Subject: ebi constructs and insertions
The following information was provided by Annette Parks of Exelixis, Inc.
In P{UAS-ebi.Exel}, wild type ebi (FBgn0023444) coding sequences were
cloned into P{Express-UAS}.
P{UAS-ebi.Exel}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-ebi.Exel}2 is a second chromosome insertion.
P{UAS-ebi.Exel}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-ebi.HA.Sma}, ebi coding sequence with HA tag sequence placed into
a unique SmaI site was cloned into P{Express-UAS}. The protein variant has
the HA tag (YPYDVPDYASL) approximately 100 amino acids from the N terminus
to give the internal sequence VKTEPGK-HAtag-P.
P{UAS-ebi.HA.Sma}2 is a homozygous viable and fertile, second chromosome
insertion.
P{UAS-ebi.HA.Sma}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-ebi.HA.Nco}, ebi coding sequence with HA tag sequence placed into
a unique NcoI site was cloned into P{Express-UAS}. The protein variant has
the HA tag (YPYDVPDYASL) approximately 17 amino acids N terminal of the WD1
repeat to give the internal sequence VPM-HAtag-M (the M is duplicated).
P{UAS-ebi.HA.Nco}2 is a homozygous viable and fertile, second chromosome
insertion.
P{UAS-ebi.HA.Nco}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-ebi.DN}, a truncated ebi sequence was cloned into
P{Express-UAS}. The construct encodes a dominant negative protein variant
that is essentially only the WD repeats at the C terminus. The ebi
sequence starts GCCGCGCCCCGTCATGACTCCCGCAGCTCTTGTTCCCATGGACA. The initial
GCCGCGCC is an artificial AcsI site. This is followed by a start
ATG. This is then followed by the truncated ebi.
P{UAS-ebi.DN}3 is a homozygous viable and fertile, third chromosome insertion.
In P{ebi.FRT}, ebi coding sequences and upstream regulatory regions were
cloned into P{Express-FRT.2.1}. Upon exposure to FLP recombinase,
miniwhite and the ebi gene are excised from the construct.
P{ebi.FRT}1 is a homozygous and hemizygous viable and fertile, X chromosome
insertion.
P{ebi.FRT}2 is a homozygous viable and fertile, second chromosome insertion.
P{ebi.FRT}3 is a third chromosome insertion.