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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