Basic Vector Information
- Vector Name:
- pEHlyA5
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3990 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- E. coli
- Promoter:
- T7
- 5' Primer:
- M13F
- Growth Temperature:
- 37℃
pEHlyA5 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 ℃.
pEHlyA5 vector Sequence
LOCUS 62056_9120 3990 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3990)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3990
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 510..531
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 546..576
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 584..600
/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)."
primer_bind 608..624
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 666..684
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 716..738
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 752..769
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 1184..1210
/codon_start=1
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
CDS 1211..1234
/codon_start=1
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
/translation="LEVLFQGP"
primer_bind complement(1998..2014)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2488..2592
/label=AmpR promoter
CDS 2593..3450
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3624..3990
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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