CSIV-TRE-RfA-CMV-KT Plasmid size
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CSIV-TRE-RfA-CMV-KT 11988 bp MluI 228 NotI 1521 EcoRI 2565 NheI 2905 NotI 3039 BamHI 3110 BspEI 3356 EcoRI 3360 MluI 3807 BamHI 3813 PstI 4482 AgeI 4625 XhoI 4631 EcoRI 5396 AgeI 5402 XbaI 6124 ApaI 6266 BamHI 6876 XbaI 6882 HpaI 6888 EcoRV 7736 BspEI 7776 ApaI 7970 PmeI 7976 CMV R U5 5' SD RRE 3' SA cPPT+CTS TRE P RfA attR1 Cm ccdB attR2 CMV hKO1 T2A rtTA PRE del U3 loxP R U5 BGH pA f1 ori SV40 pro & ori Zeo SV40 pA ColE1 ori Amp 1
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CSIV-TRE-RfA-CMV-KT11988 bp
MluI 228
NotI 1521
EcoRI 2565
NheI 2905NotI 3039BamHI 3110BspEI 3356EcoRI 3360
MluI 3807BamHI 3813
PstI 4482AgeI 4625
XhoI 4631EcoRI 5396
AgeI 5402XbaI 6124ApaI 6266
BamHI 6876XbaI 6882HpaI 6888
EcoRV 7736BspEI 7776
ApaI 7970PmeI 7976
CMV RU55' SD
RRE
3' SAcPPT+CTS
TRE P
RfA
attR1
Cm
ccdBattR2
CMVhKO1T2A
rtTAPRE
del U3loxPR
U5BGH pA
f1 ori
SV40 pro & ori
Zeo
SV40 pA
ColE1 ori
Amp 1
Miyoshi
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Plasmid name: CSIV-TRE-RfA-CMV-KT Plasmid size: 11988 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 TRE P: Tet-responsive promoter (TRE-Tight) 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). hKO1: Humanized Kusabira-Orange T2A: Thosea asigna virus 2A peptide sequence rtTA: Reverse Tet-controlled transactivator protein (Tet-On-Advanced) PRE: Woodchuck hepatitis virus posttranscriptional regulatory element del U3: Deletion of enhancer and promoter sequences in the U3 region loxP: Site-specific recombination site for Cre recombinase 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