pHcRed1-1 Vector
pHcRed1-1 Vector
Brand: Takara Bio.
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pHcRed1-1 Vector
pHcRed1-1 is a promoterless, mammalian expression vector which can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into the multiple cloning site (MCS) located upstream of the HcRed1 coding sequence. Promoters should be cloned into the pHcRed1-1 MCS upstream from the HcRed1 coding sequences. Without the addition of a functional promoter, this vector will not express HcRed1.
HcRed1 is a far-red fluorescent protein which was generated by mutagenesis of a non-fluorescent chromoprotein from the reef coral Heteractis crispa. The HcRed1 coding sequence has been human codon-optimized for higher expression in mammalian cells. The sequence upstream of HcRed1 has been converted to a Kozak consensus sequence to further increase the translation efficiency in eukaryotic cells. SV40 polyadenylation signals downstream of the HcRed1 gene direct proper processing of the 3' end of the HcRed1 mRNA. The vector also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin resistance cassette (Neor)-consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene-allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette expresses kanamycin resistance in E. coli.
pHcRed1-1 can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418.
Overview
- Far-red fluorescent protein, ideal for in vivo imaging
- HcRed1 fluorescent protein excitation and emission maxima: 588 and 618 nm, respectively
- HcRed1 is detected on a Western blot using anti-RCFP polyclonal pan antibody
Applications
- Fusions
- Protein localization studies
- General reporter for mammalian cells
- Monitoring transfection efficiencies
- In vivo imaging
Vector map for promoterless fluorescent protein vectors AcGFP1-1, DsRed2-1, DsRed-Express-1, HcRed1-1 and ZsGreen1-1
Vector map for promoterless fluorescent protein vectors pAcGFP1-1, pDsRed2-1, pDsRed-Express-1, pHcRed1-1 and pZsGreen1-1.
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1.
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Vector map for promoterless fluorescent protein vectors AcGFP1-1, DsRed2-1, DsRed-Express-1, HcRed1-1 and ZsGreen1-1
Vector map for promoterless fluorescent protein vectors pAcGFP1-1, pDsRed2-1, pDsRed-Express-1, pHcRed1-1 and pZsGreen1-1.
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1.
