| pcDNA3.1/V5-His TOPO |
Mammalian expression vector |
Invitrogen, Carlsbad, CA |
| pcDNA3 |
Mammalian expression vector vector |
Invitrogen, Carlsbad, CA |
| pcDNA3-VEGF121
|
pcDNA3 encoding human VEGF121
|
Weber, unpublished |
| pCF10 |
Dual-regulated expression vector |
Fux and Fussenegger, submitted for publication |
| pEF4/MycHisB |
Expression vector containing the human elongation factor 1α promoter (PEF1α ) |
Invitrogen, Carlsbad, CA |
| pEYFP-C1 |
Expression vector encoding the enhanced yellow fluorescent protein under control of PhCMV
|
Clontech, Palo Alto, CA |
| PMSCVneo |
Retroviral expression vector derived from the murine embryonic stem cell virus |
Grez et al., 1990 |
| PLPCX |
Retroviral expression vector derived from the Moloney MLV (MoMuLV) and Moloney murine sarcoma virus (MoMuSV) |
Miller and Rosman, 1989 |
| pNL-EGFP |
Lentiviral expression vector (5′LTR-ψ+-oriSV40-cPPT-RRE-3′LTR) |
Mochizuki et al., 1998 |
| pNL-EGFPΔU3 |
pNL-EGFP containing an EcoRV/PvuII deletion in the U3 region of the 3′LTR eliminating any enhancers (5′LTR-ψ+-oriSV40-cPPT-RRE-3′LTRΔU3) |
Reiser et al., unpublished |
| pSS173 |
Mammalian expression vector encoding the human placental SEAP |
Schlatter et al., 2002 |
| pWW85 |
Triple-fluorescent expression vector encoding ECFP, RFP and EYFP |
Weber et al., 2002 |
| pWW265 |
VEGF121 was excised from pcDNA-VEGF121 by HindIII/SpeI and ligated into the corresponding sites (HindIII/SpeI) of pCF10 (PhCMV-VEGF121-pA) |
Weber et al., unpublished |
| pTRIDENT1 |
Tricistronic mammalian expression vector harboring a tetracycline-responsive, PhCMV*-1-driven expression unit (PhCMV*-1-MCSI-IRESI-MCSII-IRESII-MCSIII-pA) (pMF125) |
Fussenegger et al., 1998 |
| pTRIDENT3 |
Tricistronic mammalian expression vector harboring a tetracycline-responsive, PhCMV*-1-driven expression unit (PhCMV*-1-MCSI-IRES-MCSII-CITE*-MCSIII-pA) (pMF122) |
Fussenegger et al., 1998 |
| pMF201 |
Dual-regulated expression vector encoding EYFP and ECFP |
Fussenegger et al., 2000 |
| pMF242 |
pcDNA3.1/V5-His TOPO encoding mouse erythropoietin (mEPO) |
Fussenegger et al., 2000 |
| pMF320 |
PEF1α was amplified from pEF4/MycHisB by OMF164/OMF165 and cloned in antisense orientation into pcDNA.3.1/V5-His TOPO |
This work |
| pMF351 |
PhCMV-EYFP was amplified from pEYFP-C1 with OMF179/OMF180 and cloned as HpaI/KpnI fragment into the corresponding sites (HpaI/KpnI) of pNL-EGFPΔU3 (5′LTR-ψ+-oriSV40-cPPT-RRE-MCSI-PhCMV-EYFP-MCSII-3′LTRΔU3) |
This work |
| pMF352 |
U3 was amplified from pNL-EGFPΔU3 using OMF183/OMF184 and cloned into pcDNA3.1/V5-His TOPO. The U3 fragment contains a SbfI site |
This work |
| pMF353 |
R-U5-ψ+-SD was amplified from pNL-EGFPΔU3 using OMF185/OMF186 and cloned into pcDNA3.1/V5-His TOPO |
This work |
| pMF355 |
PPT-3′LTRΔU3 was amplified from pNL-EGFPΔU3 using OMF189/OMF190 and cloned into pcDNA3.1/V5-His TOPO |
This work |
| pMF356 |
The PhCMV-EYFP cassette of pMF351 was excised by NotI and the backbone religated (5′LTR-ψ+-oriSV40-cPPT-RRE-MCS-3′LTRΔU3) |
This work |
| pMF357 |
U3 was excised from pMF352 by XhoI/SrfI and cloned into the corresponding sites (XhoI/SrfI) of pTRIDENT1 |
This work |
| pMF358 |
R-U5-ψ+-SD was excised from pMF353 by SrfI/SwaI and cloned into the corresponding sites (SrfI/SwaI) of pMF357 |
This work |
| pMF358x |
R-U5-ψ+-SD was excised from pMF376 by SrfI/SwaI and cloned into the corresponding sites (SrfI/SwaI) of pMF357 |
This work |
| pMF359 |
PEF1α was excised from pMF320 by EcoRV/XmaI and cloned into the corresponding sites (HpaI/XmaI) of pMF356 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-MCS-3′LTRΔU3) |
This work |
| pMF360 |
cPPT-RRE-SA was excised from pMF375 by SwaI/MluI and cloned into the corresponding sites (SwaI/MluI) of pMF358 |
This work |
| pMF360x |
cPPT-RRE-SA was excised from pMF375 by SwaI/MluI and cloned into the corresponding sites (SwaI/MluI) of pMF358x |
This work |
| pMF363 |
PhCMV was excised from pLPCX using StuI/EcoRI and cloned into the compatible sites (HpaI/EcoRI of pMF356 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-MCS-3′LTRΔU3) |
This work |
| pMF365 |
EYFP was excised from pMF201 by EcoRV/SpeI and ligated to the compatible sites (SmaI/SpeI) of pMF359 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-EYFP-3′LTRΔU3) |
This work |
| pMF371 |
ECFP was excised from pMF201 by EcoRI/NotI and ligated to the corresponding sites (EcoRI/NotI) of pMF363 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-ECFP-3′LTRΔU3) |
This work |
| pMF375 |
cPPT-RRE-SA was amplified from pNL-EGFPΔU3 using OMF191/OMF192 and cloned into pcDNA3.1/V5-His TOPO |
This work |
| pMF376 |
R-U5-ψ+-SD was amplified from pNL-EGFPΔU3 using OMF185/OMF193 and cloned into pcDNA3.1/V5-His TOPO |
This work |
| pMF378 |
OriSV40 was amplified from pNL-EGFPΔU3 using OMF195/OMF196 and cloned into pcDNA3.1/V5-His TOPO |
This work |
| pMF388 |
The 5′LTR-ψ+-oriSV40-cPPT-RRE-MCS-3′LTRΔU3 was amplified from pMF356 by OMF199/OMF200 and cloned as AatII/NheI fragment to the compatible sites (AatII/XbaI) of the pTRIDENT1 backbone (5′LTR-ψ+-oriSV40-cPPT-RRE-MCS-3′LTRΔU3) |
This work |
| pMF394 |
PhCMV-EYFP was excised from pMF351 using HpaI/KpnI and cloned into the corresponding sites (HpaI/KpnI) of pMF388 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-EYFP-3′LTRΔU3) |
This work |
| pMF402 |
PPT-3′LTRΔU3 was excised from pMF355 by MluI/XbaI and cloned into the corresponding sites (MluI/XbaI) of pMF360 |
This work |
| pBM1 |
The ECFP-IRESI-RFP cassette was excised from pWW85 by EcoRI/AscI and cloned into the corresponding sites of pLentiTRIDENT2 (EcoRI/AscI) 5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-ECFP-IRESI-RFP-IRESII-MCSIII-3′LTRΔU3
|
This work |
| pBM6 |
The PPGK-neo cassette was amplified from pMSCVneo using OBM1/OBM2 and cloned in sense orientation into pcDNA3.1/V5-His TOPO |
This work |
| pMB7 |
PEF1α was excised from pMF320 using EcoRV/XmaI and cloned into the compatible sites (HpaI/XmaI) of pMF388 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-MCS-3′LTRΔU3) |
This work |
| pBM8 |
The PPGK-neo cassette was excised from pBM6 by PacI/MluI and cloned into the corresponding sites (PacI/MluI) of pMF356 (5′LTR-ψ+-oriSV40-cPPT-RRE-MCSI-PPGK-neo-MCSII-3′LTRΔU3) |
This work |
| pBM9 |
The PPGK-neo cassette was excised from pBM6 by PacI/MluI and inserted into the corresponding sites (PacI/MluI) of pMF359 (5′LTR-ψ+-oriSV40-cPPT-RRE-PEF1α-MCSI-PPGK-neo-MCSII-3′LTRΔU3) |
This work |
| pBM13 |
The PPGK-neo cassette was excised from pBM6 using PacI/MluI and cloned into the corresponding sites (PacI/MluI) of pMF388 (5′LTR-ψ+-oriSV40-cPPT-RRE-MCSI-PPGK-neo-MCSII-3′LTRΔU3) |
This work |
| pBM14 |
The PPGK-neo cassette was excised from pBM6 using PacI/MluI and cloned into the corresponding sites (PacI/MluI) of pBM7 (5′LTR-ψ+-oriSV40-cPPT-RRE-PEF1α-MCSI-PPGK-neo-MCSII-3′LTRΔU3) |
This work |
| pBM40 |
EYFP was excised from pLentiModule4 by NheI/MluI and cloned into the compatible sites (SpeI/MluI) of pBM7 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-EYFP-3′LTRΔU3) |
This work |
| pBM42 |
mEPO was excised from pMF242 by EcoRI/EcoRV and cloned into the compatible sites (EcoRI/PmeI) of pMF359 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-EPO-3′LTRΔU3) |
This work |
| pBM43 |
VEGF121 was excised from pWW265 by SalI/MluI and ligated into the corresponding sites (SalI/MluI) of pMF359 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-VEGF121-3′LTRΔU3) |
This work |
| pBM44 |
SEAP was excised from pSS173 by EcoRI/EcoRV and ligated into the compatible sites (EcoRI/PmeI) of pMF359 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhEF1α-SEAP-3′LTRΔU3) |
This work |
| pLentiTRIDENT1 |
The MCSI-IRESI-MCSII-IRESII-MCSIII cassette was excised from pTRIDENT1 by EcoRI/BglII and cloned into the compatible sites (EcoRI/BamHI) of pMF363 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-MCSI-IRESI-MCSII-IRESII-MCSIII-3′LTRΔU3) (pMF370) |
This work |
| pLentiTRIDENT2 |
The MCSI-IRES-MCSII-CITE*-MCSII cassette was excised from pTRIDENT3 by EcoRI/BglII and cloned into the compatible sites (EcoRI/BamHI) of pMF363 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-MCSI-IRES-MCSII-CITE*-MCSIII-3′LTRΔU3) (pMF369) |
This work |
| pLentiTFT1 |
The triple-fluorescent expression cassette ECFP-IRESI-RFP-IRESII-EYFP was excised from pWW85 using EcoRI/MluI and cloned into the corresponding sites (EcoRI/MluI) of pLentiTRIDENT1 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-ECFP-IRESI-RFP-IRESII-EYFP-3′LTRΔU3) (pMB2) |
This work |
| pLentiTFT2 |
EYFP was excised from pWW85 by SpeI/MluI and cloned to the corresponding sites (SpeI/MluI) of pBM1 (5′LTR-ψ+-oriSV40-cPPT-RRE-PhCMV-ECFP-IRES-RFP-CITE*-EYFP-3′LTRΔU3) (pMB3) |
This work |
| pLentiModule1 |
PPT-3′LTRΔU3 was excised from pMF355 by MluI/XbaI and cloned into the corresponding sites (MluI/XbaI) of pMF360x (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-MCS-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI) (pBM4) |
This work |
| pLentiModule2 |
The PhCMV-EYFP cassette was excised from pMF351 by EcoRI/MluI and cloned into the corresponding sites (EcoRI/MluI) of pLentiModule1 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-PhCMV-EYFP-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI) (pBM5) |
This work |
| pLentiModule3 |
oriSV40 was excised from pMF378 by XbaI/NheI and cloned into the XbaI sites of pLentiModule1 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-MCS-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI-oriSV40) (pBM15) |
This work |
| pLentiModule4 |
oriSV40 was excised from pMF378 by XbaI/NheI and cloned into the XbaI sites of pLentiModule2 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-PhCMV-EYFP-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI-oriSV40) (pBM12) |
This work |
| pLentiModule5 |
The PhEF1α-EYFP cassette was excised from pMF365 by BamHI and ligated in sense orientation into the corresponding site (BamHI) of pLentiModule1 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-PhEF1α-EYFP-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI) (pBM45) |
This work |
| pLentiModule6 |
The PhEF1α-VEGF121 cassette was excised from pBM43 by BamHI and ligated in sense oritentation into the corresponding site (BamHI) of pLentiModule1 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-PhEF1α-VEGF121-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI) (pBM46) |
This work |
| pLentiModule7 |
The PhEF1α-EPO cassette was excised from pBM42 by BamHI and ligated in sense orientation into the corresponding site (BamHI) of pLentiModule1 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-PhEF1α-EPO-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI) (pBM47) |
This work |
| pLentiModule8 |
The PhEF1α-SEAP cassette was excised from pBM44 by BamHI and ligated in sense orientation into the corresponding site (BamHI) of pLentiModule1 (XhoI/NruI-U31-StuI/SbfI-U32-SrfI-R-U5-ψ+-SD-SwaI-cPPT-RRE-SA-PhEF1α-SEAP-ΔU31-PmeI/HpaI/BclI-ΔU32-NdeI/NheI/SalI/XbaI) (pBM48) |
This work |
