FlyBase and the BDSC both currently have the sequence location of P{lacW}l(1)G0272G0272 (FBti0013149) as X:16 ,447,340..16,447,611 +. Ulrich Schaefer submitted a 400 bp flanking sequence that was determined using 3' inverse PCR. The first ~150 bp of this sequence have N's scattered through it. When I align the entire 400 bp to the release 6 reference sequence, I get a credible alignment of the segment from 20-400 to X:16447236-16447611 (+) (see below). Because this is a 3' flank, that would put the insertion site at X:16447216..16447236 , which is ~100 bp to the left of the range currently given by FlyBase and the BDSC. This places the insertion site ~ 223-243 bp upstream of the 5' end of the eIF2alpha gene. Plus Strand HSP: Score = 1345 (374 bits), P = 0 Identities = 340/381 (89.2%), Positives = 340/381 (89.2%), Strand = Plus / Plus Query: 20 ATGTTNATGTTTGTTAAANTTTATTGCGGTNAAANAAAANTTGCGATAATANCCCNATGA 79 ||||| || ||||||||| |||||||| | | |||| |||||| ||| || | | Sbjct: 16447236 ATGTTAATTTTTGTTAAA-TTTATTGCAATATTACAAAAATTGCGAAAATTGACCCAAAA 16447294 Query: 80 GTTNAATTCTCGAAAAGTTATATAGATTGCTAATACTGNATATATNAGGAATAAAAAGCA 139 ||| ||||||| |||||||||||||||||||||||||| ||| || |||||||||||||| Sbjct: 16447295 GTTAAATTCTCAAAAAGTTATATAGATTGCTAATACTG-ATAAAT-AGGAATAAAAAGCA 16447352 Query: 140 GTTGTTAAGTTATCAATATGTCACTTTTGAGCCAAGTTATGATGATAGTTCCCATAAGTC 199 |||||| ||||| ||||||||||||| | ||| |||||||||||| |||||||| ||| Sbjct: 16447353 GTTGTTCTTTTATCTATATGTCACTTTTTATCCATGTTATGATGATAATTCCCATATGTC 16447412 Query: 200 AAATGTTCATTTTTCATATTTATCTGTTTAAATTTGCAGTAACATTTTAAAAGGTTGCAT 259 |||| |||||||||||||||||||||||||||||||||| |||||||||||||||||||| Sbjct: 16447413 AAATTTTCATTTTTCATATTTATCTGTTTAAATTTGCAG-AACATTTTAAAAGGTTGCAT 16447471 Query: 260 ATAATTCGCAAACAAAAAGGCCATTAAATATGTAATGACATGTAGCTTAAATCGGTTTTT 319 || | ||||| ||||||||||||||||||||||||||||||||||||| ||||||||||| Sbjct: 16447472 ATTA-TCGCATACAAAAAGGCCATTAAATATGTAATGACATGTAGCTTGAATCGGTTTTT 16447530 Query: 320 TTAGAAGCTTCTCTTTTAATTTAAAGAACTTTAGAAAATTTGTATAGGTTGGCTCCGTTT 379 ||||||||| | ||||||||||||||||| |||||| ||||||||||||||||||||||| Sbjct: 16447531 TTAGAAGCTACACTTTTAATTTAAAGAACATTAGAATATTTGTATAGGTTGGCTCCGTTT 16447590 Query: 380 ATTAGTTAATTATGACAGCGC 400 ||||||||||||||||||||| Sbjct: 16447591 ATTAGTTAATTATGACAGCGC 16447611