12275 bp - lentiviral vectorscfm.brc.riken.jp/wp-content/uploads/subteam_for...cs-rfa-etv 12275 bp...
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CS-RfA-ETV12275 bp
MluI 228
NotI 1521
NotI 2698
EcoRI 3019
MluI 3466
PstI 4141
AgeI 4375
ApaI 4894AgeI 5508
XbaI 5524ApaI 6208PstI 6418
XhoI 6946
ApaI 8257
CMV RU55' SD
RRE3' SA
cPPT+CTS
RfA
attR1
Cm
ccdBattR2
EF-1
TetRT2A
VenusPRE
del U3R
U5BGH pA
f1 ori
SV40 pro/ori
Zeo
SV40 pA
ColE1 ori
Amp 1
Miyoshi
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Plasmid name: CS-RfA-ETV Plasmid size: 12275 bp Note CMV: Human cytomegalovirus immediate early promoter y: Packaging signal 5’SD: 5’ splicing donor site 3’SA: 3’ splicing acceptor site RRE: Rev responsive element cPPT: Central polypurine tract CTS: Central termination sequence RfA: Gateway Reading Frame Cassette A containing Cm, ccdB, attR1, and attR2 Cm: Chloramphenicol resistance gene ccdB: The ccdB gene encodes a protein that interferes with E. coli DNA gyrase attR1, 2: Sites for site-specific recombination with a Gateway vector (Invitrogen). attR sites always recombine with attL sites in a reaction mediated by the LR Clonase (Invitrogen). EF-1: Human elongation factor 1 α subunit promoter TetR: Tet repressor T2A: Thosea asigna virus 'self-cleaving' 2A peptide sequence Venus: A variant of yellow fluorescent protein (YFP) gene PRE: Woodchuck hepatitis virus posttranscriptional regulatory element del U3: Deletion of enhancer and promoter sequences in the U3 region BGH pA: Bovine growth hormone polyadenylation signal SV40 pro & ori: SV40 early promoter and origin Zeo: Zeocin resistance gene SV40 pA: SV40 polyadenylation signal Amp: Ampicillin resistance gene *Use the DB3.1 or ccdB Survival 2 T1R E. coli strain for propagation of plasmids containing the ccdB gene.
Miyoshi
テキストボックス
y