LOCUS       Exported                6671 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pKNG202, complete sequence.
ACCESSION   JQ282797
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6671
                     /organism="Shuttle vector pKNG202"
                     /mol_type="other DNA"
                     /label=derived from vector pKNG101
                     /PCR_primers="fwd_name: upkn, fwd_seq:
                     ccctggatttcactgatgag, rev_name: rpkn, rev_seq: 
                     catatcacaacgtgcgtgga"
                     /note="derived from vector pKNG101"
                     /db_xref="taxon:1165853"
     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      2050..2444
                     /note="origin of replication of plasmid R6K (defective pir 
                     gene)"
     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"
     rep_origin      3162..3272
                     /label=RK6 conjugational DNA transfer origin
                     /note="RK6 conjugational DNA transfer origin"
     oriT            3164..3273
                     /note="incP origin of transfer"
     mobile_element  3886..4186
                     /mobile_element_type="insertion sequence:IS1"
                     /note="from Escherichia coli"
     CDS             complement(4327..5748)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="from Bacillus subtilis levansucrase"
                     /protein_id="AFF18867.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(4327..5748)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(5749..6194)
                     /gene="sacR"
                     /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"