From: Natalie Kuldell <nkuldell@MIT.EDU>
To: lrafty@dna20.com
Cc: nkuldell@mit.edu
Subject: new gene synthesis HEM1-HAm

Hi Louise-
I think I'm close to getting a new gene synthesis order ready. Can I ask for
your help in the planning. I know the ORF (S. cerevisiae HEM1) and tag (HA) I
want and I know that I want to express it in the yeast mitochondria. What I'm
working on still is the flanking sequence for integration.

It would be a great help in the planning if you could optimize the following
protein sequence for expression in S. cerevisiae mitochondria. Again, though
the sequence should have no EcoRI, XbaI, NotI, SpeI or PstI sites. I'll add
those in to the flanking region once I figure out how much of that I'll need.

Thanks as always for all help.
Natalie

PS Here's the protein sequence:

MQRSIFARFGNSSAAVSTLNRLSTTAAPHAKNGYATATGAGAAAATATASSTHAAAAAAA
AANHSTQESGFDYEGLIDSELQKKRLDKSYRYFNNINRLAKEFPLAHRQREADKVTVWCS
NDYLALSKHPEVLDAMHKTIDKYGCGAGGTRNIAGHNIPTLNLEAELATLHKKEGALVFS
SCYVANDAVLSLLGQKMKDLVIFSDELNHASMIVGIKHANVKKHIFKHNDLNELEQLLQS
YPKSVPKLIAFESVYSMAGSVADIEKICDLADKYGALTFLDEVHAVGLYGPHGAGVAEHC
DFESHRASGIATPKTNDKGGAKTVMDRVDMITGTLGKSFGSVGGYVAASRKLIDWFRSFA
PGFIFTTTLPPSVMAGATAAIRYQRCHIDLRTSQQKHTMYVKKAFHELGIPVIPNPSHIV
PVLIGNADLAKQASDILINKHQIYVQAINFPTVARGTERLRITPTPGHTNDLSDILINAV
DDVFNELQLPRVRDWESQGGLLGVGESGFVEESNLWTSSQLSLTNDDLNPNVRDPIVKQL
EVSSGIKQYPYDVPDYA stop