LOCUS       Exported                7567 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5333, complete sequence.
ACCESSION   EU024543
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7567)
  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 7567)
  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 7567)
  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 7567)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7567
                     /organism="Cloning vector pCPP5333"
                     /mol_type="other DNA"
                     /db_xref="taxon:458223"
     misc_feature    14..1181
                     /label=pUC18 fragment
                     /note="pUC18 fragment"
     primer_bind     complement(29..45)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    53..69
                     /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(77..107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    122..143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      431..1019
                     /label=oriV
                     /note="oriV"
     CDS             complement(1272..1805)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1"
                     /label=aacC1
                     /note="gentamicin acetyltransferase"
                     /protein_id="ABS87302.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(1272..1805)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(1994..2022)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(2040..2144)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2234..2251
                     /label=SCE
                     /note="SCE"
     CDS             complement(2473..3894)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="levan sucrase precursor"
                     /protein_id="ABS87303.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(2473..3894)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(3895..4340)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     oriT            4368..5192
                     /note="oriT"
     oriT            4714..4823
                     /note="incP origin of transfer"
     terminator      5244..5271
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      complement(5244..5270)
                     /regulatory_class="terminator"
                     /note="T2"
     terminator      5363..5449
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      complement(5403..5446)
                     /regulatory_class="terminator"
                     /note="T1"
     primer_bind     5799..5815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5860..7564
                     /label=Gateway cassette RfA
                     /note="Gateway cassette RfA"
     misc_recomb     5860..5984
                     /label=attR1
                     /note="attR1"
     protein_bind    5860..5984
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6009..6039
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6093..6752
                     /codon_start=1
                     /gene="CmR"
                     /product="CAT"
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /protein_id="ABS87304.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            6093..6752
                     /gene="CmR"
                     /label=CmR
     CDS             7094..7399
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="gyrase toxin protein"
                     /protein_id="ABS87305.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            7094..7399
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     complement(7440..7564)
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(7440..7564)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"