Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | PC-GW-EGFP | Antibiotic Resistance | Chloramphenicol |
| Length | 11645 bp | Type | Binary vector |
| Source | Dalal J, Yalamanchili R, La Hovary C, Ji M, Rodriguez-Welsh M, Aslett D, Ganapathy S, Grunden A, Sederoff H, Qu R. | ||
PC-GW-EGFP 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-EGFP vector Sequence
LOCUS Exported 11645 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Binary vector PC-GW-EGFP, complete sequence.
ACCESSION KP826772
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11645)
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 11645)
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 11645)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11645
/organism="Binary vector PC-GW-EGFP"
/mol_type="other DNA"
/db_xref="taxon:1642502"
CDS 1219..1848
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2282..3349
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
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(4954..5748)
/codon_start=1
/gene="aadA"
/product="kanamycin resistance protein"
/label=aadA
/note="AadA"
/protein_id="AKC88580.1"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
gene complement(4954..5748)
/gene="aadA"
/label=aadA
misc_feature 6173..6198
/label=left border T-DNA repeat
/note="left border T-DNA repeat"
misc_feature 6173..6197
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 6275..6449
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6456..7175)
/codon_start=1
/gene="EGFP"
/product="enhanced green fluorescent protein"
/label=EGFP
/protein_id="AKC88579.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
gene complement(6456..7175)
/gene="EGFP"
/label=EGFP
regulatory complement(7182..7962)
/regulatory_class="promoter"
/note="CamV 35S2 promoter"
promoter complement(7214..7891)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 8082..8103
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8118..8148
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8156..8172
/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 8180..8196
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 8211..11312
/label=PC-GW
/note="PC-GW"
regulatory 8326..9161
/regulatory_class="promoter"
/note="35S promoter from PUGW5 cloning vector"
promoter 8813..9158
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 9184..10982
/label=Gateway region from pEarleyGate100
/note="Gateway region from pEarleyGate100"
misc_recomb 9184..9366
/label=attR1 from pEarleyGate100
/note="attR1 from pEarleyGate100"
protein_bind 9247..9371
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 9396..9426
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 9480..10139
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPVSRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 10481..10786
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/protein_id="AKC88581.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 10481..10786
/gene="ccdB"
/label=ccdB
misc_recomb 10858..10982
/label=attR2 from pEarleyGate100
/note="attR2 from pEarleyGate100"
protein_bind complement(10858..10982)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 11086..11311
/regulatory_class="terminator"
/note="35S terminator from pSIM1 cloning vector"
primer_bind complement(11321..11337)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 11540..11564
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
misc_feature 11559..11584
/label=right border T-DNA repeat from pCAMBIA 2300
/note="right border T-DNA repeat from pCAMBIA 2300"
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