Fly RMCE
P[attP.w+.attP] insertions
Below are descriptions of 4 insertions of P[attP.w+.attP], an RMCE target containing the mini-white gene flanked by inverted attP sites in which the UAS and TATA sequences of the original pUASTP2 target have been removed (note that this vector has also been referred to as pUASTP2.1). The insertion point is given relative to Release 4.3 of the genome sequence, and the distance to the nearest gene is provided. All 4 lines are homozygous viable and have been tested for RMCE. Other less-characterized insertions exist; please contact us if you are interested.
37B7
sequence from: 5' end
position: 18964658
strand: bottom
nearest gene: 8 bp upstream of Nak
38F1
sequence from: 3' end
position: 20810313
strand: top
nearest gene: 1.7 kb upstream of CG9331
53F8
sequence from: 3' end
position: 12612373
strand: top
nearest gene: 80 bp upstream of GstS1
89B8
sequence from: 3' end
position: 12051747
strand: bottom
nearest gene: 376 bp into 5' UTR of Tara
pUASTP2 insertions
Below are detailed descriptions of the 4 target insertions described in our manuscript, including the insertion point in release 4.3 of the genomic sequence, the position relative to the nearest gene, and a thumbnail of a 20 kb window centered on the point of insertion.
*Please note that the UAS binding sites and hsp70 promoter of pUAST are outside of the exchange cassette in these inserts - i.e. following RMCE, these elements will remain flanking the cassette. We are currently generating insertions of a new target site that lacks these elements.
25C
Sequence was generated by inverse pcr from the 3' end of the insert, and confirmed by sequencing PCR fragments across the 5' and 3' ends, which were generated using return primers in the flanking genomic sequence. The insertion point is on chromosome arm 2L at position 5006634 relative to the 5' end. The insert is within the Rtnl gene, 71bp upstream of the 5' end of the "B" transcript.
42A
Sequence was generated by inverse pcr from the 3' end of the insert, and confirmed by sequencing PCR fragments across the 5' and 3' ends, which were generated using return primers in the flanking genomic sequence;The insertion point is on chromosome arm 2R at position 1749343 relative to the 5' end. The insert is 69 bp upstream of Pld.
*Note that we see evidence of variegation with this insertion line; overall the mini-white expression is very weak, with patches of slightly stronger expression (very pale yellow with patches of stronger yellow; this pattern is more obvious as the flies age). The region of insertion is rich in natural transposons, particularly past the 3' end of the Pld gene.
52D
Sequence was generated by inverse PCR from the 3' end of the insert. The insertion point is on chromosome arm 2R at position 11507130. The insert maps 84 bp into the 5' end of predicted gene CG8315.
82F
Sequence was generated by inverse PCR from the 3' end of the insert. Insertion point is on chromosome arm 3R at position 1082377. The insert maps 4 bp upstream of predicted gene CG12173.
