Basic Vector Information
- Vector Name:
- pENTR4
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 3757 bp
- Type:
- Gateway Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- Promoter:
- lac UV5
pENTR4 vector Vector Map
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 ℃.
pENTR4 vector Sequence
LOCUS pENTR4. 3757 bp DNA circular SYN 01-JAN-1980
DEFINITION Gateway(R) Dual Selection Vector for creating an entry vector by
restriction cloning. Same MCS as pENTR(TM)1A except with an NcoI
site.
ACCESSION .
VERSION .
KEYWORDS pENTR4.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3757)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3757)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT To create a Kozak sequence, clone a blunt fragment starting with
ATGG (where ATG is the start codon) into the XmnI site.
FEATURES Location/Qualifiers
source 1..3757
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 103..189
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..457
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 461..510
/label=MCS 1
/note="MCS 1"
/note="multiple cloning site, part 1"
promoter 527..557
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 611..1288
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 1611..1913
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
misc_feature 1951..1982
/label=MCS 2
/note="MCS 2"
/note="multiple cloning site, part 2"
protein_bind complement(1986..2085)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2208..3014
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 3107..3695
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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