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PC-GW-EGFP vector (V008973#)

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

pVS1 StaA pVS1 RepA pVS1 oriV bom ori aadA left border T-DNA repeat CaMV poly(A) signal EGFP CamV 35S2 promoter CAP binding site lac promoter lac operator M13 rev PC-GW M13 fwd RB T-DNA repeat

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

Copy Sequence

Download GeneBank File(.gb)

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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