Basic Vector Information
- Vector Name:
- pVC4-bar
- Antibiotic Resistance:
- Kanamycin
- Length:
- 11224 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Conn SJ, Conn V, Tyerman SD, Kaiser BN, Leigh RA, Gilliham M.
- Promoter:
- CaMV 35S (enhanced)
pVC4-bar 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 ℃.
pVC4-bar vector Sequence
LOCUS 40924_45898 11224 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pVC4-bar, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11224)
AUTHORS Conn SJ, Conn V, Tyerman SD, Kaiser BN, Leigh RA, Gilliham M.
TITLE Magnesium transporters, MGT2/MRS2-1 and MGT3/MRS2-5, are important
for magnesium partitioning within Arabidopsis thaliana mesophyll
vacuoles
JOURNAL New Phytol. 190 (3), 583-594 (2011)
PUBMED 21261624
REFERENCE 2 (bases 1 to 11224)
AUTHORS Conn V.
TITLE Direct Submission
JOURNAL
REFERENCE 3 (bases 1 to 11224)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11224)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "New
Phytol."; date: "2011"; volume: "190"; issue: "3"; pages: "583-594"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-AUG-2010) Agriculture, Food "
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11224
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(82..106)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
promoter 526..870
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 1078..1094
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1175..1299
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1324..1354
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1408..2064
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2409..2711
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2755..2879)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 2912..3159
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3191..3207)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3215..3231
/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(3239..3269)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3284..3305)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3496..4173
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 4218..4766
/codon_start=1
/label=BlpR
/note="phosphinothricin acetyltransferase"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
polyA_signal 4776..4950
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature complement(5028..5052)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 5477..6268
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 6358..6946
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(7132..7272)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7616..7810)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(7879..8943)
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
CDS complement(9380..10006)
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
This page is informational only.