LOCUS       Exported                8791 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5702, complete sequence.
ACCESSION   EU024546
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8791)
  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 8791)
  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 8791)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8791
                     /organism="Cloning vector pCPP5702"
                     /mol_type="other DNA"
                     /db_xref="taxon:458225"
     rep_origin      46..397
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             411..1244
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /protein_id="ABS87320.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            411..1244
                     /gene="rep"
                     /label=rep
     primer_bind     1405..1421
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1425..1548
                     /regulatory_class="terminator"
                     /note="omega"
     CDS             complement(2033..2827)
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /label=nptII
                     /note="kanamycin resistance protein"
                     /protein_id="ABS87321.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2033..2827)
                     /gene="nptII"
                     /label=nptII
     regulatory      complement(3341..3464)
                     /regulatory_class="terminator"
                     /note="omega"
     regulatory      3465..3497
                     /regulatory_class="promoter"
                     /note="hrp box"
     CDS             3532..5289
                     /codon_start=1
                     /gene="avrPto1-cya"
                     /product="AvrPto1-Cya fusion protein"
                     /label=avrPto1-cya
                     /note="AvrPto1 from P. syringae pv.tomato DC3000 fused to 
                     the catalytic domain of calmodulin-dependant adenylate 
                     cyclase from Bordatella pertussis"
                     /protein_id="ABS87322.1"
                     /translation="MGNICVGGSRMAHQVNSPDRVSNNSGDEDNVTSSQLLSVRHQLAE
                     SAGVPRDQHEFVSNQAPQSLRNRYNNLYSHTQRTLDMADMQHRYMTGASGINPGMLPHE
                     NVDDMRSAITDWSDMREALQYAMGIHADIPPSPERFVATMNPNGSIRMSTLSPSPYRNW
                     QKGGRADPAFLYKVVDGQQSHQAGYANAADRESGIPAAVLDGIKAVAKEKNATLMFRLV
                     NPHSTSLIAEGVATKGLGVHAKSSDWGLQAGYIPVNPNLSKLFGRAPEVIARADNDVNS
                     SLAHGHTAVDLTLSKERLDYLRQAGLVTGMADGVVASNHAGYEQFEFRVKETSDGRYAV
                     QYRRKGGDDFEAVKVIGNAAGIPLTADIDMFAIMPHLSNFRDSARSSVTSGDSVTDYLA
                     RTRRAASEATGGLDRERIDLLWKIARAGARSAVGTEARRQFRYDGDMNIGVITDFELEV
                     RNALNRRAHAVGAQDVVQHGTEQNNPFPEADEKIFVVSATGESQMLTRGQLKEYIGQQR
                     GEGYVFYENRAYGVAGKSLFDDGLGAAPGVPSGRSKFSPDVLETVPASPGLRRPSLGAV
                     ERQDSGYDS"
     gene            3532..5289
                     /gene="avrPto1-cya"
                     /label=avrPto1-cya
     misc_feature    4024..4068
                     /gene="avrPto1-cya"
                     /label=linker
                     /note="linker"
     protein_bind    complement(4040..4064)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5305..5321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5329..5345
                     /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(5353..5383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5398..5419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5707..6295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7117..8307)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracyclin efflux protein"
                     /label=tet
                     /protein_id="ABS87323.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(7117..8307)
                     /gene="tet"
                     /label=tet
     promoter        complement(8355..8383)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     protein_bind    8630..8651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8666..8696
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8704..8720
                     /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)."
     primer_bind     8728..8744
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8762..8780
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"