LOCUS       sequence-13747-e        6389 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 6389)
  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 6389)
  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.B
FEATURES             Location/Qualifiers
     source          1..6389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    232..356
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(381..386,387..404,405..411)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             465..1124
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(531..550)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1466..1771
                     /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     1650..1669
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1812..1936)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2005..2051
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2197..2216)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2281..2300)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3347..3935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3836..3855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcgcaatgg ccaagacaat tgcggatccc ccgagcacac gacgacatac aggacaaagg
       61 ggcacaggta tgacagacgt gagccgaaag attcgagctt ggggacgccg attgatgatc
      121 ggcacggcag cggctgtagt ccttccgggc ctggtggggc ttgccggcgg agcggcaacc
      181 gcgggcgcgt tctcccggcc ggggctgccg gtcgagtacg gatccagatc aacaagtttg
      241 tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
      301 ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg
      361 ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt
      421 tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
      481 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
      541 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
      601 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
      661 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
      721 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
      781 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
      841 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
      901 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
      961 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     1021 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     1081 atgaattaca acagtactgc gatgagtggc agggcggggc gtaaagatct ggatccggct
     1141 tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat
     1201 atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc agcgtattac
     1261 agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt caatatctcc
     1321 ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa
     1381 gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa cggctctttt
     1441 gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata aaagagagag
     1501 ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat
     1561 ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc
     1621 ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc
     1681 ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
     1741 cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc
     1801 tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
     1861 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt
     1921 tcttgtacaa agtggttgat ctgcagaatt cgaagcttat cgatgtcgac gtagttaact
     1981 agcgtacgat cgactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2041 tttcgtttta tctgttgttt gtccggccat catggccgcg gtgatcagct agccaacaaa
     2101 gcgacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2161 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2221 aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg
     2281 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     2341 ggaagcccca tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2401 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2461 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa
     2521 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     2581 cagtgttcct gcgccggttg gccaactgtg ctggttatcg ccccgtgccg cagcgctgtc
     2641 atcaatctcg tcgttgtcgg ttgtggtgtc tggcgatgtg gtagcaggat tttcggaagg
     2701 cggcacggta gtgtcgtcgg cttcgtcggt cgccggcgga gcgatgcgga ccccgagttg
     2761 gatctgatgt ccgagtcggc ggctgagagc gtcggtaatc ggggcccgca gatggcgctc
     2821 gatttcgttt tggacaaagc tgctcggcac ggataacaga gcaaacccct cgacgatggt
     2881 caatggctgg acgagattga gccaagccct ttgctgaggg gtcagcggag cgctgagatt
     2941 agcatcactg ctgggtccgt cgtcaacctt agggtcgccg ttaagttcgg agacgaccgc
     3001 gttccacact gtggtgaagc ctgaaccggg gtcatcggtc aacgacgtat ctccctggtt
     3061 ctcgttagtt ccgggtcttg gttccgggtc gagctacaac cgttgagttg aaggatcaga
     3121 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     3181 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     3241 atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcact
     3301 agttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3361 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3421 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3481 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3541 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3601 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3661 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3721 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3781 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3841 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3901 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3961 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4021 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4081 ccgagcgcaa cgcgtgagcc caccagctcc gtaagttcgg gtgctgtgtg gctcgtaccc
     4141 gcgcattcag gcggcagggg gtctaacggg tctaaggcgg cgtgtacggc cgccacagcg
     4201 gctcttagcg gcccggaaac gtcctcgaaa cgacgcatgt gttcctcctg gttggtacag
     4261 gtggttgggg gtgctcggct gtcgctggtg tttcatcatc agggctcgac gggagagcgg
     4321 gggagtgtgc agttgtgggg tggcccctca gcgaaatatc tgacttggag ctcgtgtcgg
     4381 accatacacc ggtgattaat cgtggtttat tatcaagcgt gagccacgtc gccgacgaat
     4441 ttgagcagct ctggctgccg tactggtccc tggcaagcga cgatctgctc gaggggatct
     4501 accgccaaag ccgcgcgtcg gccctaggcc gccggtacat cgaggcgaac ccaacagcgc
     4561 tggcaaacct gctggtcgtg gacgtagacc atccagacgc agcgctccga gcgctcagcg
     4621 cccgggggtc ccatccgctg cccaacgcga tcgtgggcaa tcgcgccaac ggccacgcac
     4681 acgcagtgtg ggcactcaac gcccctgttc cacgcaccga atacgcgcgg cgtaagccgc
     4741 tcgcatacat ggcggcgtgc gccgaaggcc ttcggcgcgc cgtcgatggc gaccgcagtt
     4801 actcaggcct catgaccaaa aaccccggcc acatcgcctg ggaaacggaa tggctccact
     4861 cagatctcta cacactcagc cacatcgagg ccgagctcgg cgcgaacatg ccaccgccgc
     4921 gctggcgtca gcagaccacg tacaaagcgg ctccgacgcc gctagggcgg aattgcgcac
     4981 tgttcgattc cgtcaggttg tgggcctatc ttcccgccct catgcggatc tacctgccga
     5041 cccggaacgt ggacggactc ggccgcgcga tctatgccga gtgccacgcg cgaaacgccg
     5101 aatttccgtg caacgacgtg tgtcccggac cgctaccgga cagcgaggtc cgcgccatcg
     5161 ccaacagcat ttggcgttgg atcacaacca agtcgcgcat ttgggcggac gggatcgtgg
     5221 tctacgaggc cacactcagt gcgcgccatg cggccatctc gcggaagggc gcagcagcgc
     5281 gcacggcggc gagcacagtt gcgcggcgcg caaagtccgc gtcagccatg gaggcattgc
     5341 tatgagcgac ggctacagcg acggctacag cgacggctac aactggcagc cgactgtccg
     5401 caaaaagcgg cgcgtgaccg ccgccgaagg cgctcgaatc accggactat ccgaacgcca
     5461 cgtcgtccgg ctcgtggcgc aggaacgcag cgagtggttc gccgagcagg ctgcacgccg
     5521 cgaacgcatc cgcgcctatc acgacgacga gggccactct tggccgcaaa cggccaaaca
     5581 tttcgggctg catctggaca ccgttaagcg actcggctat cgggcgagga aagagcgtgc
     5641 ggcagaacag gaagcggctc aaaaggccca caacgaagcc gacaatccac cgctgttcta
     5701 acgcaattgg ggagcgggtg tcgcgggggt tccgtggggg gttccgttgc aacgggtcgg
     5761 acaggtaaaa gtcctggtag acgctagttt tctggtttgg gccatgcctg tctcgttgcg
     5821 tgtttcgttg cgtccgtttt gaataccagc cagacgagac ggggttctac gaatcttggt
     5881 cgataccaag ccatttccgc tgaatatcgt ggagctcacc gccagaatcg gtggttgtgg
     5941 tgatgtacgt ggcgaactcc gttgtagtgc ttgtggtggc atccgtggcg cggccgcggt
     6001 accagatctt taaatctaga ggtgaccaca acgacgcgcc cgctttgatc ggggacgtct
     6061 gcggccgacc atttacgggt cttgttgtcg ttggcggtca tgggccgaac atactcaccc
     6121 ggatcggagg gccgaggaca aggtcgaacg aggggcatga cccggtgcgg ggcttcttgc
     6181 actcggcata ggcgagtgct aagaataacg ttggcactcg cgaccggtga gtcgtaggtc
     6241 gggacggtga ggccaggccc gtcgtcgcag cgagtggcag cgaggacaac ttgagccgtc
     6301 cgtcgcgggc actgcgcccg gccagcgtaa gtagcggggt tgccgtcacc cggtgacccc
     6361 cggtttcatc cccgatccgg aggaatcac
//
