Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | D3cpv-C | Antibiotic Resistance | Spectinomycin/Streptomycin |
| Length | 12828 bp | Type | Binary expression vector |
| Source | Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, Kudla J, Schumacher K. | ||
D3cpv-C 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 ℃.
D3cpv-C vector Sequence
LOCUS Exported 12828 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Binary expression vector D3cpv-C, complete sequence.
ACCESSION JN903571
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12828)
AUTHORS Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M,
Kudla J, Schumacher K.
TITLE FRET-based genetically encoded sensors allow high-resolution live
cell imaging of Ca(2) dynamics
JOURNAL Plant J. 69 (1), 181-192 (2012)
PUBMED 21910770
REFERENCE 2 (bases 1 to 12828)
AUTHORS Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M,
Kudla J, Schumacher K.
TITLE Direct Submission
JOURNAL Submitted (24-OCT-2011) Department of Developmental Biology, Centre
for Organismal Studies Heidelberg, Im Neuenheimer Feld 230,
Heidelberg 69120, Germany
REFERENCE 3 (bases 1 to 12828)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12828
/organism="Binary expression vector D3cpv-C"
/mol_type="other DNA"
/db_xref="taxon:1154701"
misc_feature 1..126
/label=right border
/note="right border"
misc_feature 114..138
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
protein_bind 303..324
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 339..369
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 377..393
/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 401..417
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 443..1047
/regulatory_class="promoter"
/note="AtUBQ10"
misc_feature 1077..1130
/label=multiple cloning site
/note="multiple cloning site"
regulatory 1148..1157
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1154..3115
/codon_start=1
/product="D3cpv"
/label=D3cpv
/protein_id="AFC41192.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAARMHDQLTEEQIAEFKEAFSLLDKDGDGTITTKELGTALRSLGQNPTEAELQDM
INEVDADGNGTIYFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMT
NLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAKGGKRRWQKTGHAVRAFGRLKK
ISSSGALELMDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGD
ATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQE
RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA
DKQKNGIKANFKIRHNIE"
CDS 2594..3112
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIE"
regulatory 3130..3776
/regulatory_class="terminator"
/note="rbcs"
terminator 3131..3772
/label=E9 terminator
/note="terminator and polyadenylation signal from the pea
rbcS-E9 gene"
primer_bind complement(3785..3801)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory complement(3965..4657)
/regulatory_class="terminator"
/note="mas"
terminator 4383..4635
/label=MAS terminator
/note="mannopine synthase terminator"
CDS complement(4831..5625)
/codon_start=1
/product="nptII"
/label=nptII
/protein_id="AFC41193.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(5655..6035)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
regulatory complement(5656..6340)
/regulatory_class="promoter"
/note="mas"
misc_feature 6341..6667
/label=left border
/note="left border"
misc_feature 6407..6431
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 6818..7750
/codon_start=1
/product="aadA"
/label=aadA
/protein_id="AFC41194.1"
/translation="MTCFFGVQSMPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKL
NIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVR
LDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRND
ILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNS
PPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQ
EEDRLASRADQLEEFVHYVKGEITKVVGK"
CDS 6959..7750
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 7994..8582
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 8768..8908
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 9252..9446
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9512..10579)
/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"
CDS complement(11013..11642)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
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