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.
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pWHERE | Antibiotic Resistance | Ampicillin |
| Length | 9944 bp | Source | InvivoGen |
| Selection Marker | lacZ | Copy Number | High copy number |
| Cloning Method | Enzyme Cut | Expression Method | Transient |
pWHERE vector Vector Map
pWHERE vector Sequence
LOCUS Exported 9944 bp ds-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
FEATURES Location/Qualifiers
source 1..9944
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature complement(1..2032)
/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 2071..5157
/codon_start=1
/product="The E. coli lacZ gene codes for the enzyme
beta-galactosidase which catalyzes the hydrolysis of the
substrate X-Gal to produce a blue color that is easily
visualized under a microscope. A nuclear localization
signal of SV40 large T has been inserted in the 5' end of
the lacZ gene to allow the targeting of the chimeric
protein to the nucleus. To reduce the immunogenicity of
this bacterial gene, InvivoGen has engineered a synthetic
lacZnls gene that is entirely free of CpG motifs, whereas
the wild type lacZ gene contains 298 CpG dinucleotides."
/note="LacZ Delta-CpG NLS"
/translation="MDPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQ
LRSLNGEWRFAWFPAPEAVPESWLECDLPEAVPKKKRKVEADTVVVPSNWQMHGYDAPI
YTNVTYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRW
VGYGQDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHK
PTTQISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGG
EIIDERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREV
RIENGLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPN
HPLWYTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSV
IIWSLGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPF
PAVPKWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWV
DQSLIKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFR
LSGQTIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPE
SAGQLWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFC
IELGNKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWV
ERWKAAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAI
TVDVEVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDM
YTPYVFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWL
NIDGFHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
CDS 2302..2322
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
polyA_signal 5170..5742
/note="EF-1-alpha poly(A) signal"
/note="human EF-1-alpha polyadenylation signal"
misc_feature complement(5750..7781)
/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
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8636..9496)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(9497..9601)
/gene="bla"
/note="AmpR promoter"
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