pWHERE vector (V012687)

Basic Vector Information

The pWHERE plasmid was designed for studies of temporal expression and tissue distribution of your promoter of interest, cloned within an insulated LacZ cassette, in transgenic mice and rats.

      • Vector Name:
      • pWHERE
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9944 bp
      • Source/Author:
      • InvivoGen
      • Selection Marker:
      • lacZ
      • Copy Number:
      • High copy number
      • Cloning Method:
      • Enzyme Cut
      • Expression Method:
      • Transient

pWHERE vector Vector Map

pWHERE9944 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600H19 insSV40 NLSEF-1-alpha poly(A) signalH19 insoriAmpRAmpR 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 ℃.

pWHERE vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pWHERE.        9944 bp DNA     circular SYN 06-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pWHERE.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9944)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9944)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..2032)
                     /label=H19 ins
                     /note="H19 ins"
                     /note="mH19 insulators on either side of the lacZ
                     transcription unit. Both insulators are expected to protect
                     the integrated transcriptional LacZ unit from negative as 
                     well as positive influences from neighboring sequences. 
                     Insulator elements can be functionally identified by their 
                     ability to shield promoters from regulators in a 
                     position-dependent manner or by their ability to protect 
                     adjacent transgenes from position effects. The fragment of 
                     the differential methylated region (DMD) located between 
                     the mouse Igf2 and H19 acts as a powerful insulator. The 
                     enhancer blocking activity of the DMD fragment is dependent
                     upon four responsive elements to the vertebrate 
                     enhancer-blocking protein CTCF. Two mouse DMD fragments 
                     have been introduced in opposite orientation in the pWHERE 
                     plasmid to insulate your promoter of interest cloned 
                     upstream of the new CpG-free LacZ gene from the 5' and 3' 
                     adjacent regions at the integrated site in transgenic 
                     mice."
     CDS             2302..2322
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    5170..5742
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     misc_feature    complement(5750..7781)
                     /label=H19 ins
                     /note="H19 ins"
                     /note="mH19 insulators on either side of the lacZ
                     transcription unit. Both insulators are expected to protect
                     the integrated transcriptional LacZ unit from negative as 
                     well as positive influences from neighboring sequences. 
                     Insulator elements can be functionally identified by their 
                     ability to shield promoters from regulators in a 
                     position-dependent manner or by their ability to protect 
                     adjacent transgenes from position effects. The fragment of 
                     the differential methylated region (DMD) located between 
                     the mouse Igf2 and H19 acts as a powerful insulator. The 
                     enhancer blocking activity of the DMD fragment is dependent
                     upon four responsive elements to the vertebrate 
                     enhancer-blocking protein CTCF. Two mouse DMD fragments 
                     have been introduced in opposite orientation in the pWHERE 
                     plasmid to insulate your promoter of interest cloned 
                     upstream of the new CpG-free LacZ gene from the 5' and 3' 
                     adjacent regions at the integrated site in transgenic 
                     mice."
     rep_origin      complement(7877..8465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8639..9496)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9497..9601)
                     /label=AmpR promoter

This page is informational only.