Basic Vector Information
- Vector Name:
- PC-GW-Hyg
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 12013 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-Hyg 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-Hyg vector Sequence
LOCUS 40924_7766 12013 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector PC-GW-Hyg, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12013)
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 12013)
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 12013)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12013)
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..12013
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 608..953
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 1042..1161
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1191..1221
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1275..1931
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 2276..2578
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(2653..2777)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
regulatory 2881..3106
/label=35S terminator from pSIM1 cloning vector
/note="35S terminator from pSIM1 cloning vector"
/regulatory_class="terminator"
primer_bind complement(3116..3132)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3335..3359
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4659..5285
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 5722..6786
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 6855..7049
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7393..7533
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7719..8307)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8397..9188)
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 9613..9637
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(9715..9889)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9932..10954)
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
promoter complement(11022..11699)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 11890..11911
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11926..11956
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11964..11980
/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)."
primer_bind 11988..12004
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
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