PC-GW-Hyg vector (V008972)

Basic Vector Information

      • Vector Name:
      • PC-GW-Hyg
      • Antibiotic Resistance:
      • Chloramphenicol
      • Length:
      • 12013 bp
      • Type:
      • Binary vector
      • Source/Author:
      • Dalal J, Yalamanchili R, La Hovary C, Ji M, Rodriguez-Welsh M, Aslett D, Ganapathy S, Grunden A, Sederoff H, Qu R.

PC-GW-Hyg vector Vector Map

PC-GW-Hyg12013 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000PC-GWM13 fwdRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriaadAleft border T-DNA repeatCaMV poly(A) signalHPTIICamV 35S2 promoterCAP binding sitelac promoterlac operatorM13 rev

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               12013 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector PC-GW-Hyg, complete sequence.
ACCESSION   KP826770
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12013
                     /organism="Binary vector PC-GW-Hyg"
                     /mol_type="other DNA"
                     /db_xref="taxon:1642503"
     misc_feature    7..3106
                     /label=PC-GW
                     /note="PC-GW"
     regulatory      121..956
                     /regulatory_class="promoter"
                     /note="35S promoter from PUGW5 cloning vector"
     promoter        608..953
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    979..2777
                     /label=Gateway region from pEarleyGate100
                     /note="Gateway region from pEarleyGate100"
     misc_recomb     979..1161
                     /label=attR1 recombination site from pEarleyGate100
                     /note="attR1 recombination site from pEarleyGate100"
     protein_bind    1042..1166
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /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..1934
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPVSRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2276..2581
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="AKC88575.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2276..2581
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     2653..2777
                     /label=attR2 recombination site from pEarleyGate100
                     /note="attR2 recombination site from pEarleyGate100"
     protein_bind    complement(2653..2777)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2881..3106
                     /regulatory_class="terminator"
                     /note="35S terminator from pSIM1 cloning vector"
     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"
     misc_feature    3354..3379
                     /label=right border T-DNA repeat from pCAMBIA 2300
                     /note="right border T-DNA repeat from pCAMBIA 2300"
     CDS             4659..5288
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5722..6789
                     /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      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(8394..9188)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin resistance protein"
                     /label=aadA
                     /note="AadA"
                     /protein_id="AKC88573.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(8394..9188)
                     /gene="aadA"
                     /label=aadA
     misc_feature    9613..9638
                     /label=left border T-DNA repeat
                     /note="left border T-DNA repeat"
     misc_feature    9613..9637
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9715..9889
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9929..10954)
                     /codon_start=1
                     /gene="HPTII"
                     /product="hygromycin phosphotransferase protein"
                     /label=HPTII
                     /protein_id="AKC88574.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(9929..10954)
                     /gene="HPTII"
                     /label=HPTII
     regulatory      complement(10990..11770)
                     /regulatory_class="promoter"
                     /note="CamV 35S2 promoter"
     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
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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     11988..12004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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