Basic Vector Information
- Vector Name:
- pENN.AAV.cTNT.PI.eGFP.WPRE.rBG
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5573 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Adeno-associated virus
- Promoter:
- cTnT
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pENN.AAV.cTNT.PI.eGFP.WPRE.rBG 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 ℃.
pENN.AAV.cTNT.PI.eGFP.WPRE.rBG vector Sequence
LOCUS 62056_9445 5573 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 5573)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5573
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 113..134
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 149..179
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 187..203
/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 211..227
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 497..909
/label=cTnT
/note="Chicken cardiac troponin T promoter"
intron 1036..1168
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
promoter 1213..1231
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1260..1976
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 2060..2648
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal 2739..2794
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3035..3051)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3193..3648
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3674..3778
/label=AmpR promoter
CDS 3779..4636
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4810..5398
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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