LOCUS       sequence-13746-e        6354 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 6354)
  AUTHORS   Sun J, Lau A, Wang X, Liao TY, Zoubeidi A, Hmama Z
  TITLE     A broad-range of recombination cloning vectors in mycobacteria.
  JOURNAL   Plasmid. 2009 Nov . 62(3):158-65.
  PUBMED    19647016
REFERENCE   2  (bases 1 to 6354)
  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: "Plasmid. 
            2009 Nov . 62(3):158-65."
COMMENT     Sequence Label: pJAK1.B
FEATURES             Location/Qualifiers
     source          1..6354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..84)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             76..891
                     /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(149..168)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1223..1811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1712..1731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4454..4578
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4603..4608,4609..4626,4627..4633)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4687..5346
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4753..4772)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5688..5993
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5872..5891
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(6034..6158)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      6227..6273
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
       61 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta
      121 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
      181 tcaggtgcga caatctatcg cttgtatggg aagccccatg cgccagagtt gtttctgaaa
      241 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
      301 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
      361 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
      421 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
      481 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
      541 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
      601 gaacaagtct ggaaagaaat gcataatctt ttgccattct caccggattc agtcgtcact
      661 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
      721 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
      781 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
      841 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
      901 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt
      961 gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag
     1021 accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc
     1081 tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg
     1141 agtcagcaac accttcttca cgaggcagac ctcactagtt ccactgagcg tcagaccccg
     1201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     1621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1861 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1921 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcaacgcg tgcggccgcg
     1981 gtacccgggg atcctctaga gtcgaccacc aagggcacca tctctgcttg ggccaccccg
     2041 ttggccgcag ccagctcgct gagagccgtg aacgacaggg cgaacgccag cccgccgacg
     2101 gcgagggttc cgaccgctgc aactcccggt gcaaccttgt cccggtctat tctcttcact
     2161 gcaccagctc caatctggtg tgaatgcccc tcgtctgttc gcgcaggcgg ggggctctat
     2221 tcgtttgtca gcatcgaaag tagccagatc agggatgcgt tgcaaccgcg tatgcccagg
     2281 tcagaagagt cgcacaagag ttgcagaccc ctggaaagaa aaatggccag agggcgaaaa
     2341 caccctctga ccagcggagc gggcgacggg aatcgaaccc gcgtagctag tttggaagaa
     2401 tgggtgtctg ccgaccacat atgggccggt caagataggt ttttaccccc tctcggctgc
     2461 atcctctaag tggaaagaaa ttgcaggtcg tagaagcgcg ttgaagcctg agagttgcac
     2521 aggagttgca acccggtagc cttgttcacg acgagaggag acctagttgg cacgtcgcgg
     2581 atggggatcg ctgaagactc agcgcagcgg gaggatccaa gcctcatacg tcaacccgca
     2641 ggacggtgtg aggtactacg cgctgcagac ctacgacaac aagatggacg ccgaagcctg
     2701 gctcgcgggc gagaagcggc tcatcgagat ggagacctgg acccctccac aggaccgggc
     2761 gaagaaggca gccgccagcg ccatcacgct ggaggagtac acccggaagt ggctcgtgga
     2821 gcgcgacctc gcagacggca ccagggatct gtacagcggg cacgcggagc gccgcatcta
     2881 cccggtgcta ggtgaagtgg cggtcacaga gatgacgcca gctctggtgc gtgcgtggtg
     2941 ggccgggatg ggtaggaagc acccgactgc ccgccggcat gcctacaacg tcctccgggc
     3001 ggtgatgaac acagcggtcg aggacaagct gatcgcagag aacccgtgcc ggatcgagca
     3061 gaaggcagcc gatgagcgcg acgtagaggc gctgacgcct gaggagctgg acatcgtcgc
     3121 cgctgagatc ttcgagcact accggatcgc ggcatacatc ctggcgtgga cgagcctccg
     3181 gttcggagag ctgatcgagc ttcgccgcaa ggacatcgtg gacgacggca tgacgatgaa
     3241 gctccgggtg cgccgtggcg cttcccgcgt ggggaacaag atcgtcgttg gcaacgccaa
     3301 gaccgtccgg tcgaagcgtc ctgtgacggt tccgcctcac gtcgcggaga tgatccgagc
     3361 gcacatgaag gaccgtacga agatgaacaa gggccccgag gcattcctgg tgaccacgac
     3421 gcagggcaac cggctgtcga agtccgcgtt caccaagtcg ctgaagcgtg gctacgccaa
     3481 gatcggtcgg ccggaactcc gcatccacga cctccgcgct gtcggcgcta cgttcgccgc
     3541 tcaggcaggt gcgacgacca aggagctgat ggcccgtctc ggtcacacga ctcctaggat
     3601 ggcgatgaag taccagatgg cgtctgaggc ccgcgacgag gctatcgctg aggcgatgtc
     3661 caagctggcc aagacctcct gaaacgcaaa aagcccccct cccaaggaca ctgagtccta
     3721 aagagggggg tttcttgtca gtacgcgaag aaccacgcct ggccgcgagc gccagcaccg
     3781 ccgctctgtg cggagacctg ggcaccagcc ccgccgccgc caggagcatt gccgttcccg
     3841 ccagaaatct agaggtgacc acaacgacgc gcccgctttg atcggggacg tctgcggccg
     3901 accatttacg ggtcttgttg tcgttggcgg tcatgggccg aacatactca cccggatcgg
     3961 agggccgagg acaaggtcga acgaggggca tgacccggtg cggggcttct tgcactcggc
     4021 ataggcgagt gctaagaata acgttggcac tcgcgaccgg tgagtcgtag gtcgggacgg
     4081 tgaggccagg cccgtcgtcg cagcgagtgg cagcgaggac aacttgagcc gtccgtcgcg
     4141 ggcactgcgc ccggccagcg taagtagcgg ggttgccgtc acccggtgac ccccggtttc
     4201 atccccgatc cggaggaatc acttcgcaat ggccaagaca attgcggatc ccccgagcac
     4261 acgacgacat acaggacaaa ggggcacagg tatgacagac gtgagccgaa agattcgagc
     4321 ttggggacgc cgattgatga tcggcacggc agcggctgta gtccttccgg gcctggtggg
     4381 gcttgccggc ggagcggcaa ccgcgggcgc gttctcccgg ccggggctgc cggtcgagta
     4441 cggatccaga tcaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
     4501 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
     4561 acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc
     4621 ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa
     4681 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     4741 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     4801 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     4861 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     4921 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     4981 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     5041 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     5101 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     5161 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     5221 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     5281 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     5341 gcgtaaagat ctggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
     5401 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
     5461 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     5521 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
     5581 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     5641 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
     5701 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     5761 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     5821 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     5881 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
     5941 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
     6001 gctcccttat acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta
     6061 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     6121 ttttacgttt ctcgttcagc tttcttgtac aaagtggttg atctgcagaa ttcgaagctt
     6181 atcgatgtcg acgtagttaa ctagcgtacg atcgactgcc aggcatcaaa taaaacgaaa
     6241 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtccggcc atcatggccg
     6301 cggtgatcag ctagccaaca aagcgacgtt gtgtctcaaa atctctgatg ttac
//
