pVITRO1-GFP/SEAP

• rEF1 and mEF1 prom: pVITRO1-GFP/LacZ plasmids carry two
elongation factor 1 alpha (EF-1α) promoters, from rat and mouse origins.
Both promoters display a strong activity that yields similar levels of expression. EF-1α promoters are expressed at high levels in all cell cycles and lower levels during G0 phase.
EF-1α promoters are also non-tissue specific; they are highly expressed in all cell types.
 

• SV40 enhancer, which is comprised of a 72-base-pair repeat, allows the enhancement of gene expression in a large host range. The enhancement varies from 2-fold in non-permissive cells to 20-fold in permissive cells. Furthermore, the SV40 enhancer is able to direct nuclear localization of plasmids.

• cMV enhancer: The major immediate early enhancer of the human cytomegalovirus (HCMV) is composed of unique and repeated sequence motifs. The HCMV enhancer can substitute for the 72-bp repeats of SV40 and is several-fold more active than the SV40 enhancer.

• SV40 pAn: The Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA.

• pMB1 Ori is a minimal E. coli origin of replication to limit vector size, but with the same activity as the longer Ori.

• FMDV IRES: The internal ribosome entry site of the Foot and Mouth Disease Virus enables the translation of two open reading frames from one mRNA with high levels of expression.
 

• EM7 is a bacterial promoter that enables the constitutive expression of the antibiotic resistance gene in E. coli.

• Antibiotic resistance gene : 
- Resistance to Blasticidin S is conferred by the bsr gene from Bacillus cereus. 
- The hph gene confers resistance to Hygromycin B both in E. coli and mammalian cells.
- The neo gene from Tn5 confers resistance to Kanamycin in E.coli and G418 in mammalian cells.
In bacteria, the resistance gene is expressed from the constitutive E. coli EM7 promoter. In mammalian cells, the resistance gene is transcribed from the rat EF-1α promoter as a polycistronic mRNA and translated via the FMDV IRES.

• EF1 pAn is a strong polyadenylation signal.

• GFP gene: This red-shifted variant of the jellyfish GFP gene encodes a green fluorescent protein that absorbs blue light (major peak at 480 nm) and emits green light (major peak at 505 nm).