Basic Vector Information
YC3.6-N 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 ℃.
YC3.6-N vector Sequence
LOCUS 40924_49437 12837 bp DNA circular SYN 18-DEC-2018
DEFINITION Binary expression vector YC3.6-N, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12837)
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 12837)
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 12837)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12837)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2012"; volume: "69"; issue: "1"; pages: "181-192"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-OCT-2011) Department of Developmental Biology, Centre for
Organismal Studies Heidelberg, Im Neuenheimer Feld 230, Heidelberg
69120, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12837
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(114..138)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
protein_bind 303..324
/label=CAP binding site
/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 444..1089
/label=AtUBQ10
/note="AtUBQ10"
/regulatory_class="promoter"
CDS 1108..1791
/label=CFP
/note="cyan variant of GFP"
CDS 2248..2325
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 2548..3066
/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"
misc_feature 3075..3138
/label=multiple cloning site
/note="multiple cloning site"
terminator 3140..3781
/label=E9 terminator
/note="terminator and polyadenylation signal from the pea
rbcS-E9 gene"
primer_bind complement(3794..3810)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator complement(4392..4644)
/label=MAS terminator
/note="mannopine synthase terminator"
CDS complement(4843..5634)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
promoter complement(5664..6044)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
misc_feature complement(6416..6440)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 6968..7756
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
rep_origin 8003..8591
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(8777..8917)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(9261..9455)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9524..10588)
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(11025..11651)
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
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