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pBABE-puro mCherry-EGFP-LC3B Notes

Addgene diagnostic digest

This gel depicts the NEB 1kb ladder and a digestion of the plasmid with the following enzymes (separate digests): NotI and SalI

We expected the following band sizes for this digest: 7KB (linearization) for both. We see the expected pattern, assuming the lower band in the SalI lane is undigested plasmid.

Sequence and user feedback from Dr. Ian Gentle,

"There is a significant difference in the psi pack sequence to what is generally given for pBABE
vectors on addgene.

In the PSI pack region that I sequenced, there is an 8Bp region not
present in the pbabe maps on the addgene website. This contains an extra
PstI site in it and seems to be present in many other PSI pack
sequences. I have attached...the sequence files for your info. At
the very least I thought this would be useful for others using this
plasmid, as it is impossible to pcr out the fusion protein using gene
specific primers as the mcherry and egfp are identical for the first
20bp, so an upstream sequence would be helpful for future purchasers."