Supplementary Figure 1: Sequencing of targeted alleles
(A) Overview of the APD, APD-PGK and APD-TET alleles. Sequences of mouse
origin are shown in black, sequences of vector origin in blue, the PGK promoter in
orange, the TET response element (TRE) in green and the CMV2 promoter in dark
green. Exon 3 of Igf2r is shown as a black bar and the loxP511 site left behind after
cassette removal is shown as a blue bar. The main transcriptional start site of Airn
(T1) is shown as black arrow. The PCR primers used to amplify the region for
sequencing (F1, R2) are shown as small black arrows.
(B) Sequences of the APD, APD-PGK and APD-TET allele. Sequences of mouse
origin are shown as black or white text, sequences of vector origin as blue text. The
PGK promoter is shown as orange underlined orange text. The TRE is shown as light
green underlined light green text, the CMV2 promoter as dark green underlined dark
green text. LoxP511 sites are shown as blue underlined bold blue text. Exon 3 of Igf2r
is shown as black box. The main transcriptional start site of Airn (T1) is shown as a
black arrow. Primers used to amplify the region for sequencing (F1, R2) are shown as
black or white bold text underlined with a black arrow. The APD allele contains
131bp of vector sequence including one loxP511 site 5’ of T1. The APD-PGK allele
contains 182bp of vector sequence including one loxP511 site followed by 506bp of
PGK promoter sequence. BLAT analysis of the PGK promoter sequence using the
UCSC genome browser matches the PGK promoter sequence on mouse chromosome
X, bp 103382068-103382574 (mm9). The PGK promoter sequence is followed again
by 37bp of vector origin. The APD-TET allele contains 124bp of vector sequence
including one loxP511 site followed by 317bp of TRE and 119bp of the CMV2
promoter (both from pTETsplice, Invitrogen) followed by 3bp of vector sequence.