pBIND vector (V000416)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V000416	pBIND	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

The pBIND Vector is a high-copy plasmid in which the CMV immediate early promoter drives expression of a portion of the yeast GAL4 gene (amino acids 1–147) containing a DNA-binding domain. As with the pACT Vector, the DNAbinding domain sequence is flanked by a chimeric intron, a multiple cloning region, stop codons and an SV40 late polyadenylation region (Figures 4 and 5). The fusion gene region is flanked by T7 and T3 RNA polymerase promoters for the purpose of synthesizing sense and antisense RNA products, respectively. The Renilla luciferase gene on this vector is preceded by the SV40 early promoter and a growth hormone intron. Introns can increase protein expression through mRNA stability and nuclear to cytoplasmic transport effects (9–12). A synthetic polyadenylation sequence resides 3´ of the Renilla luciferase gene. The plasmid backbone contains an f1 origin of replication for the production of ssDNA and the β-lactamase gene (Ampr) for selection in E. coli.

Vector Name:: pBIND

Antibiotic Resistance:: Ampicillin

Length:: 6360 bp

Type:: Mammalian Two-Hybrid System

Replication origin:: ori

Promoter:: CMV

pBIND vector Map

Copy Sequence View Map

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pBIND vector Sequence

LOCUS       40924_6527        6360 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6360)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1116..1556
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
     promoter        complement(1640..1657)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    complement(1675..1796)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1982..2437
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2509..2866
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3208..4140
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
     polyA_signal    4201..4249
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        4563..4667
                     /label=AmpR promoter
     CDS             4668..5525
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5699..6287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"