Basic Vector Information
- Vector Name:
- pEG BacMam N term StrepII eGFP 3C
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7695 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Insect cells
- Promoter:
- CMV
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pEG BacMam N term StrepII eGFP 3C 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 ℃.
pEG BacMam N term StrepII eGFP 3C vector Sequence
LOCUS 62056_8910 7695 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 7695)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7695
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 340..472
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
CDS 541..564
/codon_start=1
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
/translation="WSHPQFEK"
CDS 574..1287
/codon_start=1
/label=mEGFP
/note="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS 1306..1329
/codon_start=1
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
/translation="LEVLFQGP"
misc_feature 1407..1995
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal 2128..2262
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
mobile_element complement(2291..2456)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin 2640..3094
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3120..3224
/label=AmpR promoter
CDS 3225..4082
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4256..4844
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
mobile_element complement(5149..5373)
/label=Tn7R
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(5443..5973)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(6162..6190)
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal complement(6707..6755)
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
promoter complement(6891..7000)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 7045..7136
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
enhancer 7317..7695
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
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