LOCUS       40924_26361        5344 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJTOOL-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5344)
  AUTHORS   van Aartsen JJ, Rajakumar K.
  TITLE     An optimized method for suicide vector-based allelic exchange in 
            Klebsiella pneumoniae
  JOURNAL   J. Microbiol. Methods 86 (3), 313-319 (2011)
  PUBMED    21699924
REFERENCE   2  (bases 1 to 5344)
  AUTHORS   van Aartsen J, Rajakumar K.
  TITLE     pJTOOL vectors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5344)
  AUTHORS   van Aartsen J, Rajakumar K.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2011) Department of Infection, Immunity and 
            Inflammation, University of Leicester, United Kingdom, University 
            Road, Leicester, Leicestershire LE1 9HN, United Kingdom
REFERENCE   4  (bases 1 to 5344)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5344)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2011"; volume: "86"; issue: "3"; pages: 
            "313-319"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (01-APR-2011) Department of Infection, Immunity and Inflammation, 
            University of Leicester, United Kingdom, University Road, Leicester,
            Leicestershire LE1 9HN, United Kingdom"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..418)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1039..1407)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(1440..1549)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2518..3174)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(3275..4693)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(4694..5139)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     promoter        5159..5177
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5186..5293
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(5306..5324)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(5323..5339)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 catgtggaat tccacatgtg gaattcccat gtcagccgtt aagtgttcct gtgtcactca
       61 aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca
      121 caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact
      181 taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag
      241 agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc
      301 catgagggtt tagttcgtta aacatgagag cttagtacgt taaacatgag agcttagtac
      361 gtgaaacatg agagcttagt acgtactatc aacaggttga actgctgatc ttcagatcct
      421 ctacgccgga cgcatcgtgg ccggatccag ccgaccaggc tttccacgcc cgcgtgccgc
      481 tccatgtcgt tcgcgcggtt ctcggaaacg cgctgccgcg tttcgtgatt gtcacgctca
      541 agcccgtagt cccgttcgag cgtcgcgcag aggtcagcga gggcgcggta ggcccgatac
      601 ggctcatgga tggtgtttcg ggtcgggtga atcttgttga tggcgatatg gatgtgcagg
      661 ttgtcggtgt cgtgatgcac ggcactgacg cgctgatgct cggcgaagcc aagcccagcg
      721 cagatgcggt cctcaatcgc gcgcaacgtc tccgcgtcgg gcttctctcc cgcgcggaag
      781 ctaaccagca ggtgataggt cttgtcggcc tcggaacggg tgttgccgtg ctgggtcgcc
      841 atcacctcgg ccatgacagc gggcagggtg tttgcctcgc agttcgtgac gcgcacgtga
      901 cccaggcgct cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg
      961 ctcttcttga tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt
     1021 tagcggctaa aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc
     1081 aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa
     1141 ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag
     1201 gtcgccattg atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt
     1261 gtagccctgg ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt
     1321 ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt
     1381 cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc
     1441 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt
     1501 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     1561 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     1621 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga
     1681 aaagcgctgc ttccctgctg ttttgtggaa tatctaccga ctggaaacag gcaaatgcag
     1741 gaaattactg aactgagggg acaggcgaga gacgatgcca aagagctaca ccgacgagct
     1801 ggccgagtgg gttgaatccc gcgcggccaa gaagcgccgg cgtgatgagg ctgcggttgc
     1861 gttcctggcg gtgagggcgg atgtcgaggc ggcgttagcg tccggctatg cgctcgtcac
     1921 catttgggag cacatgcggg aaacggggaa ggtcaagttc tcctacgaga cgttccgctc
     1981 gcacgccagg cggcacatca aggccaagcc cgccgatgtg cccgcaccgc aggccaaggc
     2041 tgcggaaccc gcgccggcac ccaagacgcc ggagccacgg cggccgaagc aggggggcaa
     2101 ggctgaaaag ccggcccccg ctgcggcccc gaccggcttc accttcaacc caacaccgga
     2161 caaaaaggat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaagtgcgg
     2221 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     2281 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     2341 gcgccatctc cttgctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2401 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2461 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgacccg ggaattacgc
     2521 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     2581 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     2641 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     2701 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     2761 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     2821 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     2881 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     2941 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     3001 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     3061 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     3121 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     3181 ttccttagct cctgccctat gggattcacc tttatgttga taagaaataa aagaaaatgc
     3241 caataggata tcggcatttt cttttgcgtt tttatttgtt aactgttaat tgtccttgtt
     3301 caaggatgct gtctttgaca acagatgttt tcttgccttt gatgttcagc aggaagcttg
     3361 gcgcaaacgt tgattgtttg tctgcgtaga atcctctgtt tgtcatatag cttgtaatca
     3421 cgacattgtt tcctttcgct tgaggtacag cgaagtgtga gtaagtaaag gttacatcgt
     3481 taggatcaag atccattttt aacacaaggc cagttttgtt cagcggcttg tatgggccag
     3541 ttaaagaatt agaaacataa ccaagcatgt aaatatcgtt agacgtaatg ccgtcaatcg
     3601 tcatttttga tccgcgggag tcagtgaaca ggtaccattt gccgttcatt ttaaagacgt
     3661 tcgcgcgttc aatttcatct gttactgtgt tagatgcaat cagcggtttc atcacttttt
     3721 tcagtgtgta atcatcgttt agctcaatca taccgagagc gccgtttgct aactcagccg
     3781 tgcgtttttt atcgctttgc agaagttttt gactttcttg acggaagaat gatgtgcttt
     3841 tgccatagta tgctttgtta aataaagatt cttcgccttg gtagccatct tcagttccag
     3901 tgtttgcttc aaatactaag tatttgtggc ctttatcttc tacgtagtga ggatctctca
     3961 gcgtatggtt gtcgcctgag ctgtagttgc cttcatcgat gaactgctgt acattttgat
     4021 acgtttttcc gtcaccgtca aagattgatt tataatcctc tacaccgttg atgttcaaag
     4081 agctgtctga tgctgatacg ttaacttgtg cagttgtcag tgtttgtttg ccgtaatgtt
     4141 taccggagaa atcagtgtag aataaacgga tttttccgtc agatgtaaat gtggctgaac
     4201 ctgaccattc ttgtgtttgg tcttttagga tagaatcatt tgcatcgaat ttgtcgctgt
     4261 ctttaaagac gcggccagcg tttttccagc tgtcaataga agtttcgccg actttttgat
     4321 agaacatgta aatcgatgtg tcatccgcat ttttaggatc tccggctaat gcaaagacga
     4381 tgtggtagcc gtgatagttt gcgacagtgc cgtcagcgtt ttgtaatggc cagctgtccc
     4441 aaacgtccag gccttttgca gaagagatat ttttaattgt ggacgaatcg aattcaggaa
     4501 cttgatattt ttcatttttt tgctgttcag ggatttgcag catatcatgg cgtgtaatat
     4561 gggaaatgcc gtatgtttcc ttatatggct tttggttcgt ttctttcgca aacgcttgag
     4621 ttgcgcctcc tgccagcagt gcggtagtaa aggttaatac tgttgcttgt tttgcaaact
     4681 ttttgatgtt catcgttcat gtctcctttt ttatgtactg tgttagcggt ctgcttcttc
     4741 cagccctcct gtttgaagat ggcaagttag ttacgcacaa taaaaaaaga cctaaaatat
     4801 gtaaggggtg acgccaaagt atacactttg ccctttacac attttaggtc ttgcctgctt
     4861 tatcagtaac aaacccgcgc gatttacttt tcgacctcat tctattagac tctcgtttgg
     4921 attgcaactg gtctattttc ctcttttgtt tgatagaaaa tcataaaagg atttgcagac
     4981 tacgggccta aagaactaaa aaatctatct gtttcttttc attctctgta ttttttatag
     5041 tttctgttgc atgggcataa agttgccttt ttaatcacaa ttcagaaaat atcataatat
     5101 ctcatttcac taaataatag tgaacggcag gtatatgtga ttaccctgtt atccctagta
     5161 atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgct ctagaactag
     5221 tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgaggg
     5281 ggggcccggt acccagcttt tgttcccttt agtgagggtt aattgttatc cgctcacaat
     5341 tcca
//
