FRB-NLuc vector (V001009)

Basic Vector Information

      • Vector Name:
      • FRB-NLuc
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7145 bp
      • Type:
      • Mammalian Expression
      • Replication origin:
      • ori
      • Selection Marker:
      • Neomycin (select with G418)
      • Copy Number:
      • High Copy
      • Cloning Method:
      • TOPO Cloning

FRB-NLuc vector Vector Map

FRB-NLuc7145 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900pRS-markerCMV enhancerCMV promoterT7 promoterFRBLucNrevV5 tag6xHisbGH poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoter

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 ℃.

FRB-NLuc vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_865        7145 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  AUTHORS   Luker KE, Smith MC, Luker GD, Gammon ST, Piwnica-Worms H, 
            Piwnica-Worms D
  TITLE     Kinetics of regulated protein-protein interactions revealed with 
            firefly luciferase complementation imaging in cells and living 
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Aug 17. 101(33):12288-93.
  PUBMED    15284440
REFERENCE   2  (bases 1 to 7145)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7145)
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2004 Aug 17. 101(33):12288-93."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /note="pRS vectors, use to sequence yeast selectable
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             965..1234
                     /note="FKBP-rapamycin binding domain of human FRAP"
     primer_bind     complement(1343..1361)
                     /note="Firefly luciferase, reverse primer"
     CDS             2642..2683
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             2693..2710
                     /note="6xHis affinity tag"
     polyA_signal    2739..2963
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3009..3437
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3451..3781
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3848..4639
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4818..4951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
     protein_bind    complement(5012..5028)
                     /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)."
     promoter        complement(5036..5066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5081..5102)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(5219..5236)
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5390..5978)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(6152..7009)
     promoter        complement(7010..7114)
                     /label=AmpR promoter

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