LOCUS       40924_26924        6671 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pKNG202, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Spagnolo J, Bigot S, Denis Y, Bordi C, de Bentzmann S.
  TITLE     Development of a genetic tool for activating chromosomal expression 
            of cryptic or tightly regulated loci in Pseudomonas aeruginosa
  JOURNAL   Plasmid (2011) In press
  PUBMED    22212534
REFERENCE   2  (bases 1 to 6671)
  AUTHORS   Spagnolo J, Bigot S, Bordi C, de Bentzmann S.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2011) Institut de Microbiologie de la 
            Mediterrannee, UMR 7255 Aix-Marseille University, 31 chemin Joseph 
            Aiguier, Marseille, Bouches du Rhone 13402, France
REFERENCE   3  (bases 1 to 6671)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6671)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 
            (2011) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-DEC-2011) Institut de Microbiologie de la Mediterrannee, UMR 
            7255 Aix-Marseille University, 31 chemin Joseph Aiguier, Marseille, 
            Bouches du Rhone 13402, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        141..169
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             693..1184
                     /codon_start=1
                     /gene="strA"
                     /product="StrA"
                     /function="selection marker for integration"
                     /label=strA
                     /protein_id="AFF18865.1"
                     /translation="MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPD
                     EDKSTPLHDLLARVERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRL
                     GTADRYADLALMIANAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG
                     "
     gene            693..1184
                     /gene="strA"
                     /label=strA
     CDS             1184..2020
                     /codon_start=1
                     /gene="strB"
                     /product="StrB"
                     /function="selection marker for integration"
                     /label=strB
                     /protein_id="AFF18866.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            1184..2020
                     /gene="strB"
                     /label=strB
     rep_origin      2056..2444
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            3164..3273
                     /label=oriT
                     /note="incP origin of transfer"
     mobile_element  3886..4186
                     /mobile_element_type="insertion sequence:IS1"
                     /label=insertion sequence:IS1
                     /note="from Escherichia coli"
     CDS             complement(4330..5748)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(5749..6194)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    6255..6654
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
ORIGIN
        1 gtcgacggat ccgctagcgg ccggccacta gtctcgagat gcatagatct gggccctcta
       61 gtatagtcta tagtccgtgg aattattata tttatctccg acgatattct catcagtgaa
      121 atccagggga attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta
      181 tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc
      241 gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg
      301 ccgggcctct tgcgggataa ctgcacattc gggatatttc tctatattcg cgcttcatca
      361 gaaaactgaa ggaacctcca ttgaatcgaa ctaatatttt ttttggtgaa tcgcattctg
      421 actggttgcc tgtcagaggc ggagaatctg gtgattttgt ttttcgacgt ggtgacgggc
      481 atgccttcgc gaaaatcgca cctgcttccc gccgcggtga gctcgctgga gagcgtgacc
      541 gcctcatttg gctcaaaggt cgaggtgtgg cttgccccga ggtcatcaac tggcaggagg
      601 aacaggaggg tgcatgcttg gtgataacgg caattccggg agtaccggcg gctgatctgt
      661 ctggagcgga tttgctcaaa gcgtggccgt caatggggca gcaacttggc gctgttcaca
      721 gcctatcggt tgatcaatgt ccgtttgagc gcaggctgtc gcgaatgttc ggacgcgccg
      781 ttgatgtggt gtcccgcaat gccgtcaatc ccgacttctt accggacgag gacaagagta
      841 cgccgctgca cgatcttttg gctcgtgtcg aacgagagct accggtgcgg ctcgaccaag
      901 agcgcaccga tatggttgtt tgccatggtg atccctgcat gccgaacttc atggtggacc
      961 ctaaaactct tcaatgcacg ggtctgatcg accttgggcg gctcggaaca gcagatcgct
     1021 atgccgattt ggcactcatg attgctaacg ccgaagagaa ctgggcagcg ccagatgaag
     1081 cagagcgcgc cttcgctgtc ctattcaatg tattggggat cgaagccccc gaccgcgaac
     1141 gccttgcctt ctatctgcga ttggaccctc tgacttgggg ttgatgttca tgccgcctgt
     1201 ttttcctgct cattggcacg tttcgcaacc tgttctcatt gcggacacct tttccagcct
     1261 cgtttggaaa gtttcattgc cagacgggac tcctgcaatc gtcaagggat tgaaacctat
     1321 agaagacatt gctgatgaac tgcgcggggc cgactatctg gtatggcgca atgggagggg
     1381 agcagtccgg ttgctcggtc gtgagaacaa tctgatgttg ctcgaatatg ccggggagcg
     1441 aatgctctct cacatcgttg ccgagcacgg cgactaccag gcgaccgaaa ttgcagcgga
     1501 actaatggcg aagctgtatg ccgcatctga ggaacccctg ccttctgccc ttctcccgat
     1561 ccgggatcgc tttgcagctt tgtttcagcg ggcgcgcgat gatcaaaacg caggttgtca
     1621 aactgactac gtccacgcgg cgattatagc cgatcaaatg atgagcaatg cctcggaact
     1681 gcgtgggcta catggcgatc tgcatcatga aaacatcatg ttctccagtc gcggctggct
     1741 ggtgatagat cccgtcggtc tggtcggtga agtgggcttt ggcgccgcca atatgttcta
     1801 cgatccggct gacagagacg acctttgtct cgatcctaga cgcattgcac agatggcgga
     1861 cgcattctct cgtgcgctgg acgtcgatcc gcgtcgcctg ctcgaccagg cgtacgctta
     1921 tgggtgcctt tccgcagctt ggaacgcgga tggagaagag gagcaacgcg atctagctat
     1981 cgcggccgcg atcaagcagg tgcgacagac gtcatactag atatctagat ccggcccgat
     2041 gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta cgtactaagc
     2101 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
     2161 ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc tctcatgttt
     2221 cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa cttaaatagc
     2281 ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta aagcttttaa
     2341 ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc cttagagcct
     2401 ctcaaagcaa ttttcagtga cacaggaaca cttaacggct gacatgggaa ttctactgtt
     2461 tgggtgtatg agctatttgc aaaggaaatt ggtgatgaca aagctcggcg ctatttgaag
     2521 aaaatcgact atggcaacgc cgatcctttt tgtccggtgt tgggttgaag gtgaagccgg
     2581 tcggggccgc agcgggggcc ggcttttcag ccttgccccc ctgcttcggc cgccgtggct
     2641 ccggcgtctt gggtgccggc gcgggttccg cagccttggc ctgcggtgcg ggcacatcgg
     2701 cgggcttggc cttgatgtgc cgcctggcgt gcgagcggaa cgtctcgtag gagaacttga
     2761 ccttccccgt ttcccgcatg tgctcccaaa tggtgacgag cgcatagccg gacgctaacg
     2821 ccgcctcgac atccgccctc accgccagga acgcaaccgc agcctcatca cgccggcgct
     2881 tcttggccgc gcgggattca acccactcgg ccagctcgtc ggtgtagctc tttggcatcg
     2941 tctctcgcct gtcccctcag ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga
     3001 tattccacaa aacagcaggg aagcagcgct tttccgctgc ataaccctgc ttcggggtca
     3061 ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa
     3121 gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag
     3181 taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc
     3241 tcaacgggaa tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca
     3301 agcggatggc tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact
     3361 gccttccaga cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt
     3421 cggcctacct gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc
     3481 acgtccgcga gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac
     3541 tctggctcac cgacgacccg cgcacggcgc ggttcggtga tgccacgatc cgggcaacgt
     3601 tgttgccatt gctgcagttc acttacaccg cttctcaacc cggtacgcac cataaaatca
     3661 ttgatatggc catgaatggc gttggatgcc gggcaacagc ccgcattatg ggcgttggcc
     3721 tcaacacgat tttacgtcac ttaaaaaact caggccgcag tcggtaacct cgcgcataca
     3781 gccgggcagt gacgtcatcg tctgcgcgga aatggacgaa cagtggggct atgtcggggc
     3841 taaatcgcgc cagcgctggc tgttttacgc gtatgacagt ctccggaaga cggttgttgc
     3901 gcacgtattc ggtgaacgca ctatggcgac gctggggcgt cttatgagcc tgctgtcacc
     3961 ctttgacgtg gtgatatgga tgacggatgg ctggccgctg tatgaatccc gcctgaaggg
     4021 aaagctgcac gtaatcagca agcgatatac gcagcgaatt gagcggcata acctgaatct
     4081 gaggcagcac ctggcacggc tgggacggaa gtcgctgtcg ttctcaaaat cggtggagct
     4141 gcatgacaaa gtcatcgggc attatctgaa cataaaacac tatcaataag ttggagtcat
     4201 taccaaaagg ttaggaatac ggttagccat ttgcctgctt ttatatagtt catatgggat
     4261 tcacctttat gttgataaga aataaaagaa aatgccaata ggatatcggc attttctttt
     4321 gcgtttttat ttgttaactg ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga
     4381 tgttttcttg cctttgatgt tcagcaggaa gcttggcgca aacgttgatt gtttgtctgc
     4441 gtagaatcct ctgtttgtca tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg
     4501 tacagcgaag tgtgagtaag taaaggttac atcgttagga tcaagatcca tttttaacac
     4561 aaggccagtt ttgttcagcg gcttgtatgg gccagttaaa gaattagaaa cataaccaag
     4621 catgtaaata tcgttagacg taatgccgtc aatcgtcatt tttgatccgc gggagtcagt
     4681 gaacaggtac catttgccgt tcattttaaa gacgttcgcg cgttcaattt catctgttac
     4741 tgtgttagat gcaatcagcg gtttcatcac ttttttcagt gtgtaatcat cgtttagctc
     4801 aatcataccg agagcgccgt ttgctaactc agccgtgcgt tttttatcgc tttgcagaag
     4861 tttttgactt tcttgacgga agaatgatgt gcttttgcca tagtatgctt tgttaaataa
     4921 agattcttcg ccttggtagc catcttcagt tccagtgttt gcttcaaata ctaagtattt
     4981 gtggccttta tcttctacgt agtgaggatc tctcagcgta tggttgtcgc ctgagctgta
     5041 gttgccttca tcgatgaact gctgtacatt ttgatacgtt tttccgtcac cgtcaaagat
     5101 tgatttataa tcctctacac cgttgatgtt caaagagctg tctgatgctg atacgttaac
     5161 ttgtgcagtt gtcagtgttt gtttgccgta atgtttaccg gagaaatcag tgtagaataa
     5221 acggattttt ccgtcagatg taaatgtggc tgaacctgac cattcttgtg tttggtcttt
     5281 taggatagaa tcatttgcat cgaatttgtc gctgtcttta aagacgcggc cagcgttttt
     5341 ccagctgtca atagaagttt cgccgacttt ttgatagaac atgtaaatcg atgtgtcatc
     5401 cgcattttta ggatctccgg ctaatgcaaa gacgatgtgg tagccgtgat agtttgcgac
     5461 agtgccgtca gcgttttgta atggccagct gtcccaaacg tccaggcctt ttgcagaaga
     5521 gatattttta attgtggacg aatcgaattc aggaacttga tatttttcat ttttttgctg
     5581 ttcagggatt tgcagcatat catggcgtgt aatatgggaa atgccgtatg tttccttata
     5641 tggcttttgg ttcgtttctt tcgcaaacgc ttgagttgcg cctcctgcca gcagtgcggt
     5701 agtaaaggtt aatactgttg cttgttttgc aaactttttg atgttcatcg ttcatgtctc
     5761 cttttttatg tactgtgtta gcggtctgct tcttccagcc ctcctgtttg aagatggcaa
     5821 gttagttacg cacaataaaa aaagacctaa aatatgtaag gggtgacgcc aaagtataca
     5881 ctttgccctt tacacatttt aggtcttgcc tgctttatca gtaacaaacc cgcgcgattt
     5941 acttttcgac ctcattctat tagactctcg tttggattgc aactggtcta ttttcctctt
     6001 ttgtttgata gaaaatcata aaaggatttg cagactacgg gcctaaagaa ctaaaaaatc
     6061 tatctgtttc ttttcattct ctgtattttt tatagtttct gttgcatggg cataaagttg
     6121 cctttttaat cacaattcag aaaatatcat aatatctcat ttcactaaat aatagtgaac
     6181 ggcaggtata tgtgatgggt taaaaaggat cttaaggcct aggtctagct tggctgcagg
     6241 gggggggggg ggacttccat tgttcattcc acggacaaaa acagagaaag gaaacgacag
     6301 aggccaaaaa gcctcgcttt cagcacctgt cgtttccttt cttttcagag ggtattttaa
     6361 ataaaaacat taagttatga cgaagaagaa cggaaacgcc ttaaaccgga aaattttcat
     6421 aaatagcgaa aacccgcgag gtcgccgccc cgtaacctgt cggatcaccg gaaaggaccc
     6481 gtaaagtgat aatgattatc atctacatat cacaacgtgc gtggaggcca tcaaaccacg
     6541 tcaaataatc aattatgacg caggtatcgt attaattgat ctgcatcaac ttaacgtaaa
     6601 aacaacttca gacaatacaa atcagcgaca ctgaatacgg ggcaacctca tgtccccccc
     6661 cccccctgca g
//
