Basic Vector Information
- Vector Name:
- pMDC32-HPB
- Antibiotic Resistance:
- Kanamycin
- Length:
- 12097 bp
- Type:
- Gateway destination vector
- Replication origin:
- ori
- Source/Author:
- Qi Y, Katagiri F.
- Promoter:
- CaMV35S(enhanced)
pMDC32-HPB 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 ℃.
pMDC32-HPB vector Sequence
LOCUS 40924_30435 12097 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway destination vector pMDC32-HPB, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12097)
AUTHORS Qi Y, Katagiri F.
TITLE Purification of low-abundance Arabidopsis plasma-membrane protein
complexes and identification of candidate components
JOURNAL Plant J. 57 (5), 932-944 (2009)
PUBMED 19000159
REFERENCE 2 (bases 1 to 12097)
AUTHORS Qi Y, Katagiri F.
TITLE Direct Submission
JOURNAL Submitted (29-AUG-2008) Plant Biology, University of Minnesota, 1500
Gortner Ave., St.Paul, MN 55414, USA
REFERENCE 3 (bases 1 to 12097)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12097)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2009"; volume: "57"; issue: "5"; pages: "932-944"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-AUG-2008) Plant Biology, University of Minnesota, 1500 Gortner
Ave., St.Paul, MN 55414, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12097
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(6..253)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(576..599)
/codon_start=1
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
/translation="LEVLFQGP"
CDS complement(609..635)
/codon_start=1
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
protein_bind 664..788
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(832..1134)
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1479..2135)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(2189..2219)
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind complement(2244..2368)
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(2428..2772)
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind complement(3199..3215)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3418..3442
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4742..5368
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 5805..6869
/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"
rep_origin 6938..7132
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7476..7616
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7802..8390)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8480..9271)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 9696..9720
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(9798..9972)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(10015..11037)
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(11105..11782)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 11973..11994
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 12009..12039
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12047..12063
/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 12071..12087
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
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