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pB1H2w2-zif268 vector (V012168)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012168 pB1H2w2-zif268 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

Vector Name:
pB1H2w2-zif268
Antibiotic Resistance:
Ampicillin
Length:
3966 bp
Type:
Bacterial Expression
Replication origin:
ori
Copy Number:
High Copy
Cloning Method:
Restriction Enzyme
5' Primer:
OMG5

pB1H2w2-zif268 vector Map

Copy Sequence View Map
pB1H2w2-zif2683966 bp60012001800240030003600lac operatorRNAP omegaFLAGEarly growth response protein 1f1 oriAmpR promoterAmpRoriL4440bomrop

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.

pB1H2w2-zif268 vector Sequence

LOCUS       V012168                 3966 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V012168
VERSION     V012168
KEYWORDS    pB1H2w2-zif268
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 3966)
  AUTHORS   Noyes MB, Meng X, Wakabayashi A, Sinha S, Brodsky MH, Wolfe SA
  TITLE     A systematic characterization of factors that regulate Drosophila
            segmentation via a bacterial one-hybrid system.
  JOURNAL   Nucleic Acids Res. 2008 Mar 10. ():.
   PUBMED   18332042
REFERENCE   2  (bases 1 to 3966)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3966)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res. 2008 Mar 10. ():."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    50..66
                     /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)."
     CDS             306..575
                     /label="RNAP omega"
                     /note="DNA-directed RNA polymerase subunit omega"
     CDS             585..608
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             672..848
                     /gene="EGR1"
                     /label="Early growth response protein 1"
                     /note="Early growth response protein 1 from Serinus
                     canaria. Accession#: O73694"
     rep_origin      1027..1482
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1508..1612
                     /label="AmpR promoter"
     CDS             1613..2470
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2644..3232
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     3386..3403
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     misc_feature    complement(3418..3558)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     primer_bind     3644..3666
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3663..3851)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"