Basic Vector Information
- Vector Name:
- PC-GW-Kan
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 11797 bp
- Type:
- Binary vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Dalal J, Yalamanchili R, La Hovary C, Ji M, Rodriguez-Welsh M, Aslett D, Ganapathy S, Grunden A, Sederoff H, Qu R.
- Promoter:
- CaMV 35S (enhanced)
PC-GW-Kan vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
PC-GW-Kan vector Sequence
LOCUS 40924_7771 11797 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector PC-GW-Kan, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11797)
AUTHORS Dalal J, Yalamanchili R, La Hovary C, Ji M, Rodriguez-Welsh M,
Aslett D, Ganapathy S, Grunden A, Sederoff H, Qu R.
TITLE A novel gateway-compatible binary vector series (PC-GW) for flexible
cloning of multiple genes for genetic transformation of plants
JOURNAL Plasmid 81, 55-62 (2015)
PUBMED 26188330
REFERENCE 2 (bases 1 to 11797)
AUTHORS Dalal J, Yalamanchili R, Hovary CL, Ji M, Rodriguez-Welsh M, Aslett
D, Ganapathy S, Grunden A, Sederoff H, Qu R.
TITLE Direct Submission
JOURNAL Submitted (19-FEB-2015) Crop Science, North Carolina State
University, Campus Box 7287, Raleigh, NC 27695, USA
REFERENCE 3 (bases 1 to 11797)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11797)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 81,
55-62 (2015)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-FEB-2015) Crop Science, North Carolina State University, Campus
Box 7287, Raleigh, NC 27695, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11797
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1219..1845
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 2282..3346
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 3415..3609
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 3953..4093
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4279..4867)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4957..5748)
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 6173..6197
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(6275..6449)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6509..7297)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
promoter complement(7366..8043)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 8234..8255
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8270..8300
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8308..8324
/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 8332..8348
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 8965..9310
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 9399..9518
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 9548..9578
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 9632..10288
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 10633..10935
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(11010..11134)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
regulatory 11238..11463
/label=35S terminator from pSIM1 cloning vector
/note="35S terminator from pSIM1 cloning vector"
/regulatory_class="terminator"
primer_bind complement(11473..11489)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 11692..11716
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
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