pHcRed1-1 Vector

Description

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 coralHeteractis 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 inE. 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 inE. coli.pHcRed1-1 can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1. Back Vector map for promoterless fluorescent protein vectors pAcGFP1-1, pDsRed2-1, pDsRed-Express-1, pHcRed1-1 and pZsGreen1-1.