LOCUS Ple104\(pEMS1084) 19808 bp DNA circular 31-JAN-2011 SOURCE ORGANISM COMMENT NOTE: Insert was not sequence confirmed, ends may be different and may contain some internal SNPs (sequence is based on initial cloning design, and not necessarirly final cloning design) COMMENT This file is created by Vector NTI http://www.invitrogen.com/ COMMENT ORIGDB|GenBank COMMENT VNTDATE|580573252| COMMENT VNTDBDATE|580573252| COMMENT LSOWNER| COMMENT VNTNAME|Ple104 (pEMS1084)| COMMENT VNTAUTHORNAME|Russell Bonaguro| COMMENT Vector_NTI_Display_Data_(Do_Not_Edit!) COMMENT (SXF COMMENT (CGexDoc "Ple104 (pEMS1084)" 0 19808 COMMENT (CDBMol 0 0 1 1 1 0 0 1 0 "" "" 0 0 0 0 (CObList) (CObList) (CObList) COMMENT (CObList) -1 "") COMMENT (CDocSetData 1 1 0 1 0 0 "MAIN" 1 1 1 1 0 0 1 1 0 1 10 10 4294967295 50 0 COMMENT 1 0 (CHomObj 0 0 0 3 75) (CWordArray) (CWordArray) (CStringList) COMMENT (CStringList "atg" "gtg") (CStringList "taa" "tga" "tag") (CObList) 1 COMMENT "{(0,1),2}" 0 0 "" 0 4294967295 0 0 0 0 0 0 0 "MAIN" 0 0 30 0 COMMENT (CProteinMotifSearchObject 70 20 1 1 1 1 0 0 1 0 0 0 0 0)) COMMENT (CMolPar 0 0 0 0 4294967295 1 19808 0 0 0 0 0 0 0 0) (CStringList) COMMENT (CStringList) (CObList) (COAPar 25 250 50 0 20 20 3 7) COMMENT (COAPar 25 250 50 0 6 4 3 7) (COAPar 25 250 50 0 6 4 3 7) (CObList) COMMENT (CObList COMMENT #0=(CFSignal (CObList) "pBS246 vector" 21 0 0 17494 137 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "17494..137") COMMENT #1=(CFSignal (CObList) "5' junction" 86 0 0 13670 17481 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "13670..17481") COMMENT #2=(CFSignal (CObList) "AmpR" 22 0 0 18801 19801 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "18801..19801") COMMENT #3=(CFSignal (CObList) "AmpR" 22 0 0 12198 13644 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "12198..13644") COMMENT #4=(CFSignal (CObList) "HPRT 3' homology" 86 0 0 9312 12202 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "9312..12202") COMMENT #5=(CFSignal (CObList) "HPRT complementary sequence" 86 0 0 4605 9311 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "4605..9311") COMMENT #6=(CFSignal (CObList) "FRT (F5 mutant)" 21 0 0 2497 2530 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "2497..2530") COMMENT #7=(CFSignal (CObList) "Kozak consensus" 21 0 0 2531 2542 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "2531..2542") COMMENT #8=(CFSignal (CObList) "eGFP (+mutated stop)" 21 0 0 2543 3259 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "2543..3259") COMMENT #9=(CFSignal (CObList) "Cre" 21 0 0 3260 4285 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "3260..4285") COMMENT #10=(CFSignal (CObList) "NLS" 21 0 0 4286 4358 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "4286..4358") COMMENT #11=(CFSignal (CObList) "FRT" 21 0 0 4362 4395 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "4362..4395") COMMENT #12=(CFSignal (CObList) "SV40 early polyA signal" 21 0 0 4549 4599 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "4549..4599") COMMENT #13=(CFSignal (CObList) "Prom region" 21 0 0 1612 2488 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "1612..2488") COMMENT #14=(CFSignal (CObList) "oEMS2380" 27 0 0 1874 1895 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "1874..1895") COMMENT #15=(CFSignal (CObList) "oEMS2400" 27 0 0 1345 1364 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "1345..1364") COMMENT #16=(CFSignal (CObList) "Ple104 (HAP1-B)" 21 0 0 160 2488 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "160..2488")) (CObList) COMMENT (CObList COMMENT #17=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "gcttcttctgtatgccctgttctg" 63 1245 24 0 100 100 100 COMMENT 56.1292 50 -178.1 -460.9 -38.9027 -11.9661) COMMENT (CPrimer 0 "ccactcgacgtcttccatttga" 63 1874 22 1 100 100 100 COMMENT 56.4216 50 -162.2 -415.2 -36.6281 -13.9149) 171 1245 1895 651 COMMENT 81.7278 55.7604 59.1482) COMMENT #18=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "cttcttctgtatgccctgttctgtg" 63 1246 25 0 100 100 100 COMMENT 56.0743 48 -179.3 -464.4 -39.0592 -11.7096) COMMENT (CPrimer 0 "ccactcgacgtcttccatttga" 63 1874 22 1 100 100 100 COMMENT 56.4216 50 -162.2 -415.2 -36.6281 -13.9149) 171 1246 1895 650 COMMENT 81.6983 55.6923 59.1111) COMMENT #19=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "tcttctgtatgccctgttctgtgc" 63 1248 24 0 100 100 100 COMMENT 57.1897 50 -173.5 -446.3 -38.6557 -13.507) COMMENT (CPrimer 0 "ccactcgacgtcttccatttga" 63 1874 22 1 100 100 100 COMMENT 56.4216 50 -162.2 -415.2 -36.6281 -13.9149) 171 1248 1895 648 COMMENT 81.7023 55.7099 59.2181) COMMENT #20=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "cttctgtatgccctgttctgtgct" 63 1249 24 0 100 100 100 COMMENT 56.4518 50 -175.7 -453.6 -38.6792 -13.1611) COMMENT (CPrimer 0 "ccactcgacgtcttccatttga" 63 1874 22 1 100 100 100 COMMENT 56.4216 50 -162.2 -415.2 -36.6281 -13.9149) 171 1249 1895 647 COMMENT 81.736 55.796 59.2417)) (CObList) (CObList) (CObList) (CObList) COMMENT (CTextView 0 COMMENT #21=(CGroupPar (CParagraph 0 (0 0) 1 2 0 0 180) COMMENT (CObjectList COMMENT #22=(CRefLinePar COMMENT (CLinePar (CParagraph 0 (0 0) 0 2 0 1 233) COMMENT "Ple104 (pEMS1084)" 2) 5 "" 0 4) COMMENT #23=(CFolderPar COMMENT (CGroupPar (CParagraph 1 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #24=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "DNA 'Ple104 (pEMS1084)'" 1) COMMENT #25=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Local object" 1) COMMENT #26=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Created: 01/31/11 02:20PM" 1) COMMENT #27=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Last Modified: 01/31/11 02:20PM" 1) COMMENT #28=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "length: 19808 bp" 1) COMMENT #29=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "storage type: Basic" 1) COMMENT #30=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "form: Circular" 1))) "General Description") COMMENT #31=(CFolderPar COMMENT (CGroupPar (CParagraph 2 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #32=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Original Source Database: GenBank" 1) COMMENT #33=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Modification Date in the Original DB: 13-MAR-2007" 1))) COMMENT "Standard Fields") COMMENT #34=(CRefLinePar COMMENT (CLinePar (CParagraph 0 (0 0) 0 2 0 0 233) "Comments" 2) 1 "" COMMENT 0 0) COMMENT #35=(CFolderPar COMMENT (CGroupPar (CParagraph 8 (0 0) 1 2 0 0 178) (CObjectList)) COMMENT "Annotations") COMMENT #36=(CFolderPar COMMENT (CGroupPar (CParagraph 12 (6 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #37=(CFolderPar COMMENT 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180) COMMENT " 4286..4358" 1))) "NLS") COMMENT #66=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 4362 (3 #11# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #67=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 4362 End: 4395" 1) COMMENT #68=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #69=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 4362..4395" 1))) "FRT") COMMENT #70=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 4549 (3 #12# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #71=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 4549 End: 4599" 1) COMMENT #72=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #73=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 4549..4599" 1))) COMMENT "SV40 early polyA signal") COMMENT #74=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 17494 (3 #0# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #75=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 17494 End: 137 " 1) COMMENT #76=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #77=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 17494..137" 1))) "pBS246 vector"))) COMMENT "Misc. Feature (10 total)") COMMENT #78=(CFolderPar COMMENT (CGroupPar (CParagraph 22 (7 22 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #79=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 12198 (3 #3# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #80=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 12198 End: 13644" 1) COMMENT #81=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #82=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 12198..13644" 1))) "AmpR") COMMENT #83=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 18801 (3 #2# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #84=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 18801 End: 19801" 1) COMMENT #85=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #86=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 18801..19801" 1))) "AmpR"))) COMMENT "Misc. Marker (2 total)") COMMENT #87=(CFolderPar COMMENT (CGroupPar (CParagraph 27 (7 27 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #88=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1345 (3 #15# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #89=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1345 End: 1364" 1) COMMENT #90=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) "56.3" COMMENT 1) COMMENT #91=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #92=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1345..1364" 1))) "oEMS2400") COMMENT #93=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1874 (3 #14# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #94=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1874 End: 1895" 1) COMMENT #95=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #96=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1874..1895" 1))) "oEMS2380"))) COMMENT "Primer (2 total)") COMMENT #97=(CFolderPar COMMENT (CGroupPar (CParagraph 86 (7 86 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #98=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 4605 (3 #5# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #99=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 4605 End: 9311" 1) COMMENT #100=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Consists of human HPRT promoter, human Exon1, Intron1, Exon2, and part of Intron2" COMMENT 1) COMMENT #101=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #102=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 4605..9311" 1))) COMMENT "HPRT complementary sequence") COMMENT #103=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 9312 (3 #4# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #104=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 9312 End: 12202" 1) COMMENT #105=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Consists of part of Intron 2, all of Exon 3, and part of Intron 3" COMMENT 1) COMMENT #106=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #107=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 9312..12202" 1))) COMMENT "HPRT 3' homology") COMMENT #108=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 13670 (3 #1# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #109=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 13670 End: 17481" 1) COMMENT #110=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "The 5 HPRT probe anneals to genomic sequence ~560bp upstream of the BamHI site at 9263" COMMENT 1) COMMENT #111=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #112=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 13670..17481" 1))) "5' junction"))) COMMENT "Misc. 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COMMENT " Tm: 81.7 C TaOpt: 59.1 C GC: 55.8" 1) COMMENT #121=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #122=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #123=(COligo "" COMMENT "GCTTCTTCTGTATGCCCTGTTCTG" COMMENT "Sense primer for PCR amplification of a region of Ple104 (pEMS1084)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " GCTTCTTCTGTATGCCCTGTTCTG" 1)) COMMENT #124=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #125=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 24 Tm: 56.1 C GC: 50.0" 1) COMMENT #126=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -178.1 kcal/mol dS: -460.9 cal/mol dG: -38.9 kcal/mol" COMMENT 1) COMMENT #127=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #128=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #129=(COligo "" "CCACTCGACGTCTTCCATTTGA" COMMENT "Antisense primer for 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Primer: " 1) COMMENT #146=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #147=(COligo "" "CCACTCGACGTCTTCCATTTGA" COMMENT "Antisense primer for PCR amplification of a region of Ple104 (pEMS1084)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " CCACTCGACGTCTTCCATTTGA" 1)) COMMENT #148=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #149=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.4 C GC: 50.0" 1) COMMENT #150=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -162.2 kcal/mol dS: -415.2 cal/mol dG: -36.6 kcal/mol" COMMENT 1) COMMENT #151=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 0.3" 1) COMMENT #152=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 2.0" 1))) COMMENT "#2: Product of length 650 (rating: 171)")) COMMENT #153=(CPCRFolderPar COMMENT (CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 0 COMMENT (20 COMMENT #154=(CPCRData (CStringList) (CStringList) COMMENT "tcttctgtatgccctgttctgtgc" 0 COMMENT "ccactcgacgtcttccatttga" 0 1248 1895 648 0 COMMENT 0) 0) 1 2 1 0 182) COMMENT (CObjectList COMMENT #155=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Contains region of the molecule from 1248 to 1895" COMMENT 1) COMMENT #156=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Tm: 81.7 C TaOpt: 59.2 C GC: 55.7" 1) COMMENT #157=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #158=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #159=(COligo "" COMMENT "TCTTCTGTATGCCCTGTTCTGTGC" COMMENT "Sense primer for PCR amplification of a region of Ple104 (pEMS1084)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " TCTTCTGTATGCCCTGTTCTGTGC" 1)) COMMENT #160=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #161=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 24 Tm: 57.2 C GC: 50.0" 1) COMMENT #162=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -173.5 kcal/mol dS: -446.3 cal/mol dG: -38.7 kcal/mol" COMMENT 1) COMMENT #163=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #164=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #165=(COligo "" "CCACTCGACGTCTTCCATTTGA" COMMENT "Antisense primer for PCR amplification of a region of Ple104 (pEMS1084)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " CCACTCGACGTCTTCCATTTGA" 1)) COMMENT #166=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #167=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.4 C GC: 50.0" 1) COMMENT #168=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -162.2 kcal/mol dS: -415.2 cal/mol dG: -36.6 kcal/mol" COMMENT 1) COMMENT #169=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 0.8" 1) COMMENT #170=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 0.0" 1))) COMMENT "#3: Product of length 648 (rating: 171)")) COMMENT #171=(CPCRFolderPar COMMENT (CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 0 COMMENT (20 COMMENT #172=(CPCRData (CStringList) (CStringList) COMMENT "cttctgtatgccctgttctgtgct" 0 COMMENT "ccactcgacgtcttccatttga" 0 1249 1895 647 0 COMMENT 0) 0) 1 2 1 0 182) COMMENT (CObjectList COMMENT #173=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Contains region of the molecule from 1249 to 1895" COMMENT 1) COMMENT #174=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Tm: 81.7 C TaOpt: 59.2 C GC: 55.8" 1) COMMENT #175=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #176=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #177=(COligo "" COMMENT "CTTCTGTATGCCCTGTTCTGTGCT" COMMENT "Sense primer for PCR amplification of a region of Ple104 (pEMS1084)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " CTTCTGTATGCCCTGTTCTGTGCT" 1)) COMMENT #178=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #179=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 24 Tm: 56.5 C GC: 50.0" 1) COMMENT #180=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -175.7 kcal/mol dS: -453.6 cal/mol dG: -38.7 kcal/mol" COMMENT 1) COMMENT #181=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #182=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #183=(COligo "" "CCACTCGACGTCTTCCATTTGA" COMMENT "Antisense primer for PCR amplification of a region of Ple104 (pEMS1084)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " CCACTCGACGTCTTCCATTTGA" 1)) COMMENT #184=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #185=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.4 C GC: 50.0" 1) COMMENT #186=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -162.2 kcal/mol dS: -415.2 cal/mol dG: -36.6 kcal/mol" COMMENT 1) COMMENT #187=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 0.0" 1) COMMENT #188=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 0.0" 1))) COMMENT "#4: Product of length 647 (rating: 171)")))) COMMENT "PCR Analysis")))) COMMENT (CGraphView COMMENT (CWStyleSheet COMMENT (CObjectList COMMENT #189=(CWidgetStyle "RSite Label" 1 (LOGPEN 0 0 13408563) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Georgia") 0.555556 0 1 5 COMMENT "@N (@S)" 0) COMMENT #190=(CWidgetStyle "Signal Label" 1 (LOGPEN 0 0 0) 1 0 1 COMMENT (LOGFONT 0 0 0 0 700 0 0 0 0 3 2 1 34 "Arial") 0.666667 0 1 1 COMMENT "@N" 0) COMMENT #191=(CWidgetStyle "Molecule Label 2" 0 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Georgia") 0.555556 0 1 16 COMMENT "@L bp" 0) COMMENT #192=(CWidgetStyle "Molecule Label 1" 0 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Verdana") 0.833333 0 1 1 COMMENT "@N" 0) COMMENT #193=(CWidgetStyle "Shape 3" 1 (LOGPEN 0 0 3355545) 1 1 COMMENT (LOGBRUSH 0 6724095 0) 0 0 1 (LOGSHAPE 9 1 0.8 1.8 0)) COMMENT #194=(CWidgetStyle "Shape 1" 1 (LOGPEN 0 0 6723840) 1 1 COMMENT (LOGBRUSH 0 10079334 0) 0 0 1 (LOGSHAPE 9 1 0.8 1.8 0)) COMMENT #195=(CWidgetStyle "Axis" 1 (LOGPEN 0 0 10079436) 2 1 COMMENT (LOGBRUSH 0 13434879 0) 0 0 1 (LOGSHAPE 10 1 0 0 0)) COMMENT #196=(CWidgetStyle "Line 2" 1 (LOGPEN 0 0 6723840) 8 0 0 0 1 COMMENT (LOGSHAPE 1 1.9 0 0 0)) COMMENT #197=(CWidgetStyle "RSite" 1 (LOGPEN 0 0 10053171) 8 0 0 0 1 COMMENT (LOGSHAPE 1 1.9 0 0 0)) COMMENT #198=(CWidgetStyle "Short Signal" 1 (LOGPEN 0 0 13395507) 10 0 0 0 1 COMMENT (LOGSHAPE 1 1.9 0 0 0)) COMMENT #199=(CWidgetStyle "Uniq RSite Label" 1 (LOGPEN 0 0 153) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Georgia") 0.555556 128 1 COMMENT 5 "@N (@S)" 0) COMMENT #200=(CWidgetStyle "Vanilla" 1 (LOGPEN 0 0 0) 1 1 COMMENT (LOGBRUSH 0 16777215 0) 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 7 48 2 18 "Times New Roman") 0.8 0 COMMENT 1 2 "?" 0) COMMENT #201=(CWidgetStyle "Mark 1" 0 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 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#210=(CWidgetStyle "Line 3" 1 (LOGPEN 0 0 16711680) 8 0 0 0 0) COMMENT #211=(CWidgetStyle "Line 1" 1 (LOGPEN 0 0 16711680) 1 0 0 0 0) COMMENT #212=(CWidgetStyle "Short Promoter" 1 (LOGPEN 0 0 128) 6 0 0 0 0) COMMENT #213=(CWidgetStyle "Motif" 1 (LOGPEN 0 0 0) 1 0 0 0 0) COMMENT #214=(CWidgetStyle "Line 0" 1 (LOGPEN 0 0 0) 8 0 0 0 0) COMMENT #215=(CWidgetStyle "Void" 0 0 0 0 0) COMMENT #216=(CWidgetStyle "General Label" 1 (LOGPEN 0 0 0) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Times New Roman") 0.91 0 COMMENT 1 3 "@T @N " 0) COMMENT #217=(CWidgetStyle "Position" 1 (LOGPEN 0 0 0) 1 0 0 0 0) COMMENT #218=(CWidgetStyle "Annotation" 0 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Times New Roman") 0.91 0 COMMENT 0 0) COMMENT #219=(CWidgetStyle "Position Label" 1 (LOGPEN 0 0 8388608) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 1 0 0 3 2 1 34 "Arial") 0.63 8388608 1 1 COMMENT "@N" 0) COMMENT #220=(CWidgetStyle "Range" 1 (LOGPEN 0 0 0) 1 1 COMMENT (LOGBRUSH 0 16777215 0) 0 0 0) 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8421376) 1 0 1 COMMENT (LOGFONT 0 0 0 0 700 255 0 0 0 3 2 1 34 "Arial") 0.833333 255 1 COMMENT 5 "@N (@S)" 0) COMMENT #228=(CWidgetStyle "Label 4" 1 (LOGPEN 0 0 8437824) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Arial") 0.722222 0 1 5 COMMENT "@N (@S)" 0)) 0.164644 1.74233 0.164644 2.53336 COMMENT (70 (CShapeMapEntry 0 "Unique RSite" 1 "Uniq RSite Label") 67 COMMENT (CShapeMapEntry 0 "ORF" 0 "ORF Label")) 40.0378 40.0378 39 39 0.1 COMMENT -17485) 1 1 0 1 1 COMMENT (mapper: 18.1183 -14.5217 40.0378 40.0378 0.01 10 -1 19808 19808 1 0 0) COMMENT #229=(CGroupWidget (CWidget 0 (0 0) 1 2 0 0 Nil -387 100) COMMENT (CObjectList COMMENT #230=(CGroupWidget (CWidget 1 (0 0) 1 2 0 0 Nil -604 100) COMMENT (CObjectList COMMENT #231=(CAxis COMMENT (CWideLine COMMENT (CWidget 0 (0 0) 1 2 0 0 #195# 41645372 0) COMMENT (LOGPEN 0 0 10079436) 2 (LOGBRUSH 0 13434879 0) 1 COMMENT 6.2745 6.2725 1 0.0214037) 0.03167) COMMENT #232=(CLabel (CWidget 1001 (0 0) 1 2 0 0 #192# 194 100) COMMENT (LOGPEN 0 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#277=(AnlzFontBackColorItem 1843 1872 43633766) COMMENT #278=(AnlzFontBackColorItem 1822 1842 40304639) COMMENT #279=(AnlzFontBackColorItem 1895 2032 43633766) COMMENT #280=(AnlzFontBackColorItem 1873 1894 40304639) COMMENT #281=(AnlzFontBackColorItem 2055 2487 43633766) COMMENT #282=(AnlzFontBackColorItem 2033 2054 40304639) COMMENT #283=(AnlzFontBackColorItem 159 1243 50318387) COMMENT #284=(AnlzFontBackColorItem 1364 1395 50318387) COMMENT #285=(AnlzFontBackColorItem 1344 1363 40304639) COMMENT #286=(AnlzFontBackColorItem 1416 1610 50318387) COMMENT #287=(AnlzFontBackColorItem 1396 1415 40304639) COMMENT #288=(AnlzFontBackColorItem 1268 1343 50318387) COMMENT #289=(AnlzFontBackColorItem 1244 1267 40304639))) (CObList) 543453793 COMMENT (CStringList) 1380863812 825242196 (CObList))) FEATURES Location/Qualifiers misc_feature 17494..137 /vntifkey="21" /label=pBS246\vector misc_recomb 13670..17481 /vntifkey="86" /label=5'\junction /note="The 5 HPRT probe anneals to genomic sequence ~560bp upstream of the BamHI site at 9263" misc_marker 18801..19801 /vntifkey="22" /label=AmpR misc_marker 12198..13644 /vntifkey="22" /label=AmpR misc_recomb 9312..12202 /vntifkey="86" /label=HPRT\3'\homology /note="Consists of part of Intron 2, all of Exon 3, and part of Intron 3" misc_recomb 4605..9311 /vntifkey="86" /label=HPRT\complementary\sequence /note="Consists of human HPRT promoter, human Exon1, Intron1, Exon2, and part of Intron2" misc_feature 2497..2530 /vntifkey="21" /label=FRT\(F5\mutant) misc_feature 2531..2542 /vntifkey="21" /label=Kozak\consensus misc_feature 2543..3259 /vntifkey="21" /label=eGFP\(+mutated\stop) misc_feature 3260..4285 /vntifkey="21" /label=Cre misc_feature 4286..4358 /vntifkey="21" /label=NLS misc_feature 4362..4395 /vntifkey="21" /label=FRT misc_feature 4549..4599 /vntifkey="21" /label=SV40\early\polyA\signal misc_feature 160..2488 /vntifkey="21" /label=Ple104\(HAP1-B) misc_feature 1612..2488 /vntifkey="21" /label=Prom\region primer 1874..1895 /vntifkey="27" /label=oEMS2380 primer 1345..1364 /vntifkey="27" /label=oEMS2400 /note="56.3" BASE COUNT 5132 a 4564 c 4689 g 5422 t 1 others ORIGIN 1 cggccgcacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc 61 acgaggccct ttcgtcttca agaattaatt ccgatcatat tcaataaccc ttaatataac 121 ttcgtataat gtatgcgaat tcgagctcct aggccggcca agttaaaggg gaacctctga 181 gggcctaggc cctcaagtcc cccattctct gctgatgtaa ggagggactg tacctgggtt 241 ctggggtggg ggtgagggct atggcggcca ctgcctggtt ctggagggga aggcctggat 301 cagagatggg ggagggcccc cgaaaggcct gcaggagaca gcaggactgc ggcgggggtg 361 aggaggtctc ccctgttcct agttcagggt gcccctccac cctgctcccc tgcagaaagt 421 aaccccggtg cctgtccccc ccaggtccct ggcaccgacc cttaggggct ctttctatta 481 cctaccctgt tccaggtgtt ttttttccca gggtcaggcc aggggagggg gcaggaggag 541 gggcctgagc cacctgtgtg ctgcccaggg ctgggcactg gagtgaatgt gccccaggtt 601 cagggggtgt gcacaattag gtggtgggtg gtctggcttt gcagcctggc aagtcttctg 661 caccctccca tcctctttcg gatccaggag ggtccatgga tgcacactgg gagggaggac 721 aggaggccaa gggtctttgc tgtgtctgat ttctccccct agatgagaag caggagaggg 781 agggggtggt tgacagagga aaaccacttg agaggcctgg atcagagact acccccagtc 841 tcctggacag ccctaggctg tcgaaaggca gatatctgtc agccaggcca aaatgcttac 901 ttccagaatg acgggccctg agccctggtt tctccatttg taaaatggag tctctgtccc 961 tactccactt tctgtggggc gactgagcct gtaggtgagg aagccacgtg gggaggtata 1021 cgggctgaca ctgccctctg gtggccgtag ccagagttgc tggctgccag tgcgctgacc 1081 tagagggtgt ggtcagctgg gggagctttc agggtctcct aacttcatct ctgtcttctc 1141 catcccactg cctctgcctt cttccccgtc aaactctcca ggggtttccc agggcttctt 1201 cctcatgggg ccagaaacaa tttaattcag caggagtgta ataggcttct tctgtatgcc 1261 ctgttctgtg cttggtggtt ggggctgcag agatgtccaa ggcacagaga gacagtccag 1321 gaagctgggc tgtgggagga ccactcggca gagcctatgg agctgtagcc tcacggaagc 1381 ctgggctgtg ccgatgcccg aacagccacc tccttatgtt cttcaacctc actgtcgtag 1441 caaaccctgg ggttgtgtgt ctccttcaag ctttctgctg acaaagaaac cataggaaac 1501 atgcagactt tgggctgggc acagtggctc acgcctgtaa tcccagcact ttgggagacc 1561 aagggggagg cggatcacgt gaggtcggga gttcgagacc agcctaacca acacacgcca 1621 ccatgcttgg ctaatttttg tatttttagt agagacaggg tttcaccagg tggccaggct 1681 tgtctggaac tcctgacctc aggtgattcg cctgccccgg cctcccaaag tgctgggatt 1741 acaggcgtga gccactgcgc ccggctacag ggagactttt aagaggcgtc tcccaaggag 1801 aacaggggtt cgccagcatg gcaagaggag ggcaaagcga actagggttg gaggtgaaga 1861 acagttaatc caatcaaatg gaagacgtcg agtggcgcga gcactggggt gtgagcgcga 1921 gggtcagggt tgcagcgcca ggcagggttg cagcgccagg cagggttgca gcgccaggca 1981 gggttgcagc gccaggcagg gttgcagcac gttggaagcg cactggagtt tggaatgtct 2041 ctcccagtca accgggaaag acgcagcgct gccctaatgc actggccacc ccagaccgta 2101 tgcacaacgt tgggttggca gcttctacac cagggaaggc acccgaaacc ccggcgcggc 2161 agctggggag gggctgcccc cagccaggga cccgcgagtg ctggggcagc cgcggctttg 2221 ggcagaacca ttctccggag atggagaggg cagaaaaaat ctttcgctcc catcccaact 2281 cggagccatc cggaacttgc actcgccctt cttgccctgc ttttcttact gtgctctgtt 2341 ccgtcaccac tagctcccct cccgcgcggc tctgagtccc gcttgccact agttgcttct 2401 cggcagcccc cacccctacg cgtggtccgg cccggcggct ggcgcaggtt ataaaggtcc 2461 cgccgcagca gcggacggca gactcgaggg cgcgccgaag ttcctattct tcaaaaggta 2521 taggaacttc gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 2581 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 2641 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 2701 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 2761 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 2821 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 2881 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 2941 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 3001 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 3061 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 3121 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 3181 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 3241 ggacgagctg tacaagttat ccaatttact gaccgtacac caaaatttgc ctgcattacc 3301 ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca 3361 ggcgttttct gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg 3421 gtgcaagttg aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct 3481 tctatatctt caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct 3541 aaacatgctt catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact 3601 ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct 3661 agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg 3721 ccaggatata cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc 3781 cgaaattgcc aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat 3841 ccatattggc agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct 3901 gggggtaact aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa 3961 taactacctg ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca 4021 gctatcaact cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc 4081 taaggatgac tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc 4141 cgcgcgagat atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg 4201 gaccaatgta aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat 4261 ggtgcgcctg ctggaagatg gcgatgatcc aaaaaagaag agaaaggtag atccaaaaaa 4321 gaagagaaag gtagatccaa aaaagaagag aaaggtaata agaagttcct attctctaga 4381 aagtatagga acttcagcgg ccgcgactct agatcataat cagccatacc acatttgtag 4441 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 4501 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4561 gcatcacaaa tttcacaaat aaagcatttt tttcactgcg aattctcgta aaactcttca 4621 tggcagtagt tattattctc tctctctctc tttttctttt ttcttgagac aggatatttc 4681 tctgttgccc aggctggagt gcagtggcac agtcttggct cactgcagcc tggacctcct 4741 gggctcaagc catcctccca cctcagcctc ccaagtagct ggggctacag gcacatggcc 4801 accaggccag ataatttttc atttttgtag agactgagtc tcaccatgtt acccaggttt 4861 attattctca ttttttagat gaagagactg aggtccagag aagctcaatg acttgcctag 4921 ttttacaaat ctcctgccat cacatacccc tcagcgtcct taataagagg gaggccacca 4981 actatgtgct gggcactgtg gtggatgctg gagctatagg gttgagtata taagaaatgg 5041 tgttgctgga gcaactgttg cttgcttacc tgacctatct gagaattaat tagcagggga 5101 acatattttt gttttcagat tcaatataag aacttgtgtg ggcaaaaata aagatcagta 5161 gtaataacag tagttcccat ttgctgactg tactgtccta agtgcatata tatatacata 5221 cacacacgca tacctatact cctctaatac tcaaaatgat cctgtttatg tattgttaat 5281 atgctcattt tatttttaaa tttttattta tttttatttt tatttatttt tgagacggag 5341 tctcattctg tcgcggaggc tgaagtgcag tggtgcgatc tcagctcagt gcgacctccg 5401 cctcccgggt tcaagtgatt ctcctgcctc agctccggac tagctgggat tacaggcgcc 5461 cgcctccacg cccagctaat ttttgtattt ttagtagaga tggggtttcg ccatgttggc 5521 caggctggtc tcgtactcct gaccttgagt gatccacctg cctcggcctc ccaaagtgct 5581 gggattacag gcatgagcca ccgcgccggg ctaatatgct cattttagtg aggcaaaaat 5641 agaggctcag agtctgattt gtacaaaact acagagcagt taagtgtcct ctcagatgtg 5701 taccctgatc tgggtgactc taggactcta ggtctcaact gttacaacca gttaagggtt 5761 tggggaagca ctgggccaag agtcaggaaa atggaagcca caggtagtgc aaggtcttgg 5821 gaatgggacg tctggtccaa ggattcacgc gatgactgga acccgaagag ccggggcccg 5881 gtttacggcc gccatgaagc aacgcgcgcc ggtaggtttg ggaatcaggg agccctctga 5941 ataggagact gagttgggag ggaaaggggc ttcgctgggg gagcctcggc ttcttctggg 6001 agaaaattcc cacggctacc tagtgagcct gcaaactggt aggcgccggc gtaggcgcgc 6061 gggcggggcc gggggcgggg cctgcggggc gtggcggggc gggcagaggg cggggcctgc 6121 ttctcctcag cttcaggcgg ctgcgacgag ccctcaggcg aacctctcgg ctttcccgcg 6181 cggcgccgcc tcttgctgcg cctccgcctc ctcctctgct ccgccaccgg cttcctcctc 6241 ctgagcagtc agcccgcgcg ccggccggct ccgttatggc gacccgcagc cctggcgtcg 6301 tggtgagcag ctcggcctgc cggccctggc cggttcaggc ccacgcggca ggtggcggcc 6361 gggccctgag gcgcgggatc gatccatgac acccccccgc cccccatatc tcactataat 6421 tcactgtcaa atggctttta tgatttggag tttggggata cttttaaaga cagggtcttg 6481 gtagctccta gatggcttgg gattcactta gtaacccttg ctggcctaaa attcaggaac 6541 ttctacctca tcttcccaag tactgggtta gtagggatgt ccagcatatt tagcttgctt 6601 taatgatttt gctgtacttt tttctgtgtg tacaggtgca tgtatattta taaaactaac 6661 ataatatatt ttacccccaa agtaccataa ccctaattaa tgctattaaa tgttttgtat 6721 gctgggctct gggcatacta atataaccaa tattatacaa tgcaaaacaa ttttggaata 6781 tcttctgtaa aattatctta ttagtaagtt gtaaccatct cagacttcta agtttgagtt 6841 tgtaaaggat cagttttttt tggtgaccta cacacacaca cacacacaca cacacacaca 6901 cacacacagg ggggtggatt ctgagaattt agagaattta gatggttaat accagttttt 6961 atttttattc tgtgtgatat ttatttatta agatttattt atttattaca tgtaggtaca 7021 ctgtatctgt cttcagacac accagaagag ggcatcagat ctcgttacag atggttgtga 7081 gtcaccatgt ggttgctggg atttgaactc aggacctctg gaagaacagt cagtgctctt 7141 aaccgttgag ccatctcacc agccccgtga tattttattt aacttactag aatatactgt 7201 gttaaaacat attttagggg attggggaga tggcccattc tgtaaagtgt ctgccatgtc 7261 atcttcagga cctgcgtttg taaccccagc atggagaaga cagagacagg gagacccctg 7321 ggcttcctgg caagctagac tagcaaattt ggttaactag gttcaatgag agactcagtc 7381 tcaaaacata agaagtgaat tttttagacc caatttcaat ctctgaactc catacgtatg 7441 cacatgtacc tgcatataaa tatatataca cacccataaa cacatgcaca cacaaaaaat 7501 ggttcaaaag ttcttaaaat gtataagagt agaaagtgta gagccgggag tggtggcgca 7561 cgcctttaat cccagcactt gggagcagag gcaggtggat ttctgagttc gaggccagcc 7621 tggtctacag agtgagttcc aggacagcca gagctataca gagaaatcct gcctcaaaaa 7681 accaaaaata aagtgtatat actaatcttt catgcctaaa atatttattt ttgccacatt 7741 tttcatacct ctttaatatt tttctttttc tctttgtgct tcaaaggaaa ttttgtgtca 7801 tgttttactt ttcattcttg actctattca gcagtaagac gcagcatggc tttcttagag 7861 aagtataatc tattacaaaa cggtaacatt taactgctct acataccctc ttgtagctaa 7921 attcctctgt taaactaaga cttattatct aaacgtaatt aaatgttgat atcttataat 7981 gctccacttt gaaacagctg tctttctttt gcagattagc gatgatgaac caggttatga 8041 cctagatttg ttttgtatac ctaatcatta tgccgaggat ttggaaaaag tgtttattcc 8101 tcatggactg attatggaca ggttagtaag acctcgattg aagttatttt tagagggtgt 8161 gtgtgtggtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtaag agagagagga 8221 gaggcatgtg tgatatatat gtgaagtgta ttgcatgtac atggaatcca gagaaggaca 8281 ttaggtgtcc tactctgttg ctctcttatt cccttgaatc agtgtctctt gctgaacctg 8341 gggcatacat actgatttgg ctaggctgcc tggctagcaa gacccaggga tctgcttgtc 8401 tctgcccccc agtgctgacg ttacaagcag gtgcaaccac atctggtttt tatatgggta 8461 ctgggggatc tgaacttagg gcctgatgcg tgcccaagtg ctctcacccg ccaagccacc 8521 tctccagcca aagggttttt tttcttgtat taattctaac ttagcaactg ttgagtagtt 8581 gatgtttgaa tataagctaa gccaacttat tagaataaat gttttctatt ttttacttgt 8641 atgtgtgagt gcctgcatgt atgtatgtga agcatatgta ttccaggatg tcagaattgt 8701 gatggattcc ctggaactgg agttacagac cattgtgaac cactgtgtat gggtgctggg 8761 aactgaactt taggtcctct gcaaaaacag taaggatctt aactgataaa ctatctttct 8821 agccctaagg ctttctactt ttaaagttcg tttctctttc tctctctctc tctctctctc 8881 tctctctctc tccctctccc tccctcctcc tcctcctcct ctttcccccc tctcgcctcc 8941 tctttctctc tcccctccct ctctctctct cacacacaca cacagacaga gagaggagag 9001 agaaaggaag ccttactaat taattaccta gttttagaca taactagtgg gaatccatag 9061 taagtaaatc aacattcttg aagagaggtt agaagaattg gctctcgttg tccatcgagg 9121 ctgtataaga ttattcttct gttttctcag cttaccatac atattttcta cgtagacaat 9181 accaattcaa atacactgta cgtaaagtat atgcgcacac acacacacac acacacacac 9241 acacacacac aagttacata ttgtcatata gtactaaaat ttatggactt tttcatatta 9301 atacatctag agctatcatg ttttttatct tttaaaaatt actttacggg gctacagaga 9361 tgtctcagca gttaggagca ctgactgctc ttccagaggt cctgagttca attcccagca 9421 accacatggt ggctcacaac catctgtaat gggatctgat ccactcttat ggtgtgtctg 9481 aagacagtga cagtgtactc atatacatga aataaatctt taaaaaatta ctttaccatc 9541 agggtatata tatatatata tatgtgtgtg tgtgtgtgtg tgtgtatgaa gtatttgcta 9601 tttgagaggg tctcactgtt tagctcagtc tggcttggaa cgttctgtat agcccagact 9661 ggcctcaact cagcagtccc cctgaaaggc tctcatatgt ggagattaca ggtatgtgct 9721 actgtgccca gccctacatg ttactttgtt tttgaaatag ggtatcaaat agcttagact 9781 ggcttctaag tcagtgtgta gctaaggatg gtctgagctt ctgaacctgc tacttcaact 9841 cctggtgtgc catcatggcc agtttgcaca gttgtgagag tagaactcag gacctcctat 9901 gtgttactag acatgttctc taccagctga gctacatccc aattcgccta catttacttt 9961 tattttttaa aagatttatt tatgtataca gcactctgcc tgcatgtaca tctgcatgcc 10021 agaagagggc accagatctc atcatacata gttgtgagcc accatgtggt tgctggggat 10081 tgaactcagg gcctctggaa gagcagtcag tgctcttaac ctctgagcca tctctgtagc 10141 cccctacatt tattttaaag attataggct attctattca tagatgtgcc ataactgatt 10201 taaccagatc ttcattattt tgtacttagt agagtatatt agagggaatg aatcttaaat 10261 acacattttg acacttgctc agctttagac aagtaaaaat cactggtcaa ggaaatggtg 10321 cttagtggat gcttctgaac tgtcaaattg tcctccaaca agttacatca ttttatactt 10381 ccaacgttat gtgagagttc atttttcttt aggctcttct agtattttat atttgagaaa 10441 taaaatttaa ccaaaatttg ccaatctagt aagcttgttt ttatttgtat ctttaaattt 10501 tatgatagat gttatagtgt actctcctct cccctcccct cccctctcct cctcagcagg 10561 atcttgttat gtagccctaa gggatctaat acttactctg tagtccaggc tgtccttcag 10621 ctcatgagac acaccatcat gcttggcatg tttcttgttc tgaaaggtat ctggttttaa 10681 ttttggttga gctggtgtgt tggtgtatat ctgtaatctc agcacaggca agtagagaca 10741 aggattatga ttacagtcca gcctgggcta tttagaggga ttctctttaa tctttcctca 10801 tgccccaaaa tcttaccttt ggtatatgaa aaatagtctc cacttctgca aaatattgct 10861 ttatgaagta agaattaatt cccttcatag agacaaggaa tgtgtcctgt aaaagtttaa 10921 tgtgtaagaa gtatttgtta taaaagataa atattcagaa tcttcttttt aattcctgat 10981 tttatttcta taggactgaa agacttgctc gagatgtcat gaaggagatg ggaggccatc 11041 acattgtggc cctctgtgtg ctcaaggggg gctataagtt ctttgctgac ctgctggatt 11101 acattaaagc actgaataga aatagtgata gatccattcc tatgactgta gattttatca 11161 gactgaagag ctactgtgta agtataatta acttataatt aaaaaaatag ggccattcta 11221 gttttatcta tatttttttt aaacttgtgc aaactatgct acataatcaa cttacttgat 11281 tatgtaatat atatagtaat agttggaatt actattatta catatttatt tttctgtgct 11341 ggggtagaac ccagggaact aagtactgta ataaccgagc ttcattcatc ctagtcattt 11401 ttaatttttt tttttttttt tgttttttcg agacaaggtt tctctgtgta gccctggctg 11461 tcctggaact cactctgtag accaggctgg cctcgaactc agaaatctgc ctgcctctgc 11521 ctcccagtgc tgagattaat gacgtgcacc accactgcct ggcatttttt taaagtttta 11581 atttgagacc ctttctcact aacttgctca cactatcctc gaactcatta cgtgggctgg 11641 tctagaaatt tcagtcaact tgcctcagct tctcaagtag ctgggattac aggcaggtgc 11701 taccacttct ggccatgatc attgttgttg ttgctattat tattattatt atttcactga 11761 tacttcatat cacaacacag aatagtaact gtagttcatg accaacaaaa aagttgtaca 11821 gtttctctgt tgtcttattt tgatctggat tttgatttgt ttcatggcac tgggcaggct 11881 aatctcaaac tgcatggctc aaatagtcct tctgtcttaa tctcccatgt tctggaagta 11941 taggtatcac tacacctagc tgtattttca tattttaaaa aaaacgccta agaattgcta 12001 ttgaatgagg cggagagatg gctcagcagt cacaagtact tgtagaggac tggggtttgg 12061 ttactagcac ctcaaagatg gcttacaatt atctgtagct ccaattccag aggatctgtg 12121 tggtaaagct gcatgtatat gggcatgcag gtaaaactgc taaacatata aaaatagctc 12181 agtcttaaga gaattggtcg acgtaagctc attcgagtcg actttttcgc cctttgacgt 12241 tggagtccac gttctttaat agtgdgactc ttgttccaaa ctggaacaac actcaaccct 12301 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 12361 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt 12421 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 12481 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 12541 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 12601 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 12661 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 12721 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 12781 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 12841 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 12901 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 12961 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 13021 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 13081 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 13141 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 13201 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 13261 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 13321 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 13381 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 13441 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 13501 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 13561 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 13621 aaacaaaaaa accaccgcta ccaggtcgag tagggataac agggtaatgg atcctcgttg 13681 ggcacctgca ctcatgtaca caggcccaca tatagatgta cacacgcata tcattcaaaa 13741 tagaaattat ttatacaagt agtgcgttgg agagctggct caggagttgt taatgcttgc 13801 tacccttgca aaggacccaa gttcaatttc cagcacctgt gttgggcatc tcacaaccac 13861 atgtaaatct agtttcaggg gatttcaccc ctcttctgaa ctcttcaggc acttataccc 13921 acaaatggac atctccacat agagacacat acttttaaaa aagtaatcta gggtctagag 13981 aaatggttat agaccacgga ctgggatcag ttcccagaag ccacatcctg tggctcacaa 14041 ttgcctgtga atcaagttct agatctggtg gcctcgtcta gcctgcagtc atcaacaaac 14101 acatgcacac atccatacac acataattta aaagaaagcg aaagtaacct ataatgctaa 14161 acaaagcaca ttctcagtgt agactgcagt ggtgatgaac cttgaaaata tattaataga 14221 agccagacac gtaagtcaca aatggtataa ctcaatttat atgaaattcc cagagagtcc 14281 aaagttaaaa aaaacaaaaa ggcagattat tggttgtcag tagctacaga gatgggacat 14341 tgaagatatt aataggtata gggttgagga ttcatggcct ttccctcttc catgttagca 14401 tgtctattgt tgttaacctt attcaactca tgttagggca atcatattga tgagacttaa 14461 tgagtatagc ttgacattcc tagaagacag agtctcacat cagactctct gatcctctga 14521 ctcttagaat ctttcaacac cctcttctga aatgatctct gagcctttgg ttgtgactga 14581 attatgagaa agaattttat gttgaatgaa ttttacttca gcatttttac aaaactgaag 14641 ctggggatgg cgtttcatgc gtgcagtccc taccatttgc ctataagcct ggaggaggca 14701 gaagtagaat taggcttttc agggacatct tcagctgcat catgagcttg aggacaactt 14761 gagctacata agattcccat ccccaccccc acacacacac acacatctag tatgtaccta 14821 gtatgcagtc ttttcatatg tttaaaatgt agaaaggtat atgtgaagtc tcccttttcc 14881 ccacaactta gtccctccaa agtattgcat cagtttcctt gtacatgatt ttatgcatat 14941 ataagataca ctatttctct ctctctctct ctccttccct ccttccctcc aacctctggg 15001 tatgagttga gggttatgta tgtttcaagt tgtaataaga attgccactc aaacatttca 15061 tcccatttta agttaagttt gtctgatttt ccactttaga agggcagcca tgatggctca 15121 tgcctataat cccatcctag cacttgtaag cctaatacat aagactggag gattgtcatc 15181 agcttgaggc tagcctggtc tacctaatga gttccagcca gccaagagta gaatataaga 15241 ccctgtctcc aaaaaatgta ccaaaggggc tggaggggta actcagtggt taagaacagt 15301 agctgctctt ccagaggacc tgggttcaat tcccagcacc cacatggcag cttgcaacta 15361 tctgtaactg tccagctttg ggatatccga caccattata ccaattacat aaaatgaaaa 15421 aattgtaaaa tgtgctaaaa tgagtgggga gggggagcat ggtgtggttc tgtgggcttg 15481 ggattttgcc atgaatccat gactatatct cagcaatatg tggattattt ctggaaacct 15541 gtggggcatg ggtgtatggg aagaggggag agagagagag cccgcatcca gccagagttc 15601 ctgtgctctg ggcaggcgga cgtgggagga ctgctggacg atttccactt ggccctgggt 15661 gggcatctgg ctgtgtgaag ccactgaccc cactcagcgg ggggtagata aggggcattt 15721 cccataccag gttctcagac tccagcattc cacgctggat acagcagaag agctgagaac 15781 agaatagggg ccccagggag acactctcgt cccagagata aagaaggcag agggagagag 15841 attcctacgc aggcgagagg ccttagtctg ggccttgatg aacagagacg gtctatggtt 15901 ttagagcttt attgtagaaa ggcaggggga aagagagaag gtagaaagag agagaccagc 15961 catggccaag aggagagaag gggggaaagg agagagagaa gaaaggctag agagtaagaa 16021 aggttaaaga gacagaagta aaagcttaga gtaagagaga gagagagagc aagagggaga 16081 gtaagaaaag taaaagctta agagagcgag gaggggccag cccctctaat agtgggcttg 16141 ttatcttgtt gttgctaggt aactgtgaag cgtctagcct gaaggccaga agcttgggac 16201 attgtctatg tgactgaaag ccatgagcct ctactgtggg ggctgtggag gtggtaactt 16261 ggacaggagc caggggttcc aggagacatg agggaatgcc ttctgtccca tgtaggtagg 16321 aattaccacc accggctttc accgtagttc actggagact gtttgtacat agcccattgc 16381 cccacagaaa ccaggcactc actgtttcat ttgcataaga attagaaggc tttattttgg 16441 ctgggattgg tgacttgccc cagaaattcc agcttttggg aagccgaatc ctggagaacc 16501 aggagttcaa ggtcatcctc aggtacctca tttgttcaag accagcctgt gctaccatga 16561 gaccttgtcc ctactcccct caaccccaaa acacgctgtt ttcattttat aggttgtact 16621 aaccatggag gatggtcatt tgcctttctc ttgatttcag aagcgtagcg cgggctttgc 16681 accatgatct agtgtttcca gataggcttg cctctccaca gccaggctta tcctgcttca 16741 gtcccatgtt tggcaagggc aacatatgac tgaagaactt gcatagcctt atggtgtatc 16801 atgactctta gccccagttt ataactgtct aacagctggt ggcttttgtg aaaaaggaaa 16861 gcatcatttg aacttttaac ttttcagctg tttgttttga ctttgcataa atactgagac 16921 tagctaggaa atgagctttt gtttcatttg ctaaaaatga tttttttttc atgaggcaat 16981 ttcaagcaat ttgaatttag caatctctac ttgtctgaaa agaagttatt aattaaaaaa 17041 aaacaaacaa acaggaacct attatgctgg ctagtcacat gaaagaatcc aaatgaacac 17101 agtcaaggct gaatcaacac atgaaattat tttctaattc cccaaattac atttatgcta 17161 attagtatgt agtaaaacat tttaattgtt tccaatgtta aacattacag aaataatttt 17221 gatgttttcc taataaccca gcctttgatc atgtttgcat ttatttgatt gttgtcagtt 17281 tattaaataa ttaagtaaga gcaacaagat aaaatgactt catgagagtc cttttctgta 17341 acttagaacg tcagtagtca taggaactgc ggtcgttccc ttacctttca gaagtaggct 17401 aaagagttga acgcaaaggt ggtttttcga atcttcctga tccttaagcc cccactaggt 17461 ttatattcta tatttgatat cggattcgaa cccctcgagc ttaggatccg gaacccttaa 17521 tataactaag ttattaggtc cctcgaagag gttcactagt actggccatt gcggccgctt 17581 cgaggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 17641 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 17701 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga 17761 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc 17821 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt 17881 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 17941 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 18001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 18061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 18121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 18181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 18241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 18301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 18361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 18421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 18481 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 18541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 18601 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 18661 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 18721 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 18781 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 18841 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 18901 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 18961 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 19021 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 19081 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg 19141 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 19201 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 19261 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 19321 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 19381 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata 19441 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 19501 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 19561 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 19621 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 19681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 19741 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 19801 ctgacgtg //