The pTagCFP-tubulin Expression Vector is a mammalian expression vector encoding the TagCFP-tubulin fusion protein. The vector can be used for fluorescent labeling of tubulin filaments in living cells.
TagCFP codon usage is optimized for high expression in mammalian cells, i.e. humanized. Human α-tubulin is fused to the TagCFP C-terminus. To increase TagCFP-tubulin mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of the TagCFP-tubulin coding sequence.
pTagCFP-tubulin is not intended as a cloning vector; however, the vector backbone contains unique restriction sites that permit excision of the TagCFP-tubulin hybrid sequence.
The vector backbone also contains immediate early promoter of cytomegalovirus (P
CMV IE) for protein expression, SV40 origin for replication in mammalian cells expressing SV40 T-antigen, pUC origin of replication for propagation in
E. coli, and f1 origin for single-stranded DNA production. SV40 polyadenylation signals (SV40 poly A) direct proper processing of the 3’ end of the reporter mRNA.
SV40 early promoter provides neomycin resistance gene expression to select stably transfected eukaryotic cells using G418. Bacterial promoter (P) provides kanamycin resistance gene expression in
E. coli. Kan
r/Neo
r gene is linked with herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signals.
| PRODUCT PROPERTIES |
| Application Notes: | | Expression in mammalian cells
pTagCFP-tubulin Expression Vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of the TagCFP-tubulin fusion in eukaryotic cells. If required, stable transformants can be selected using G418.
Propagation in E. coli
Suitable host strains for propagation in E. coli include DH5alpha, HB101, XL1-Blue, and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM109 or XL1-Blue. Plasmid incompatibility group is pMB1/ColE1. E. coli replication origin is pUC. The vector confers resistance to kanamycin (30 μg/ml) to E. coli hosts. Copy number in E. coli is about 500.
|
| Long Term Storage: | | -20°C |
| Background / Technical Information: | | For the Original Manufacturer's data sheet please click here. |
|
|
|
United States
