LOCUS       sequence-13749-e        8749 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 8749)
  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 8749)
  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: pJAK2.D
FEATURES             Location/Qualifiers
     source          1..8749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(110..129)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(194..213)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1260..1848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1749..1768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6852..6869
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6870..6881
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    6894..7018
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(7043..7048,7049..7066,7067..7073)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             7127..7786
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7193..7212)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             8128..8433
                     /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     8312..8331
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(8474..8598)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      8667..8713
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctagccaac aaagcgacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa
       61 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
      121 atgagccata ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat
      181 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
      241 tatcgcttgt atgggaagcc ccatgcgcca gagttgtttc tgaaacatgg caaaggtagc
      301 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
      361 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
      421 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
      481 gttgatgcgc tggcagtgtt cctgcgccgg ttggccaact gtgctggtta tcgccccgtg
      541 ccgcagcgct gtcatcaatc tcgtcgttgt cggttgtggt gtctggcgat gtggtagcag
      601 gattttcgga aggcggcacg gtagtgtcgt cggcttcgtc ggtcgccggc ggagcgatgc
      661 ggaccccgag ttggatctga tgtccgagtc ggcggctgag agcgtcggta atcggggccc
      721 gcagatggcg ctcgatttcg ttttggacaa agctgctcgg cacggataac agagcaaacc
      781 cctcgacgat ggtcaatggc tggacgagat tgagccaagc cctttgctga ggggtcagcg
      841 gagcgctgag attagcatca ctgctgggtc cgtcgtcaac cttagggtcg ccgttaagtt
      901 cggagacgac cgcgttccac actgtggtga agcctgaacc ggggtcatcg gtcaacgacg
      961 tatctccctg gttctcgtta gttccgggtc ttggttccgg gtcgagctac aaccgttgag
     1021 ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag
     1081 ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact
     1141 ttctggctgg atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga
     1201 ggcagacctc actagttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1261 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1321 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1381 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1441 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1501 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1561 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1621 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     1681 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1741 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1801 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1861 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1921 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1981 cgcagccgaa cgaccgagcg caacgcgtga gcccaccagc tccgtaagtt cgggtgctgt
     2041 gtggctcgta cccgcgcatt caggcggcag ggggtctaac gggtctaagg cggcgtgtac
     2101 ggccgccaca gcggctctta gcggcccgga aacgtcctcg aaacgacgca tgtgttcctc
     2161 ctggttggta caggtggttg ggggtgctcg gctgtcgctg gtgtttcatc atcagggctc
     2221 gacgggagag cgggggagtg tgcagttgtg gggtggcccc tcagcgaaat atctgacttg
     2281 gagctcgtgt cggaccatac accggtgatt aatcgtggtt tattatcaag cgtgagccac
     2341 gtcgccgacg aatttgagca gctctggctg ccgtactggt ccctggcaag cgacgatctg
     2401 ctcgagggga tctaccgcca aagccgcgcg tcggccctag gccgccggta catcgaggcg
     2461 aacccaacag cgctggcaaa cctgctggtc gtggacgtag accatccaga cgcagcgctc
     2521 cgagcgctca gcgcccgggg gtcccatccg ctgcccaacg cgatcgtggg caatcgcgcc
     2581 aacggccacg cacacgcagt gtgggcactc aacgcccctg ttccacgcac cgaatacgcg
     2641 cggcgtaagc cgctcgcata catggcggcg tgcgccgaag gccttcggcg cgccgtcgat
     2701 ggcgaccgca gttactcagg cctcatgacc aaaaaccccg gccacatcgc ctgggaaacg
     2761 gaatggctcc actcagatct ctacacactc agccacatcg aggccgagct cggcgcgaac
     2821 atgccaccgc cgcgctggcg tcagcagacc acgtacaaag cggctccgac gccgctaggg
     2881 cggaattgcg cactgttcga ttccgtcagg ttgtgggcct atcttcccgc cctcatgcgg
     2941 atctacctgc cgacccggaa cgtggacgga ctcggccgcg cgatctatgc cgagtgccac
     3001 gcgcgaaacg ccgaatttcc gtgcaacgac gtgtgtcccg gaccgctacc ggacagcgag
     3061 gtccgcgcca tcgccaacag catttggcgt tggatcacaa ccaagtcgcg catttgggcg
     3121 gacgggatcg tggtctacga ggccacactc agtgcgcgcc atgcggccat ctcgcggaag
     3181 ggcgcagcag cgcgcacggc ggcgagcaca gttgcgcggc gcgcaaagtc cgcgtcagcc
     3241 atggaggcat tgctatgagc gacggctaca gcgacggcta cagcgacggc tacaactggc
     3301 agccgactgt ccgcaaaaag cggcgcgtga ccgccgccga aggcgctcga atcaccggac
     3361 tatccgaacg ccacgtcgtc cggctcgtgg cgcaggaacg cagcgagtgg ttcgccgagc
     3421 aggctgcacg ccgcgaacgc atccgcgcct atcacgacga cgagggccac tcttggccgc
     3481 aaacggccaa acatttcggg ctgcatctgg acaccgttaa gcgactcggc tatcgggcga
     3541 ggaaagagcg tgcggcagaa caggaagcgg ctcaaaaggc ccacaacgaa gccgacaatc
     3601 caccgctgtt ctaacgcaat tggggagcgg gtgtcgcggg ggttccgtgg ggggttccgt
     3661 tgcaacgggt cggacaggta aaagtcctgg tagacgctag ttttctggtt tgggccatgc
     3721 ctgtctcgtt gcgtgtttcg ttgcgtccgt tttgaatacc agccagacga gacggggttc
     3781 tacgaatctt ggtcgatacc aagccatttc cgctgaatat cgtggagctc accgccagaa
     3841 tcggtggttg tggtgatgta cgtggcgaac tccgttgtag tgcttgtggt ggcatccgtg
     3901 gcgcggccgc ggtaccagat ctttaaatct agacgagtac ggcgccctgc tgacggatct
     3961 gatccgggaa ccacggcgtg gcaggggatc tcacatcttc gagcgtgcca tcgaacgcgg
     4021 cgagatcgct cccgacgccg atatcgacgt cgcgctcgac gtcctggccg gcgcgcttta
     4081 ctggcggcag tcgatcctcc agacgacggc cgacgacgac tatctcgacc gcctcaccga
     4141 agtcgtactc atggcggtgg ggatggggaa gtgacgcggt ctcaagcgtc gagcgtcgtc
     4201 agcgtgtcga ggatgtcgaa gtcgtagccg tcggcgctgg cgatgtagac ctgctggtcg
     4261 aattgactgt cgcgcataca catcgggccc cggggcccgt cgaacccgac atcgtgcgcg
     4321 gatgccatca ggtccggtat ctcgggggag tgggcccgct ggaagatggc ctcgagcgca
     4381 acgagaccct cgtaacagga ttcggccatc gcgttgagcg gtggcgcgtc ggcgccgtag
     4441 cgggcgacgt agctgcccat caggtccatg gcacccgcgg tggccagtga actgaagtac
     4501 gccgcggcga catagaggtt ttcggtggag ccggcgccgc tggccagcag catgttctcc
     4561 tccatcagcg ggctgaaccg cgccatgcgg tcgtgcccgc cggcgcgcgc gaactcgcgg
     4621 ttgaacaaca cggcgtcctg gccgacgagc agcatcaaca cggcctgcgc ccccgacgcg
     4681 atggccttgc ggaccggtgc gcggaaatcg tcggtgccgt acgggacgta gatctcccgt
     4741 ctgagctcga ggtccagatc tcggcagtac gcgcgggcgg ccgcggcgga acggcgcggc
     4801 cagatgtagt catcgccgac caggcaccag gaccggatgc cgaagtggtc gcgcagccag
     4861 gcgagcgcgg gcgcgatctg gatctgcggt gtctcgcctg tgcagaacac gcccggtgtg
     4921 cgttcaccgc cctcgtacaa cgaggtgtag acgtacggga tgcggtcgcg gaccaccggg
     4981 gagatgcggt tgcgcacggc cgagatgtgc cagccggtca cggcgtcgag accgtgacct
     5041 cgcaaccggt cggcgacggt ccgggcgacg tcgtcgccgg gccgtccgcc gtcgagcacc
     5101 tcgatggtga ccttgcggcc ctgcaggccg cctcggtcgt tgacctcctt ggccgcgagc
     5161 tcggccacgg cctcgcacga aggcgcgaag attcccgctg gcccttgaag cggaatcacc
     5221 agcccgacgc ggaactcaac ctcgccgtcc tgcactccag atcaccgtcg atcccgtgta
     5281 gtctgcgctt caaagctttc tagcagaaat aattcattct gaacagaccc cgccgtcgac
     5341 acgaggagac acccaccatg gccgccggac agcagcgccg ccccaacctc ctgctgccgt
     5401 tggtgcgtct gacccacctc gcggagtcgg cgatcgaacg cgtgctcgcg gactcgtcgc
     5461 tcaagatcga ggactggcgc gtgctcgacg agttggccgg acggcgcacc gtgcccatga
     5521 gcgatctcgc gcaggccacg ctgatcacgg gtccgactct caccagaacc gtcgatcgcc
     5581 ttgtgtcgca agggatcatc taccggactg ccgatctgca tgaccgccgg cgggtgctcg
     5641 tggcgttgac cccgcggggg cggacgctgc gcaaccgcct ggtggacgcg gtagccgagg
     5701 ccgagtgtgc ggcttttgaa tcgtgcgggc tggacgtcga ccagttgcgc gaactcgtcg
     5761 acaccacctc gaatttgact tcgtaaccac ccgcgcccgg ccggcgttca cccttgactt
     5821 ttattttcat ctggatatat ttcgggtgaa tggaaagggg tgaccatgcc gacctacaca
     5881 ttccgttgtt cccactgcgg tcccttcgat ctcacctgcg cgatctccga gcgcgatgcg
     5941 gcggcgacct gtccggagtg ccggacgccg gcgcgccggg tcttcggttc ggtagggctg
     6001 acgacattca ccgcgggaca tcaccgcgca ttcgacgcgg cgtccgcgag cgccgaaagt
     6061 cccacggtgg tgaagtcgat tcccgcaggc gcggaccgcc cgcgggcccc gcgccgcaat
     6121 cccggtctac cgagtctgcc gaggtactag cgacatgggt ggcgtcgggc tcttctacgt
     6181 gggtgcggtg ctcatcatcg acgggctgat gctgctgggc cgcatcagcc cacgaggcgc
     6241 aacaccgctg aacttcttcg tcggcggact gcaggtggtg acgcctacgg tgctgatcct
     6301 gcagtccggc ggagacgcgg ccgtgatctt cgcggcctcc gggctctacc tgttcggctt
     6361 cacctacctg tgggtggcca tcaacaacgt gaccgactgg gacggagaag gtctcggatg
     6421 gttctcgctg ttcgtcgcga tcgccgcact cggctactcg tggcacgcgt tcaccgccga
     6481 ggccgacccg gcgttcgggg tgatctggct gctgtgggca gtgctgtggt tcatgctgtt
     6541 cctgctgctc ggcctggggc acgacgcact ggggcccgcc gtcgggttcg tcgcggtggc
     6601 cgaaggcgtg atcaccgccg ccgtgccggc cttcctgatc gtgtcgggca actgggaaac
     6661 cggcccgctc cccgccgcgg tcatcgccgt gatcggtttt gccgcagttg ttctcgcata
     6721 ccccatcggg cgccgtctcg cagcgccgtc agtcaccaac cctccaccgg ccgcgctcgc
     6781 ggccaccacc cgataagaga aagggagtcc acatgcccga ggtagttttc agcgtcgacc
     6841 attcgggatc gcaccaccac caccaccaca tcgagggccg cggatccaga tcaacaagtt
     6901 tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     6961 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggc
     7021 ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat
     7081 tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     7141 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     7201 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     7261 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     7321 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     7381 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     7441 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     7501 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     7561 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     7621 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     7681 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     7741 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaagat ctggatccgg
     7801 cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat
     7861 atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt
     7921 acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct
     7981 ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga
     8041 aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt
     8101 ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag
     8161 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg
     8221 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     8281 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     8341 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     8401 aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag
     8461 tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt
     8521 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcagc
     8581 tttcttgtac aaagtggttg atctgcagaa ttcgaagctt atcgatgtcg acgtagttaa
     8641 ctagcgtacg atcgactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     8701 cctttcgttt tatctgttgt ttgtccggcc atcatggccg cggtgatca
//
