgc
     2761 ggcgtccgcg agcgccgaaa gtcccacggt ggtgaagtcg attcccgcag gcgcggaccg
     2821 cccgcgggcc ccgcgccgca atcccggtct accgagtctg ccgaggtact agcgacatgg
     2881 gtggcgtcgg gctcttctac gtgggtgcgg tgctcatcat cgacgggctg atgctgctgg
     2941 gccgcatcag cccacgaggc gcaacaccgc tgaacttctt cgtcggcgga ctgcaggtgg
     3001 tgacgcctac ggtgctgatc ctgcagtccg gcggagacgc ggccgtgatc ttcgcggcct
     3061 ccgggctcta cctgttcggc ttcacctacc tgtgggtggc catcaacaac gtgaccgact
     3121 gggacggaga aggtctcgga tggttctcgc tgttcgtcgc gatcgccgca ctcggctact
     3181 cgtggcacgc gttcaccgcc gaggccgacc cggcgttcgg ggtgatctgg ctgctgtggg
     3241 cagtgctgtg gttcatgctg ttcctgctgc tcggcctggg gcacgacgca ctggggcccg
     3301 ccgtcgggtt cgtcgcggtg gccgaaggcg tgatcaccgc cgccgtgccg gccttcctga
     3361 tcgtgtcggg caactgggaa accggcccgc tccccgccgc ggtcatcgcc gtgatcggtt
     3421 ttgccgcagt tgttctcgca taccccatcg ggcgccgtct cgcagcgccg tcagtcacca
     3481 accctccacc ggccgcgctc gcggccacca cccgataaga gaaagggagt ccacatatgt
     3541 aacggatcca gctgcagaat tcgaagctta tcgatgtcga cgtagttacg agatcggcgg
     3601 ccgcatatga gtgtgcccac acaggacgga atgcaccggt tcgtcgacga ggacgtctac
     3661 cacgctgacc ggggctcgct gtcggtatcc ggcgcgaagc tgctgttgcc gccgtcgtgt
     3721 cccgcgaaat tccgctggga gatggacaac acccggaagc cgaaaaaggt ctgggacttc
     3781 ggacatgtcg cgcacaaact ggtgctcggc aagggtgccg agttcgagat cctcgacccc
     3841 gaggtgcacg ggctgaaggc ggacggtacg ccgtcggaga agccgaccgc gacgggcatg
     3901 tggcgcaagg ccgaggctga ggctcgcaaa cagggcaagg tgccgattca cgtcgacctg
     3961 ttcacgaagg cgtacgacat ggccgaaaag gtgcgtcagc acccgacagc cggcccgatc
     4021 ttcgccaatc ctgacggcga ggccgaggtc gcgctgtact acaccgaccc cgagaccggc
     4081 gtgcggctgc gtggccggat cgactggctc actgacgata tcgatgatta caagacgtcg
     4141 atgaccgcga acccggccga gctgaaaacc aagttctaca agctcggcta tttcatgcag
     4201 gcggcctggt acatcgatct actggtcgcc ctcgggctcg ccgagaaccc gcgattccgg
     4261 ttcatcacgc aggagaaaga accgccctac gtcgtgactc cgatccagta cgacgacgag
     4321 gcgatcgaag aggggcggcg ccgcaaccgc caggcgatcc ggctctacgc cgactgcatg
     4381 gaatcgggca agtggcctga ctacagcgac gacgtggtca cgatcagcct gccctcgtgg
     4441 gggctgccgc gaccgcagac cgtcggcgac gtcgtcaccg acagctatat ctacgacacc
     4501 gacccgctcg aagaggccga cccgattgaa ggggattaca tctatggctg aaaatgctgt
     4561 caccaagcag gattcgccca aggcacccga gacgatctcg caagtgctgc aggtgctcgt
     4621 gccgcagctg gcgcgtgcag tgcccaaggg tatggacccc gaccgcatcg cgcggatcgt
     4681 gcaaaccgag atccgcaagt cgcgcaacgc gaaagctgct ggcatcgcta agcagtccct
     4741 cgacgactgc acgcaagagt catttgccgg tgcgctgctg acctcggccg cgctcggtct
     4801 cgagcccggt gtcaacggcg agtgctacct cgtgccctac cgcgacaccc ggcgcggcgt
     4861 ggtcgagtgc cagctgatca tcggctacca gggcatcgtc aaactgttct ggcagcaccc
     4921 gcgcgcctcc cggatcgacg cgcagtgggt cggcgcgaac gacgaattcc attacaccat
     4981 gggcctcaat ccgacgctga aacacgtgaa ggccaagggt gatcggggta atccggtcta
     5041 cttctacgcg atcgtcgagg tgaccggcgc tgagccgctg tgggacgtgt tcaccgccga
     5101 cgagatcagg gaattgcgtc gcggcaaggt cggatcctcg ggcgacatca aggacccgca
     5161 gcgctggatg gagcggaaga ccgcgctcaa acaggtgctg aagctggcac cgaagacgac
     5221 gcggctcgac gcggcgatcc gcgccgacga tcgcccgggc accgacctgt cacagtcgca
     5281 ggcgctcgcg ctgccgtcga ccgtcaagcc gacggccgac tacatcgacg gcgagatcgc
     5341 cgagccgcac gaggtcgata cgccaccgaa gtcgtcgcgc gcacaacgcg cgcagcgcgc
     5401 caccgcgccg gcgcccgacg tgcagatggc caatcccgat cagctgaagc gcctcggcga
     5461 gatccagaag gccgaaaagt acaacgacgc cgattggttc aagttcctcg ccgattcggc
     5521 cggcgtcaaa gccacgcgcg ccgccgacct cacgttcgac gaggcgaagg ctgtcatcga
     5581 catgttcgac gggcccaacg catgagcgcc ccggcgaatt ccgacgcggt ggttgatctg
     5641 caactagcgt acgatcgact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     5701 gggcctttcg ttttatgcca tcatggccgc ggtgatcagc tagccacctg acgtcngggg
     5761 ggggggaaag ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     5821 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     5881 gccatattca acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg
     5941 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     6001 gcttgtatgg gaagccccat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     6061 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     6121 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     6181 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     6241 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     6301 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     6361 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     6421 tgcataatct tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     6481 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     6541 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     6601 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     6661 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     6721 ggcagagcat tacgctgact tgacgggacg gcggctttgt tgaataaatc gaacttttgc
     6781 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca
     6841 aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcaccaacc gtggctccct
     6901 cactttctgg ctggatgatg gggcgattca ggcctggtat gagtcagcaa caccttcttc
     6961 acgaggcaga cctcactagt tccatgagcg tcagaccccg tagaaaagat caaaggatct
     7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7141 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgag
     7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7681 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7741 cgccgcagcc gaacgaccga gcgcaacgcg tgagcccacc agctccgtaa gttcgggcgc
     7801 tgtgtggctc gtacccgcgc attcaggcgg cagggggtct aacgggtcta aggcggcgtg
     7861 tacggccgcc acagcggctc tcagcggccc ggaaacgtcc tcgaaacgac gcatgtgttc
     7921 ctcctggttg gtacaggtgg ttgggggtgc tcggctgtcg ctggtgttcc accaccaggg
     7981 ctcgacggga gagcggggga gtgtgcagtt gtggggtggc ccctcagcga aatatctgac
     8041 ttggagctcg tgtcggacca tacaccggtg attaatcgtg gtctactacc aagcgtgagc
     8101 cacgtcgccg acgaatttga gcagctctgg ctgccgtact ggccgctggc aagcgacgat
     8161 ctgctcgagg ggatctaccg ccaaagccgc gcgtcggccc taggccgccg gtacatcgag
     8221 gcgaacccaa cagcgctggc aaacctgctg gtcgtggacg tagaccatcc agacgcagcg
     8281 ctccgagcgc tcagcgcccg ggggtcccat ccgctgccca acgcgatcgt gggcaatcgc
     8341 gccaacggcc acgcacacgc agtgtgggca ctcaacgccc ctgttccacg caccgaatac
     8401 gcgcggcgta agccgctcgc atacatggcg gcgtgcgccg aaggccttcg gcgcgccgtc
     8461 gatggcgacc gcagttactc aggcctcatg accaaaaacc ccggccacat cgcctgggaa
     8521 acggaatggc tccactcaga tctctacaca ctcagccaca tcgaggccga gctcggcgcg
     8581 aacatgccac cgccgcgctg gcgtcagcag accacgtaca aagcggctcc gacgccgcta
     8641 gggcggaatt gcgcactgtt cgattccgtc aggttgtggg cctatcgtcc cgccctcatg
     8701 cggatctacc tgccgacccg gaacgtggac ggactcggcc gcgcgatcta tgccgagtgc
     8761 cacgcgcgaa acgccgaatt tccgtgcaac gacgtgtgtc c
//
