Re: [DIYbio] Re: Sequenced Carolina GFP plasmid

From: Avery louie <inactive.e@gmail.com>
To: "diybio@googlegroups.com" <diybio@googlegroups.com>
Cc: "diybio-eu@diybio.eu" <diybio-eu@diybio.eu>, "diybio-boston@googlegroups.com" <diybio-boston@googlegroups.com>
Sent: Sun, Sep 8, 2013 21:30:01 EDT
Subject: Re: [DIYbio] Re: Sequenced Carolina GFP plasmid

Which sequence is 900 bp?  It has been a while and I never really looked into it.  That said I definitely trust the sequence I have over Carolinas sequence.

--A

On Sun, Sep 8, 2013 at 12:56 AM, Maryam Saffarian <msaffar@g.clemson.edu> wrote:

Hi,

I was wondering that sequence is for gfP or for gfp-lacZ? According to pGreen Carolina lab, mutant gfp fused to lacZ is around 600bp but the sequence above is 900bp. I am trying to design primers for the mutant gfp fused to lacZ.

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.

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