LOCUS       Exported                5137 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5243, complete sequence.
ACCESSION   EU024552
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5137)
  AUTHORS   Kvitko BH, Ramos AR, Morello JE, Oh HS, Collmer A.
  TITLE     Identification of harpins in Pseudomonas syringae pv. tomato DC3000,
            which are functionally similar to HrpK1 in promoting translocation 
            of type III secretion system effectors
  JOURNAL   J. Bacteriol. 189 (22), 8059-8072 (2007)
  PUBMED    17873033
REFERENCE   2  (bases 1 to 5137)
  AUTHORS   Kvitko BH, Ramos AR, Morello JE, Oh H-S., Collmer A.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUL-2007) Plant Pathology, Cornell University, 334 
            Plant Science, Ithaca, NY 14853, USA
REFERENCE   3  (bases 1 to 5137)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5137
                     /organism="Cloning vector pCPP5243"
                     /mol_type="other DNA"
                     /db_xref="taxon:458217"
     primer_bind     31..50
                     /label=priming site 1
                     /note="priming site 1"
     misc_recomb     51..98
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(51..98)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(345..1355)
                     /codon_start=1
                     /gene="aadA1"
                     /product="AadA1"
                     /label=aadA1
                     /note="streptomycin/spectinomycin resistance protein"
                     /protein_id="ABS87337.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(345..1355)
                     /gene="aadA1"
                     /label=aadA1
     CDS             complement(345..1136)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             1425..3236
                     /codon_start=1
                     /gene="gus"
                     /product="GUS"
                     /label=gus
                     /note="beta-glucuronidase"
                     /protein_id="ABS87338.1"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            1425..3236
                     /gene="gus"
                     /label=gus
     misc_recomb     3314..3359
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(3314..3347)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(3377..3396)
                     /label=priming site 2
                     /note="priming site 2"
     rep_origin      3393..3779
                     /note="R6K; pir-dependant ori"
     rep_origin      3396..3752
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3849..4709)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ABS87339.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3849..4709)
                     /gene="bla"
                     /label=bla
     promoter        complement(4710..4814)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      4856..5115
                     /regulatory_class="terminator"
                     /note="rrnB"
     terminator      4905..4991
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5083..5110
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"