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pUC-triRV vector (V002437#)

Basic Vector Information

Basic Vector Information
Vector Name pUC-triRV Antibiotic Resistance Ampicillin
Length 4199 bp Type BiFC expression vector
Source Offenborn JN, Waadt R, Kudla J.

pUC-triRV vector Vector Map

AmpR promoter AmpR pBR322 origin CAP binding site lac promoter lac operator M13 rev p35S triRV NosT M13 fwd

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 ℃.

pUC-triRV vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4199 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  BiFC expression vector pUC-triRV, complete sequence.
ACCESSION   KR857706
VERSION     .
KEYWORDS    triFC expression vector.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4199)
  AUTHORS   Offenborn JN, Waadt R, Kudla J.
  TITLE     Visualization and translocation of ternary 
            Calcineurin-A/Calcineurin-B/Calmodulin-2 protein complexes by 
            dual-color trimolecular fluorescence complementation
  JOURNAL   New Phytol. (2015) In press
  PUBMED    25919910
REFERENCE   2  (bases 1 to 4199)
  AUTHORS   Offenborn JN, Waadt R, Kudla J.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2015) University of Muenster, Institute for 
            Biology and Plant Biotechnology, Schlossplatz 7, Muenster 48149, 
            Germany
REFERENCE   3  (bases 1 to 4199)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4199
                     /organism="BiFC expression vector pUC-triRV"
                     /mol_type="other DNA"
                     /db_xref="taxon:1662180"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      131..159
                     /regulatory_class="promoter"
                     /note="amp prom"
     CDS             201..1061
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AKJ78219.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1216..1835
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      2144..2173
                     /regulatory_class="promoter"
                     /note="lac prom"
     protein_bind    2182..2198
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2241..3104
                     /label=p35S
                     /note="p35S"
     promoter        2751..3096
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3111..3707
                     /codon_start=1
                     /product="triRV"
                     /label=triRV
                     /protein_id="AKJ78218.1"
                     /translation="MGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIK
                     LDITSHNEDYTIVEQYERAEGRHSTGGMDELYKYPYDVPDYAGPRPTSGSVDLEGTAPG
                     MYPYDVPDYADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSY
                     QSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     misc_feature    3114..3344
                     /label=mCherry 160-236
                     /note="mCherry 160-236"
     misc_feature    3345..3371
                     /label=HA-Tag
                     /note="HA-Tag"
     CDS             3345..3371
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3372..3422
                     /label=MCS2
                     /note="MCS2"
     misc_feature    3426..3452
                     /label=HA-Tag
                     /note="HA-Tag"
     CDS             3426..3452
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3453..3704
                     /label=mVenus 156-239
                     /note="mVenus 156-239"
     CDS             3453..3704
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     misc_feature    3714..3978
                     /label=NosT
                     /note="NosT"
     terminator      3724..3976
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3985..4001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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