LOCUS       40924_26356        5166 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJTOOL-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5166)
  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 5166)
  AUTHORS   van Aartsen J, Rajakumar K.
  TITLE     pJTOOL vectors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5166)
  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 5166)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5166)
  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            "Unpublished"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..5166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(58..446)
                     /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(1067..1435)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(1468..1577)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2546..3202)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(3303..4721)
                     /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(join(4722..5166,1))
                     /label=sacB promoter
                     /note="sacB promoter and control region"
ORIGIN
        1 gcggccgcgt tatccgctca caattccaca tgtggaattc cacatgtgga attcccatgt
       61 cagccgttaa gtgttcctgt gtcactcaaa attgctttga gaggctctaa gggcttctca
      121 gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaagct ttaaaagcct
      181 tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa gttgctgatt
      241 tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag agcttagtac
      301 gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa catgagagct
      361 tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac gtactatcaa
      421 caggttgaac tgctgatctt cagatcctct acgccggacg catcgtggcc ggatccagcc
      481 gaccaggctt tccacgcccg cgtgccgctc catgtcgttc gcgcggttct cggaaacgcg
      541 ctgccgcgtt tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg tcgcgcagag
      601 gtcagcgagg gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg tcgggtgaat
      661 cttgttgatg gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg cactgacgcg
      721 ctgatgctcg gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc gcaacgtctc
      781 cgcgtcgggc ttctctcccg cgcggaagct aaccagcagg tgataggtct tgtcggcctc
      841 ggaacgggtg ttgccgtgct gggtcgccat cacctcggcc atgacagcgg gcagggtgtt
      901 tgcctcgcag ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt gctcgtcggt
      961 gatgtacttc accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt
     1021 ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg
     1081 ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca
     1141 gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga
     1201 gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt
     1261 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg
     1321 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc
     1381 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
     1441 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
     1501 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     1561 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     1621 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     1681 tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt ttgtggaata
     1741 tctaccgact ggaaacaggc aaatgcagga aattactgaa ctgaggggac aggcgagaga
     1801 cgatgccaaa gagctacacc gacgagctgg ccgagtgggt tgaatcccgc gcggccaaga
     1861 agcgccggcg tgatgaggct gcggttgcgt tcctggcggt gagggcggat gtcgaggcgg
     1921 cgttagcgtc cggctatgcg ctcgtcacca tttgggagca catgcgggaa acggggaagg
     1981 tcaagttctc ctacgagacg ttccgctcgc acgccaggcg gcacatcaag gccaagcccg
     2041 ccgatgtgcc cgcaccgcag gccaaggctg cggaacccgc gccggcaccc aagacgccgg
     2101 agccacggcg gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct gcggccccga
     2161 ccggcttcac cttcaaccca acaccggaca aaaaggatcc tctacgccgg acgcatcgtg
     2221 gccggcatca ccggcgccac aagtgcggtt gctggcgcct atatcgccga catcaccgat
     2281 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     2341 gcaggccccg tggccggggg actgttgggc gccatctcct tgctgcctcg cgcgtttcgg
     2401 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2461 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2521 gggcgcagcc atgacccggg aattacgccc cgccctgcca ctcatcgcag tactgttgta
     2581 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2641 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2701 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2761 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2821 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2881 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2941 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat
     3001 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     3061 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     3121 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     3181 atccagtgat ttttttctcc attttagctt ccttagctcc tgccctatgg gattcacctt
     3241 tatgttgata agaaataaaa gaaaatgcca ataggatatc ggcattttct tttgcgtttt
     3301 tatttgttaa ctgttaattg tccttgttca aggatgctgt ctttgacaac agatgttttc
     3361 ttgcctttga tgttcagcag gaagcttggc gcaaacgttg attgtttgtc tgcgtagaat
     3421 cctctgtttg tcatatagct tgtaatcacg acattgtttc ctttcgcttg aggtacagcg
     3481 aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat ccatttttaa cacaaggcca
     3541 gttttgttca gcggcttgta tgggccagtt aaagaattag aaacataacc aagcatgtaa
     3601 atatcgttag acgtaatgcc gtcaatcgtc atttttgatc cgcgggagtc agtgaacagg
     3661 taccatttgc cgttcatttt aaagacgttc gcgcgttcaa tttcatctgt tactgtgtta
     3721 gatgcaatca gcggtttcat cacttttttc agtgtgtaat catcgtttag ctcaatcata
     3781 ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat cgctttgcag aagtttttga
     3841 ctttcttgac ggaagaatga tgtgcttttg ccatagtatg ctttgttaaa taaagattct
     3901 tcgccttggt agccatcttc agttccagtg tttgcttcaa atactaagta tttgtggcct
     3961 ttatcttcta cgtagtgagg atctctcagc gtatggttgt cgcctgagct gtagttgcct
     4021 tcatcgatga actgctgtac attttgatac gtttttccgt caccgtcaaa gattgattta
     4081 taatcctcta caccgttgat gttcaaagag ctgtctgatg ctgatacgtt aacttgtgca
     4141 gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat cagtgtagaa taaacggatt
     4201 tttccgtcag atgtaaatgt ggctgaacct gaccattctt gtgtttggtc ttttaggata
     4261 gaatcatttg catcgaattt gtcgctgtct ttaaagacgc ggccagcgtt tttccagctg
     4321 tcaatagaag tttcgccgac tttttgatag aacatgtaaa tcgatgtgtc atccgcattt
     4381 ttaggatctc cggctaatgc aaagacgatg tggtagccgt gatagtttgc gacagtgccg
     4441 tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc cttttgcaga agagatattt
     4501 ttaattgtgg acgaatcgaa ttcaggaact tgatattttt catttttttg ctgttcaggg
     4561 atttgcagca tatcatggcg tgtaatatgg gaaatgccgt atgtttcctt atatggcttt
     4621 tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg ccagcagtgc ggtagtaaag
     4681 gttaatactg ttgcttgttt tgcaaacttt ttgatgttca tcgttcatgt ctcctttttt
     4741 atgtactgtg ttagcggtct gcttcttcca gccctcctgt ttgaagatgg caagttagtt
     4801 acgcacaata aaaaaagacc taaaatatgt aaggggtgac gccaaagtat acactttgcc
     4861 ctttacacat tttaggtctt gcctgcttta tcagtaacaa acccgcgcga tttacttttc
     4921 gacctcattc tattagactc tcgtttggat tgcaactggt ctattttcct cttttgtttg
     4981 atagaaaatc ataaaaggat ttgcagacta cgggcctaaa gaactaaaaa atctatctgt
     5041 ttcttttcat tctctgtatt ttttatagtt tctgttgcat gggcataaag ttgccttttt
     5101 aatcacaatt cagaaaatat cataatatct catttcacta aataatagtg aacggcaggt
     5161 atatgt
//
