LOCUS       Exported               12904 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5264, complete sequence.
ACCESSION   EU024542
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12904)
  AUTHORS   Wei CF, Kvitko BH, Shimizu R, Crabill E, Alfano JR, Lin NC, Martin 
            GB, Huang HC, Collmer A.
  TITLE     A Pseudomonas syringae pv. tomato DC3000 mutant lacking the type III
            effector HopQ1-1 is able to cause disease in the model plant 
            Nicotiana benthamiana
  JOURNAL   Plant J. 51 (1), 32-46 (2007)
  PUBMED    17559511
REFERENCE   2  (bases 1 to 12904)
  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   3  (bases 1 to 12904)
  AUTHORS   Kvitko BH, Ramos AR, Morello JE, Oh H-S., Collmer A.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2007) Plant Pathology, Cornell University, 334 
            Plant Science, Ithaca, NY 14853, USA
REFERENCE   4  (bases 1 to 12904)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12904
                     /organism="Cloning vector pCPP5264"
                     /mol_type="other DNA"
                     /db_xref="taxon:458220"
     misc_recomb     complement(13..12897)
                     /label=attB2
                     /note="attB2"
     primer_bind     complement(65..81)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    89..105
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(113..143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    158..179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(411..782)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /protein_id="ABS87289.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(411..782)
                     /gene="traJ"
                     /label=traJ
     oriT            815..924
                     /note="oriT"
     misc_feature    1056..1289
                     /note="similar to plasmid ColE1 in GenBank Accession Number
                     NC_001371"
     misc_feature    1281..1433
                     /note="similar to plasmid ColE1 in GenBank Accession Number
                     NC_001371"
     CDS             complement(1605..2255)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="ABS87290.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(1605..2255)
                     /gene="tetR"
                     /label=tetR
     CDS             2361..3560
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /protein_id="ABS87291.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            2361..3560
                     /gene="tetA"
                     /label=tetA
     CDS             complement(4191..4856)
                     /codon_start=1
                     /product="unknown plasmid factor 16.5"
                     /label=unknown plasmid factor 16.5
                     /protein_id="ABS87292.1"
                     /translation="MTWCIANARALWGQFRLGVQQPALYWHFRNKRALLDALASLSIAR
                     AHGALYELPINRGLKLTIVIKAQIRRLGAADVQDFREIRLSALKKAPEMFGSVYEHEEK
                     KPMEAFAERLRDAVAFGAYIDGEIIGLSVFKQEDGPKDAHKAHLSGVFVEPEQRGRGVA
                     GMLLRALPAGLLLVMIVRQIPTGIWWMRIFILGALNISLFWSLLFISVYRLPGGSRRR"
     CDS             complement(4831..5979)
                     /codon_start=1
                     /gene="trfA2"
                     /product="trans-acting replication A2"
                     /label=trfA2
                     /protein_id="ABS87293.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(4831..5979)
                     /gene="trfA2"
                     /label=trfA2
     CDS             complement(4831..5688)
                     /codon_start=1
                     /gene="trfA1"
                     /product="trans-acting replication A1"
                     /label=trfA1
                     /protein_id="ABS87294.1"
                     /translation="MTTKKRKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAP
                     WPDTMRAMPNDTARSALFTTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELV
                     WQQVLEYAKRTPIGEPITFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRV
                     GHLESVSLLHRFRVLDRGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRM
                     FDYFSSHREPYPLKLETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLV
                     HCKR"
     gene            complement(4831..5688)
                     /gene="trfA1"
                     /label=trfA1
     CDS             complement(6028..6378)
                     /codon_start=1
                     /gene="ssb"
                     /product="single-stranded DNA binding protein"
                     /label=ssb
                     /protein_id="ABS87295.1"
                     /translation="MSHNQFQFIGNLTRDTEVRHGNSNKPQAIFDIAVNEEWRNDAGDK
                     QERTDFFRIKCFGSQAEAHGKYLGKGSLVFVQGKIRNTKYEKDGQTVYGTDFIADKVDY
                     LDTKAPGGSNQE"
     gene            complement(6028..6378)
                     /gene="ssb"
                     /label=ssb
     CDS             6499..6864
                     /codon_start=1
                     /gene="trbA"
                     /product="TrbA"
                     /label=trbA
                     /protein_id="ABS87296.1"
                     /translation="MYNQIFFTNILRLLDERGMTKHELSERAGVSISFLSDLTNGKANP
                     SLKVMEAIADALETPLPLLLESTDLDREALAEIAGHPFKSSVPPGYERISVVLPSHKAF
                     IVKKWGDDTRKKLRGRL"
     gene            6499..6864
                     /gene="trbA"
                     /label=trbA
     CDS             complement(7902..8996)
                     /codon_start=1
                     /gene="incC1"
                     /product="IncC1"
                     /label=incC1
                     /protein_id="ABS87297.1"
                     /translation="MGVIHEETAYRKPVPGGDPGAGSGAADHRDSAGRLSRWEATGDVR
                     NVAGTDQGRSVASGASRVGRVRGQELARGVRAGNGGSAGTSGVHRPEVGSGRQEKTGNQ
                     TMKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNASYTLKDFATGLHAS
                     KLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDARELFGANIKALAN
                     QGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMVTTIANVRQKNAKL
                     QFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADALASGVPVWKIKKT
                     AARKASKEVRALADYVFTKMEISQ"
     gene            complement(7902..8996)
                     /gene="incC1"
                     /label=incC1
     CDS             complement(7902..8681)
                     /codon_start=1
                     /gene="incC2"
                     /product="IncC2"
                     /label=incC2
                     /protein_id="ABS87298.1"
                     /translation="MKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNAS
                     YTLKDFATGLHASKLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDA
                     RELFGANIKALANQGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMV
                     TTIANVRQKNAKLQFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADA
                     LASGVPVWKIKKTAARKASKEVRALADYVFTKMEISQ"
     gene            complement(7902..8681)
                     /gene="incC2"
                     /label=incC2
     CDS             complement(8678..8983)
                     /codon_start=1
                     /gene="korA"
                     /product="KorA"
                     /label=korA
                     /protein_id="ABS87299.1"
                     /translation="MKKRLTESQFQEAIQGLEVGQQTIEIARGVLVDGKPQATFATSLG
                     LTRGAVSQAVHRVWAAFEDKNLPEGYARVTAVLPEHQAYIVRKWEADAKKKQETKR"
     gene            complement(8678..8983)
                     /gene="korA"
                     /label=korA
     rep_origin      9448..10159
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      9723..10434
                     /note="oriV; incP"
     misc_feature    complement(10154..10307)
                     /note="similar to plasmid ColE1 in GenBank Accession Number
                     NC_001371"
     misc_feature    complement(10299..10529)
                     /note="similar to plasmid ColE1 in GenBank Accession Number
                     NC_001371"
     primer_bind     10669..10685
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10715..10739
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     10716..10736
                     /label=aatB1
                     /note="aatB1"
     RBS             10751..10771
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(10791..12062)
                     /codon_start=1
                     /gene="flp"
                     /product="Flp"
                     /label=flp
                     /note="FRT recognizing site-specific recombinase"
                     /protein_id="ABS87300.1"
                     /translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     gene            complement(10791..12062)
                     /gene="flp"
                     /label=flp
     RBS             12066..12074
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             12163..12876
                     /codon_start=1
                     /gene="cI"
                     /product="CI"
                     /label=cI
                     /note="regulatory protein"
                     /protein_id="ABS87301.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     gene            12163..12876
                     /gene="cI"
                     /label=cI
     protein_bind    complement(12894..14)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"