On Thursday, May 16, 2013 2:27:33 AM UTC-4, Avery wrote:
For those of you wondering "what the heck is in that plasmid anyways", I had the pGREEN plasmid from Carolina sequenced with M13 forward with m13 reverse.When BLAST-ed it comes up as: "Cloning vector pGFP, complete sequence"
here is the sequence:
GGGCGANTCGAGCTCGGTACCCGGGGNNCCGACGCGT
GGATCCTCAGTTGTACAGTTCATCCATGCCATGTGTAATCCCAGCAGCTG
TTACAAACTCAAGAAGGACCATGTGGTCTCTCTTTTCGTTGGGATCTTTC
GAAAGGGCAGATTGTGTGGACAGGTAATGGTTGTCTGGTAAAAGGACAGG
GCCATCGCCAATTGGAGTATTTTGTTGATAATGGTCTGCTAGTTGAACGC
TTCCATCTTCAATGTTGTGGCGGGTCTTGAAGTTCACTTTGATTCCATTC
TTTTGTTTGTCTGCCATGATGTATACATTGTGTGAGTTATAGTTGTATTC
CAATTTGTGTCCCAGAATGTTGCCATCTTCCTTGAAGTCAATACCTTTTA
ACTCGATTCTATTAACAAGGGTATCACCTTCAAACTTGACTTCAGCACGT
GTCTTGTAGTTGCCGTCATCTTTGAAGAAGATGGTCCTTTCCTGTACATA
ACCTTCGGGCATGGCACTCTTGAAAAAGTCATGCCGTTTCATATGATCCG
GGTATCTTGAAAAGCATTGAACACCATAGCACAGAGTAGTGACTAGTGTT
GGCCATGGAACAGGCAGTTTGCCAGTAGTGCAGATGAACTTCAGGGTAAG
TTTTCCGTATGTTGCATCACCTTCACCCTCTCCACTGACAGAGAACTTGT
GGCCGTTAACATCACCATCTAATTCAACAAGAATTGGGACAACTCCAGTG
AAGAGTTCTTCTCCTTTGCTAGCCATCTTTAGTCTAGAGTCGACCCTAGA
GCTTTTGTTCCCTTTAGTGAGGGTTAATTTCGAGCTTGGCGNAATCATGG
TCATAGCTGTTTCCTGNGTGAAATTGNTATNCGCTCACAATTNCANNCNA
CATNCGAGCCGGANNCATAAANTGNAAAGCCTGGGGNGCC Attached are the forward and reverse sequences for three samples. the ones that are 116060 should be forward reads and 116108 should be reverse.
Nice sequence. I also had sequenced pGreen a couple years ago. Our sequences agree except for a few mismatches at the 5 and 3' ends, probably due to bad sequence calls/quality.
Mine can be found here: http://www.roningenetics.org/Sequencing.html
Go to the bottom of the page under the heading "Sanger Sequencing". My consensus of the 4 chromatographs posted is the fasta file with the header "pGreen_consensus_seq" and at the very bottom there is an alignment of your sequence and mine. If you look through the patent for this (pdf on the website) the GFP insert is cloned into pBSSK, which is a blue/white vector, which should explain the lac sequence homology.
Tom
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