Adenovirus serotype 5 cloning vector. This plasmid contains a multiple cloning site in the E1 region to allow the insertion of transgene cassettes from any of our other plasmids.
Product Name: pSF-Ad5 (SnapFast Pro™)
Product Code: OG268
Size (bp): 33154 bp
Bacterial Antibiotic Selection:
Origin and Compatibility:
Bacterial Copy Number:
This is a serotype 5 adenovirus virus vector that allows the insertion of genes into the E1A position within the virus. The vector contains an E1 and E3 deletion and has been modified to remove a number of key restriction sites. The plasmid contains a small multiple cloning site consisting of AsiSI ClaI and PacI. These sites will be found in all of the plasmid vectors in our product range which allows the direct cloning of segments from our catalogue plasmids into the adenovirus genome (see cloning tab for more details).Packaging Capacity:
The wildtype virus genome is 35938 base pairs but in this vector the genome has been shortened by the deletions to make it 30501 base pairs. This allows for the insertion of up to 5.5kb of exogenous DNA. It has also previously been shown that serotype 5 adenovirus genomes can accomodate insertions up to 105% total genome length compared to wild type. For this reason it should be possible to insert an additional 1.7kb resulting in a total insertion size of 7.2Kb. We have previously packaged viruses up to 105% of the virus genome with good stability but some insertions may be unstable at this length depending on sequence. These numbers are therefore provided as guidelines only.Recovery and Growth:
Virus recovery requires two steps firstly the plasmid must be cleaved with the SwaI restriction enzyme to linearise the virus genome. Then the linear genome must be recovered and grown in 293 cells or any derivative of 293 cells such as 293As and 293T cells. These lines contain and express the adenovirus E1A protein that trans-compliments the deletion in the E1A region in this plasmid. A typically recovery transfection would use approximately 5ug of DNA. It is not necessary to remove the plasmid backbone after linearising the DNA a standard DNA clean up will be sufficient rather than gel extraction. Virus colonies should appear approximately 10-15 days after transfection into 293 cells however clones can sometimes appear at earlier time points. Cell should be cultured in 2% fetal calf serum to prevent over growth during this time.