pCPP5333 vector (V008216)

Basic Vector Information

Vector Name:
pCPP5333
Antibiotic Resistance:
Chloramphenicol
Length:
7567 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Wei CF, Kvitko BH, Shimizu R, Crabill E, Alfano JR, Lin NC, Martin GB, Huang HC, Collmer A.
Promoter:
sacB

pCPP5333 vector Map

pCPP53337567 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500M13 revlac operatorlac promoterCAP binding siteoriGmRPc promoterAmpR promoterSCESacBsacB promoteroriTrrnB T2 terminatorrrnB T1 terminatorM13 fwdattR1lac UV5 promoterCmRccdBattR2

pCPP5333 vector Sequence

LOCUS       40924_13160        7567 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5333, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7567)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J."; 
            date: "2007"; volume: "51"; issue: "1"; pages: "32-46"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "2007"; volume: "189"; issue: "22"; pages: 
            "8059-8072"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (09-JUL-2007) Plant Pathology, Cornell University, 334 Plant 
            Science, Ithaca, NY 14853, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..45)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(53..69)
                     /label=lac operator
                     /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    complement(122..143)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(431..1019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1275..1805)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1994..2022)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(2040..2144)
                     /label=AmpR promoter
     misc_feature    2234..2251
                     /label=SCE
                     /note="SCE"
     CDS             complement(2476..3894)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(3895..4340)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     oriT            complement(4714..4823)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(5244..5271)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5363..5449)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     5799..5815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5860..5979
                     /label=attR1
                     /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..6749
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             7094..7396
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(7440..7564)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"

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