Basic Vector Information
AAVS1 donor vector for genomic targeting
- Vector Name:
- AAVS1-Neo-M2rtTA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10080 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Promoter:
- CAG
AAVS1-Neo-M2rtTA 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 ℃.
AAVS1-Neo-M2rtTA vector Sequence
LOCUS 40924_170 10080 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10080)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 10080)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10080
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 33..836
/label=HA-L
/note="left homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
misc_feature 843..868
/label=SA
/note="splice acceptor site"
CDS 892..945
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 961..1761
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
polyA_signal 1836..2291
/label=PGK poly(A) signal
/note="mouse phosphoglycerate kinase 1 polyadenylation
signal"
enhancer 2389..2692
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2694..2971
/note="chicken beta-actin promoter"
intron 2973..3990
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
CDS 4090..4833
/label=rtTA-Advanced
/note="improved tetracycline-controlled transactivator"
polyA_signal complement(4846..4980)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 5310..6146
/label=HA-R
/note="right homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
promoter complement(6187..6205)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6212..6228)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS 6366..6665
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS complement(8068..8925)
/label=AmpR
/note="beta-lactamase"
rep_origin 9049..9637
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 9925..9946
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9961..9991
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9999..10015
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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 10023..10039
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 10060..10078
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
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