LOCUS Ple219\(pEMS1182 18291 bp DNA circular 8-OCT-2010 SOURCE ORGANISM COMMENT The sequence shown is the expected sequence. Sequence verification of actual construct was not available PCR genotyping: Sense primer: oEMS2646 - 5' CTCCTCTTACACACCCAACTCCCT - 3' Tm=56.6 Deg C Antisense primer: oEMS2622 - 5' GACAGTAGCCACACACCCAGACA - 3 Tm=56.3 Deg C Cycling conditions: Step 1: 94 Deg C for 3 min Step 2: 94 Deg C for 1 min Step 3: 61 Deg C for 1 min Step 4: 72 Deg C for 45 sec Step 5: repeat Step 2-4 34 times Step 6: 72 Deg C for 5 min 535 bp product COMMENT This file is created by Vector NTI http://www.invitrogen.com/ COMMENT ORIGDB|GenBank COMMENT VNTDATE|567533049| COMMENT VNTDBDATE|567533131| COMMENT LSOWNER| COMMENT VNTNAME|Ple219 (pEMS1182)| COMMENT VNTAUTHORNAME|Russell Bonaguro| COMMENT Vector_NTI_Display_Data_(Do_Not_Edit!) COMMENT (SXF COMMENT (CGexDoc "Ple219 (pEMS1182)" 0 18291 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 100 COMMENT 0 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 18291 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) "5' junction" 86 0 0 12153 15964 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "12153..15964") COMMENT #1=(CFSignal (CObList) "AmpR" 22 0 0 17284 18284 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "17284..18284") COMMENT #2=(CFSignal (CObList) "AmpR" 22 0 0 10681 12127 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "10681..12127") COMMENT #3=(CFSignal (CObList) "HPRT 3' homology" 86 0 0 7795 10685 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "7795..10685") COMMENT #4=(CFSignal (CObList) "HPRT complementary sequence" 86 0 0 3088 7794 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "3088..7794") COMMENT #5=(CFSignal (CObList) "FRT (F5 mutant)" 21 0 0 1970 2003 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "1970..2003") COMMENT #6=(CFSignal (CObList) "NLS" 21 0 0 2767 2839 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "2767..2839") COMMENT #7=(CFSignal (CObList) "FRT" 21 0 0 2845 2878 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "2845..2878") COMMENT #8=(CFSignal (CObList) "SV40 early polyA signal" 21 0 0 3032 3082 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "3032..3082") COMMENT #9=(CFSignal (CObList) "EGFP (mutated stop TAA -> TTA)" 21 0 0 2011 2730 COMMENT 0 (CStringList) (CStringList) 1 1 1 1 "2011..2730") COMMENT #10=(CFSignal (CObList) "Kozak consensus" 21 0 0 2004 2014 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "2004..2014") COMMENT #11=(CFSignal (CObList) "Tetracysteine residue" 21 0 0 2731 2766 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "2731..2766") COMMENT #12=(CFSignal (CObList) "I-Sce I" 21 0 0 12134 12151 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "12134..12151") COMMENT #13=(CFSignal (CObList) "TAC3-B" 21 0 0 160 1961 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "160..1961") COMMENT #14=(CFSignal (CObList) "Prom region" 21 0 0 1582 1961 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "1582..1961") COMMENT #15=(CFSignal (CObList) "oEMS2646" 27 0 0 1155 1178 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "1155..1178") COMMENT #16=(CFSignal (CObList) "oEMS2622" 27 0 0 1667 1689 0 (CStringList) COMMENT (CStringList) 1 1 1 1 "1667..1689") COMMENT #17=(CFSignal (CObList) "Regions 1, 2, 3" 21 0 0 160 1581 0 COMMENT (CStringList) (CStringList) 1 1 1 1 "")) (CObList) COMMENT (CObList COMMENT #18=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "gcccatgttctttgcccaca" 63 1369 20 0 100 100 100 57.9283 COMMENT 55 -157.5 -399.9 -36.4898 -16.4814) COMMENT (CPrimer 0 "aaaggctctgggagggtctagg" 63 1774 22 1 100 100 100 COMMENT 56.7103 59.0909 -177.2 -457.5 -39.0164 -13.2136) 171 1369 1795 427 COMMENT 80.8865 55.0351 58.7337) COMMENT #19=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "cccatgttctttgcccacaca" 63 1370 21 0 100 100 100 57.852 COMMENT 52.381 -158.7 -403.4 -36.6463 -14.684) COMMENT (CPrimer 0 "aaaggctctgggagggtctagg" 63 1774 22 1 100 100 100 COMMENT 56.7103 59.0909 -177.2 -457.5 -39.0164 -13.2136) 171 1370 1795 426 COMMENT 80.8395 54.9296 58.7008) COMMENT #20=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "atgttctttgcccacacagcc" 63 1373 21 0 100 100 100 56.2785 COMMENT 52.381 -160.8 -411.4 -36.3611 -14.3988) COMMENT (CPrimer 0 "aaaggctctgggagggtctagg" 63 1774 22 1 100 100 100 COMMENT 56.7103 59.0909 -177.2 -457.5 -39.0164 -13.2136) 171 1373 1795 423 COMMENT 80.6972 54.6099 58.4716) COMMENT #21=(CPrimers (CStringList) (CStringList) COMMENT (CPrimer 0 "gttctttgcccacacagccc" 63 1375 20 0 100 100 100 56.6814 COMMENT 60 -157.4 -401.2 -36.0022 -16.126) COMMENT (CPrimer 0 "aaaggctctgggagggtctagg" 63 1774 22 1 100 100 100 COMMENT 56.7103 59.0909 -177.2 -457.5 -39.0164 -13.2136) 171 1375 1795 421 COMMENT 80.796 54.8694 58.6616)) (CObList) (CObList) (CObList) (CObList) COMMENT (CTextView 0 COMMENT #22=(CGroupPar (CParagraph 0 (0 0) 1 2 0 0 180) COMMENT (CObjectList COMMENT #23=(CRefLinePar COMMENT (CLinePar (CParagraph 0 (0 0) 0 2 0 1 233) COMMENT "Ple219 (pEMS1182)" 2) 5 "" 0 4) COMMENT #24=(CFolderPar COMMENT (CGroupPar (CParagraph 1 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #25=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "DNA 'Ple219 (pEMS1182)'" 1) COMMENT #26=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Local object" 1) COMMENT #27=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Created: 10/08/10 04:04PM" 1) COMMENT #28=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Last Modified: 10/08/10 04:05PM" 1) COMMENT #29=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "length: 18291 bp" 1) COMMENT #30=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "storage type: Basic" 1) COMMENT #31=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "form: Circular" 1))) "General Description") COMMENT #32=(CFolderPar COMMENT (CGroupPar (CParagraph 2 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #33=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Original Source Database: GenBank" 1) COMMENT #34=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Modification Date in the Original DB: 5-JUL-2007" 1))) COMMENT "Standard Fields") COMMENT #35=(CRefLinePar COMMENT (CLinePar (CParagraph 0 (0 0) 0 2 0 0 233) "Comments" 2) 1 "" COMMENT 0 0) COMMENT #36=(CFolderPar COMMENT (CGroupPar (CParagraph 8 (0 0) 1 2 0 0 178) (CObjectList)) COMMENT "Annotations") COMMENT #37=(CFolderPar COMMENT (CGroupPar (CParagraph 12 (6 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #38=(CFolderPar COMMENT (CGroupPar (CParagraph 21 (7 21 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #39=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 160 (3 #13# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #40=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 160 End: 1961" 1) COMMENT #41=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #42=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 160..1961" 1))) "TAC3-B") COMMENT #43=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 160 (3 #17# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #44=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 160 End: 1581" 1))) COMMENT "Regions 1, 2, 3") COMMENT #45=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1582 (3 #14# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #46=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1582 End: 1961" 1) COMMENT #47=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #48=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1582..1961" 1))) "Prom region") COMMENT #49=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1970 (3 #5# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #50=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1970 End: 2003" 1) COMMENT #51=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #52=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1970..2003" 1))) "FRT (F5 mutant)") COMMENT #53=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 2004 (3 #10# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #54=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 2004 End: 2014" 1) COMMENT #55=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #56=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 2004..2014" 1))) "Kozak consensus") COMMENT #57=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 2011 (3 #9# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #58=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 2011 End: 2730" 1) COMMENT #59=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #60=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 2011..2730" 1))) COMMENT "EGFP (mutated stop TAA -> TTA)") COMMENT #61=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 2731 (3 #11# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #62=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 2731 End: 2766" 1) COMMENT #63=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #64=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 2731..2766" 1))) COMMENT "Tetracysteine residue") COMMENT #65=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 2767 (3 #6# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #66=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 2767 End: 2839" 1) COMMENT #67=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #68=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 2767..2839" 1))) "NLS") COMMENT #69=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 2845 (3 #7# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #70=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 2845 End: 2878" 1) COMMENT #71=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #72=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 2845..2878" 1))) "FRT") COMMENT #73=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 3032 (3 #8# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #74=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 3032 End: 3082" 1) COMMENT #75=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #76=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 3032..3082" 1))) COMMENT "SV40 early polyA signal") COMMENT #77=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 12134 (3 #12# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #78=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 12134 End: 12151" 1) COMMENT #79=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #80=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 12134..12151" 1))) "I-Sce I"))) COMMENT "Misc. Feature (11 total)") COMMENT #81=(CFolderPar COMMENT (CGroupPar (CParagraph 22 (7 22 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #82=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 10681 (3 #2# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #83=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 10681 End: 12127" 1) COMMENT #84=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #85=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 10681..12127" 1))) "AmpR") COMMENT #86=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 17284 (3 #1# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #87=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 17284 End: 18284" 1) COMMENT #88=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #89=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 17284..18284" 1))) "AmpR"))) COMMENT "Misc. Marker (2 total)") COMMENT #90=(CFolderPar COMMENT (CGroupPar (CParagraph 27 (7 27 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #91=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1155 (3 #15# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #92=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1155 End: 1178" 1) COMMENT #93=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) "56.6" COMMENT 1) COMMENT #94=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #95=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1155..1178" 1))) "oEMS2646") COMMENT #96=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1667 (3 #16# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #97=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1667 End: 1689" 1) COMMENT #98=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #99=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1667..1689" 1))) "oEMS2622"))) COMMENT "Primer (2 total)") COMMENT #100=(CFolderPar COMMENT (CGroupPar (CParagraph 86 (7 86 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #101=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 3088 (3 #4# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #102=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 3088 End: 7794" 1) COMMENT #103=(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 #104=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #105=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 3088..7794" 1))) COMMENT "HPRT complementary sequence") COMMENT #106=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 7795 (3 #3# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #107=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 7795 End: 10685" 1) COMMENT #108=(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 #109=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #110=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 7795..10685" 1))) COMMENT "HPRT 3' homology") COMMENT #111=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 12153 (3 #0# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #112=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 12153 End: 15964" 1) COMMENT #113=(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 #114=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #115=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 12153..15964" 1))) "5' junction"))) COMMENT "Misc. 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COMMENT " Tm: 80.9 C TaOpt: 58.7 C GC: 55.0" 1) COMMENT #124=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #125=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #126=(COligo "" "GCCCATGTTCTTTGCCCACA" COMMENT "Sense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " GCCCATGTTCTTTGCCCACA" 1)) COMMENT #127=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #128=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 20 Tm: 57.9 C GC: 55.0" 1) COMMENT #129=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -157.5 kcal/mol dS: -399.9 cal/mol dG: -36.5 kcal/mol" COMMENT 1) COMMENT #130=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #131=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #132=(COligo "" "AAAGGCTCTGGGAGGGTCTAGG" COMMENT "Antisense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " AAAGGCTCTGGGAGGGTCTAGG" 1)) COMMENT #133=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #134=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.7 C GC: 59.1" 1) COMMENT #135=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -177.2 kcal/mol dS: -457.5 cal/mol dG: -39.0 kcal/mol" COMMENT 1) COMMENT #136=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 1.2" 1) COMMENT #137=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 4.1" 1))) COMMENT "#1: Product of length 427 (rating: 171)")) COMMENT #138=(CPCRFolderPar COMMENT (CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 0 COMMENT (20 COMMENT #139=(CPCRData (CStringList) (CStringList) COMMENT "cccatgttctttgcccacaca" 0 COMMENT "aaaggctctgggagggtctagg" 0 1370 1795 426 0 COMMENT 0) 0) 1 2 1 0 182) COMMENT (CObjectList COMMENT #140=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Contains region of the molecule from 1370 to 1795" COMMENT 1) COMMENT #141=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Tm: 80.8 C TaOpt: 58.7 C GC: 54.9" 1) COMMENT #142=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #143=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #144=(COligo "" "CCCATGTTCTTTGCCCACACA" COMMENT "Sense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " CCCATGTTCTTTGCCCACACA" 1)) COMMENT #145=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #146=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 21 Tm: 57.9 C GC: 52.4" 1) COMMENT #147=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -158.7 kcal/mol dS: -403.4 cal/mol dG: -36.6 kcal/mol" COMMENT 1) COMMENT #148=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #149=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #150=(COligo "" "AAAGGCTCTGGGAGGGTCTAGG" COMMENT "Antisense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " AAAGGCTCTGGGAGGGTCTAGG" 1)) COMMENT #151=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #152=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.7 C GC: 59.1" 1) COMMENT #153=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -177.2 kcal/mol dS: -457.5 cal/mol dG: -39.0 kcal/mol" COMMENT 1) COMMENT #154=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 1.1" 1) COMMENT #155=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 6.7" 1))) COMMENT "#2: Product of length 426 (rating: 171)")) COMMENT #156=(CPCRFolderPar COMMENT (CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 0 COMMENT (20 COMMENT #157=(CPCRData (CStringList) (CStringList) COMMENT "atgttctttgcccacacagcc" 0 COMMENT "aaaggctctgggagggtctagg" 0 1373 1795 423 0 COMMENT 0) 0) 1 2 1 0 182) COMMENT (CObjectList COMMENT #158=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Contains region of the molecule from 1373 to 1795" COMMENT 1) COMMENT #159=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Tm: 80.7 C TaOpt: 58.5 C GC: 54.6" 1) COMMENT #160=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #161=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #162=(COligo "" "ATGTTCTTTGCCCACACAGCC" COMMENT "Sense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " ATGTTCTTTGCCCACACAGCC" 1)) COMMENT #163=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #164=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 21 Tm: 56.3 C GC: 52.4" 1) COMMENT #165=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -160.8 kcal/mol dS: -411.4 cal/mol dG: -36.4 kcal/mol" COMMENT 1) COMMENT #166=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #167=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #168=(COligo "" "AAAGGCTCTGGGAGGGTCTAGG" COMMENT "Antisense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " AAAGGCTCTGGGAGGGTCTAGG" 1)) COMMENT #169=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #170=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.7 C GC: 59.1" 1) COMMENT #171=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -177.2 kcal/mol dS: -457.5 cal/mol dG: -39.0 kcal/mol" COMMENT 1) COMMENT #172=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 0.4" 1) COMMENT #173=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 6.7" 1))) COMMENT "#3: Product of length 423 (rating: 171)")) COMMENT #174=(CPCRFolderPar COMMENT (CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 0 COMMENT (20 COMMENT #175=(CPCRData (CStringList) (CStringList) COMMENT "gttctttgcccacacagccc" 0 COMMENT "aaaggctctgggagggtctagg" 0 1375 1795 421 0 COMMENT 0) 0) 1 2 1 0 182) COMMENT (CObjectList COMMENT #176=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Contains region of the molecule from 1375 to 1795" COMMENT 1) COMMENT #177=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Tm: 80.8 C TaOpt: 58.7 C GC: 54.9" 1) COMMENT #178=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Sense Primer: " 1) COMMENT #179=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #180=(COligo "" "GTTCTTTGCCCACACAGCCC" COMMENT "Sense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " GTTCTTTGCCCACACAGCCC" 1)) COMMENT #181=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #182=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 20 Tm: 56.7 C GC: 60.0" 1) COMMENT #183=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -157.4 kcal/mol dS: -401.2 cal/mol dG: -36.0 kcal/mol" COMMENT 1) COMMENT #184=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Antisense Primer: " 1) COMMENT #185=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #186=(COligo "" "AAAGGCTCTGGGAGGGTCTAGG" COMMENT "Antisense primer for PCR amplification of a region of Ple219 (pEMS1182)" COMMENT 0 (CStringList) 0) 1) 1 2 2 0 181) COMMENT " AAAGGCTCTGGGAGGGTCTAGG" 1)) COMMENT #187=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Similarity: 100.0%" 1) COMMENT #188=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Length: 22 Tm: 56.7 C GC: 59.1" 1) COMMENT #189=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " dH: -177.2 kcal/mol dS: -457.5 cal/mol dG: -39.0 kcal/mol" COMMENT 1) COMMENT #190=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Tm Difference: 0.0" 1) COMMENT #191=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "GC Difference: 0.9" 1))) COMMENT "#4: Product of length 421 (rating: 171)")))) COMMENT "PCR Analysis")))) COMMENT (CGraphView COMMENT (CWStyleSheet COMMENT (CObjectList COMMENT #192=(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 #193=(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 #194=(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 #195=(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 #196=(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 #197=(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 #198=(CWidgetStyle "Axis" 1 (LOGPEN 0 0 10079436) 2 1 COMMENT (LOGBRUSH 0 13434879 0) 0 0 1 (LOGSHAPE 10 1 0 0 0)) COMMENT #199=(CWidgetStyle "Line 2" 1 (LOGPEN 0 0 6723840) 8 0 0 0 1 COMMENT (LOGSHAPE 1 1.9 0 0 0)) COMMENT #200=(CWidgetStyle "RSite" 1 (LOGPEN 0 0 10053171) 8 0 0 0 1 COMMENT (LOGSHAPE 1 1.9 0 0 0)) COMMENT #201=(CWidgetStyle "Short Signal" 1 (LOGPEN 0 0 13395507) 10 0 0 0 1 COMMENT (LOGSHAPE 1 1.9 0 0 0)) COMMENT #202=(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 #203=(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 #204=(CWidgetStyle "Mark 1" 0 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 2 7 48 2 2 "Windings") 0.7 0 1 2 "?" COMMENT 0) COMMENT #205=(CWidgetStyle "Motif Label" 1 (LOGPEN 0 0 16744512) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Arial") 0.611111 8388608 COMMENT 1 65535 "@N (@H)" 0) COMMENT #206=(CWidgetStyle "Fragment Label 2" 1 (LOGPEN 0 0 0) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 49 "Courier New") 1.05 0 1 48 COMMENT "@F bp (molecule @L bp)" 0) COMMENT #207=(CWidgetStyle "Fragment Label 1" 1 (LOGPEN 0 0 0) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Arial") 0.91 0 1 1 COMMENT "Fragment of @N" 0) COMMENT #208=(CWidgetStyle "Shape 4" 1 (LOGPEN 0 0 0) 1 1 COMMENT (LOGBRUSH 2 8388608 5) 0 0 0) COMMENT #209=(CWidgetStyle "Shape 2" 1 (LOGPEN 0 0 0) 1 1 (LOGBRUSH 0 128 0) 0 COMMENT 0 0) COMMENT #210=(CWidgetStyle "Shape 0" 1 (LOGPEN 0 0 0) 1 1 (LOGBRUSH 0 0 0) 0 0 COMMENT 0) COMMENT #211=(CWidgetStyle "ORF" 1 (LOGPEN 0 0 16384) 8 0 0 0 1 COMMENT (LOGSHAPE 7 0.2 3.41182 2.86186 0.609808)) COMMENT #212=(CWidgetStyle "Line 4" 1 (LOGPEN 0 0 32768) 8 0 0 0 0) COMMENT #213=(CWidgetStyle "Line 3" 1 (LOGPEN 0 0 16711680) 8 0 0 0 0) COMMENT #214=(CWidgetStyle "Line 1" 1 (LOGPEN 0 0 16711680) 1 0 0 0 0) COMMENT #215=(CWidgetStyle "Short Promoter" 1 (LOGPEN 0 0 128) 6 0 0 0 0) COMMENT #216=(CWidgetStyle "Motif" 1 (LOGPEN 0 0 0) 1 0 0 0 0) COMMENT #217=(CWidgetStyle "Line 0" 1 (LOGPEN 0 0 0) 8 0 0 0 0) COMMENT #218=(CWidgetStyle "Void" 0 0 0 0 0) COMMENT #219=(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 #220=(CWidgetStyle "Position" 1 (LOGPEN 0 0 0) 1 0 0 0 0) COMMENT #221=(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 #222=(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 #223=(CWidgetStyle "Range" 1 (LOGPEN 0 0 0) 1 1 COMMENT (LOGBRUSH 0 16777215 0) 0 0 0) COMMENT #224=(CWidgetStyle "Range 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 #225=(CWidgetStyle "ORF Label" 1 (LOGPEN 0 0 49216) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Times New Roman") COMMENT 0.611111 0 1 65535 "@N" 0) COMMENT #226=(CWidgetStyle "CDS Label" 1 (LOGPEN 0 0 4227264) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Arial") 0.555556 255 1 1 COMMENT "@N" 0) COMMENT #227=(CWidgetStyle "Shape 5" 1 (LOGPEN 0 0 0) 3 1 COMMENT (LOGBRUSH 0 16777113 0) 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 7 48 2 50 "Arial") 0.9 0 0 1 COMMENT (LOGSHAPE 9 1 0.8 1.8 0)) COMMENT #228=(CWidgetStyle "CDS" 1 (LOGPEN 0 0 0) 1 1 (LOGBRUSH 2 39423 3) 0 0 COMMENT 1 (LOGSHAPE 9 1 0.8 1.8 0)) COMMENT #229=(CWidgetStyle "Label 2" 1 (LOGPEN 0 0 4227264) 1 0 1 COMMENT (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Arial") 0.944444 8388608 COMMENT 1 1 "@N" 0) COMMENT #230=(CWidgetStyle "Label 3" 1 (LOGPEN 0 0 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 #231=(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 -16869) 1 1 0 1 1 COMMENT (mapper: 18.2682 -18.0184 40.0378 40.0378 0.01 10 -1 18291 18291 1 0 0) COMMENT #232=(CGroupWidget (CWidget 0 (0 0) 1 2 0 0 Nil -1367 100) COMMENT (CObjectList COMMENT #233=(CGroupWidget (CWidget 1 (0 0) 1 2 0 0 Nil -953 100) COMMENT (CObjectList COMMENT #234=(CAxis COMMENT (CWideLine COMMENT (CWidget 0 (0 0) 1 2 0 0 #198# 47956620 0) COMMENT (LOGPEN 0 0 10079436) 2 (LOGBRUSH 0 13434879 0) 1 COMMENT 6.27453 6.27253 1 0.0214037) 0.0342966) COMMENT #235=(CLabel COMMENT (CWidget 1001 (0 0) 1 2 0 0 #195# 4063824 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 58 22 0 0 400 0 0 0 0 3 2 1 34 "Verdana") COMMENT 1.74233 0.833333 0 "Ple219 (pEMS1182)" "@N" 1 0 COMMENT 0.871165 0 -10 11.4392 1.47361 Nil) COMMENT #236=(CLabel (CWidget 1002 (0 0) 1 2 0 0 #194# 543 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 38 14 0 0 400 0 0 0 0 3 2 1 18 "Georgia") COMMENT 1.74233 0.555556 0 "18291 bp" "@L bp" 16 0 -0.871165 COMMENT 0 -10.8 3.12205 0.949103 Nil)) (CObjectList)) COMMENT #237=(CGroupWidget (CWidget 10 (6 0) 1 2 0 0 Nil -1232 100) COMMENT (CObjectList COMMENT #238=(CGroupWidget COMMENT (CWidget 21 (7 21 0) 1 2 0 0 Nil -256 100) COMMENT (CObjectList COMMENT #239=(CScratch COMMENT (CWidget 0 (3 #5# 0) 1 2 0 0 #201# 629 100) COMMENT (LOGPEN 0 8 13395507) 10 1 5.59789 1.9 COMMENT 0.082322 1) COMMENT #240=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 24492700 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 COMMENT "FRT (F5 mutant)" "@N" 1 13.4289 6.96932 0 COMMENT -10 7.41799 1.17389 #239#) COMMENT #241=(CScratch COMMENT (CWidget 0 (3 #6# 0) 1 2 0 0 #201# 641 100) COMMENT (LOGPEN 0 8 13395507) 10 1 5.32454 1.9 COMMENT 0.082322 1) COMMENT #242=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 24494596 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 "NLS" "@N" 1 COMMENT 12.873 0 0 -10 1.99811 1.17389 #241#) COMMENT #243=(CScratch COMMENT (CWidget 0 (3 #7# 0) 1 2 0 0 #201# 3670472 COMMENT 100) (LOGPEN 0 8 13395507) 10 1 5.29779 1.9 COMMENT 0.082322 1) COMMENT #244=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# -1437 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 "FRT" "@N" 1 COMMENT 12.7446 -1.74233 0 -10 1.92318 1.17389 #243#) COMMENT #245=(CScratch COMMENT (CWidget 0 (3 #8# 0) 1 2 0 0 #201# 3670472 COMMENT 100) (LOGPEN 0 8 13395507) 10 1 5.23366 1.9 COMMENT 0.082322 1) COMMENT #246=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 1944545584 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 COMMENT "SV40 early polyA signal" "@N" 1 16.4073 COMMENT -3.48466 0 -10 11.0645 1.17389 #245#) COMMENT #247=(CWideArrow COMMENT (CWideLine COMMENT (CWidget 0 (3 #9# 0) 1 2 0 0 #197# 0 100) COMMENT (LOGPEN 0 0 6723840) 1 COMMENT (LOGBRUSH 0 10079334 0) 0.835356 5.33689 COMMENT 5.58382 1 0.082322) 0.8 1.8 0) COMMENT #248=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 23506756 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 COMMENT "EGFP (mutated stop TAA -> TTA)" "@N" 1 COMMENT 18.3413 3.48466 0 -10 15.0358 1.17389 #247#) COMMENT #249=(CScratch COMMENT (CWidget 0 (3 #10# 0) 1 2 0 0 #201# 3670472 COMMENT 100) (LOGPEN 0 8 13395507) 10 1 5.58622 1.9 COMMENT 0.082322 1) COMMENT #250=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 3670592 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 COMMENT "Kozak consensus" "@N" 1 14.5547 5.22699 0 COMMENT -10 7.99245 1.17389 #249#) COMMENT #251=(CScratch COMMENT (CWidget 0 (3 #11# 0) 1 2 0 0 #201# 0 100) COMMENT (LOGPEN 0 8 13395507) 10 1 5.33689 1.9 COMMENT 0.082322 1) COMMENT #252=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# -879 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 COMMENT "Tetracysteine residue" "@N" 1 16.2902 COMMENT 1.74233 0 -10 10.0405 1.17389 #251#) COMMENT #253=(CScratch COMMENT (CWidget 0 (3 #12# 0) 1 2 0 0 #201# 641 100) COMMENT (LOGPEN 0 8 13395507) 10 1 2.11198 1.9 COMMENT 0.082322 1) COMMENT #254=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# -740 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 "I-Sce I" "@N" 1 COMMENT -11.9333 -6.96932 0 -10 3.02214 1.17389 #253#) COMMENT #255=(CWideArrow COMMENT (CWideLine COMMENT (CWidget 0 (3 #13# 0) 1 2 0 0 #197# 0 100) COMMENT (LOGPEN 0 0 6723840) 1 COMMENT (LOGBRUSH 0 10079334 0) 0.835356 5.60063 COMMENT 6.21865 1 0.082322) 0.8 1.8 0) COMMENT #256=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 393244 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 "TAC3-B" "@N" 1 COMMENT 6.87217 13.9386 0 -10 3.47172 1.17389 #255#) COMMENT #257=(CWideArrow COMMENT (CWideLine COMMENT (CWidget 0 (3 #14# 0) 1 2 0 0 #197# 0 100) COMMENT (LOGPEN 0 0 6723840) 1 COMMENT (LOGBRUSH 0 10079334 0) 0.670712 5.60063 COMMENT 5.73096 1 0.082322) 0.8 1.8 0) COMMENT #258=(CLabel (CWidget 0 (0 0) 1 2 0 0 #193# 0 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 "Prom region" COMMENT "@N" 1 11.2891 8.71165 0 -10 5.86945 1.17389 COMMENT #257#) COMMENT #259=(CWideArrow COMMENT (CWideLine COMMENT (CWidget 0 (3 #17# 0) 1 2 0 0 #197# 870 100) COMMENT (LOGPEN 0 0 6723840) 1 COMMENT (LOGBRUSH 0 10079334 0) 1 5.73096 6.21865 1 COMMENT 0.082322) 0.8 1.8 0) COMMENT #260=(CLabel COMMENT (CWidget 0 (0 0) 1 2 0 0 #193# 178 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 COMMENT "Regions 1, 2, 3" "@N" 1 7.65913 15.681 0 -10 COMMENT 6.84353 1.17389 #259#)) (CObjectList)) COMMENT #261=(CGroupWidget COMMENT (CWidget 22 (7 22 0) 1 2 0 0 Nil -325 100) COMMENT (CObjectList COMMENT #262=(CWideArrow COMMENT (CWideLine COMMENT (CWidget 0 (3 #1# 0) 1 2 0 0 #209# 0 100) COMMENT (LOGPEN 0 0 0) 1 (LOGBRUSH 0 128 0) 1 COMMENT 0.00240076 0.345709 1 0.082322) 0.8 1.8 0) COMMENT #263=(CLabel (CWidget 0 (0 0) 1 2 0 0 #193# 0 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 46 17 0 0 700 0 0 0 0 3 2 1 34 COMMENT "Arial") 1.74233 0.666667 0 "AmpR" "@N" 1 COMMENT -4.15533 12.1963 0 -10 2.87229 1.17389 #262#) COMMENT #264=(CWideArrow COMMENT (CWideLine COMMENT (CWidget 0 (3 #2# 0) 1 2 0 0 #209# 0 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COMMENT #279=(CGroupWidget (CWidget 11 (0 0) 1 2 0 0 Nil -116 100) COMMENT (CObjectList) (CObjectList)) COMMENT #280=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -395 100) COMMENT (CObjectList) (CObjectList))) (CObjectList))) COMMENT (CSeqView 10 10 (CObjectList) (CObList) 1 COMMENT (CObList #281=(AnlzFontBackColorItem 1581 1665 40304639) COMMENT #282=(AnlzFontBackColorItem 1795 1960 40304639) COMMENT #283=(AnlzFontBackColorItem 1773 1794 43633766) COMMENT #284=(AnlzFontBackColorItem 1689 1772 40304639) COMMENT #285=(AnlzFontBackColorItem 1666 1688 43633766) COMMENT #286=(AnlzFontBackColorItem 159 1580 50331392))) (CObList) 1919252069 COMMENT (CStringList) 1114668146 1914717249 (CObList))) FEATURES Location/Qualifiers misc_recomb 12153..15964 /vntifkey="86" /label=5'\junction /note="The 5 HPRT probe anneals to genomic sequence ~560bp upstream of the BamHI site at 9263" misc_marker 17284..18284 /vntifkey="22" /label=AmpR misc_marker 10681..12127 /vntifkey="22" /label=AmpR misc_recomb 7795..10685 /vntifkey="86" /label=HPRT\3'\homology /note="Consists of part of Intron 2, all of Exon 3, and part of Intron 3" misc_recomb 3088..7794 /vntifkey="86" /label=HPRT\complementary\sequence /note="Consists of human HPRT promoter, human Exon1, Intron1, Exon2, and part of Intron2" misc_feature 1970..2003 /vntifkey="21" /label=FRT\(F5\mutant) misc_feature 2767..2839 /vntifkey="21" /label=NLS misc_feature 2845..2878 /vntifkey="21" /label=FRT misc_feature 3032..3082 /vntifkey="21" /label=SV40\early\polyA\signal misc_feature 2011..2730 /vntifkey="21" /label=EGFP\(mutated\stop\TAA\->\TTA) misc_feature 2004..2014 /vntifkey="21" /label=Kozak\consensus misc_feature 2731..2766 /vntifkey="21" /label=Tetracysteine\residue misc_feature 12134..12151 /vntifkey="21" /label=I-Sce\I misc_feature 160..1961 /vntifkey="21" /label=TAC3-B misc_feature 1582..1961 /vntifkey="21" /label=Prom\region primer 1155..1178 /vntifkey="27" /label=oEMS2646 /note="56.6" primer 1667..1689 /vntifkey="27" /label=oEMS2622 misc_feature 160..1581 /vntifkey="21" /label=Regions\1,\2,\3 BASE COUNT 4782 a 4206 c 4179 g 5123 t 1 others ORIGIN 1 cggccgcacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc 61 acgaggccct ttcgtcttca agaattaatt ccgatcatat tcaataaccc ttaatataac 121 ttcgtataat gtatgcgaat tcgagctcct aggccggccg agccagctag tgaggtaata 181 agaaaggaaa agaaggaaaa ttgtctccgg gctccttgac cggctgagct ctgggggggt 241 gtttagagag actgcggtgg gtggaggggc tgcgggggga gttaaggatg gggctcaggt 301 cgcaggtggc cagtggactg attcattaag tgtgtccctg gaggaaagaa gtgagcatcc 361 ctgtcttggc agaaactggg gtcctttggc gatttagcct gagctttgag ctgagataga 421 acctggggaa tcatctgttt tcaggcgggt gaggggctag agcctgcctt gtttgggagg 481 agggtggctc tgttcagaat aggggtagct caggctctgg ccagccttct cccgccccca 541 acagctcccc ccatccttga cttctcagaa tcaggccgag aagagcctat ctggccgaga 601 gtggggtggt gacctgcgcc tcatcgcccc cgctctccat ctcatctcct gctcccaggg 661 cccaaattgt cgtcactttc ccagtgaagt gtctggtcat tttcagaagc aatttcagga 721 gaacatgcag ctgccgctcc ctatcctgca tttcccttca cagggctgaa ggcactgtca 781 gctccctggg ctgggggtga tgggagaggg gaagggctag ggccctcacc cctgtcctca 841 ctgtgcaaca tagcctagca ttagctcttt caagtctttg ctaatcccag agatcaaggg 901 gtgatcaact ctccctgcca tccccttgtt ccccgcaccc cccgccccgg ctcccccacc 961 atccttggct cctgccatcc tctttgagat gctgcatcat caaaggacat tatttatggt 1021 gtacctttgc tgaagccctg cttccctggt gccagggctt gggagcaggg atgggtgggt 1081 tggtggggga gaggggtgga tgcagagatt ggacccagga ggcttttagt cctcagctct 1141 tggcttaaca cctcctcctc ttacacaccc aactccctcc agcctgccca gcttgggcct 1201 tcagctccag attggtgggg ttaggagagg aggaggaggg agatggatgg aaccaattag 1261 gaacagcacc tgggctcctc acaggaatga accagtcatg ccatttgcat gtaaacagct 1321 tcccacttct ctcctcatcc taccaaatgc tcccaaccct gggttctggc ccatgttctt 1381 tgcccacaca gccctgtaat tagctgggta atgagaagct tttaatgagt cccattagca 1441 tctcgtgtaa taaagaggcc ttgagaccca gctgctgtcc tcactttggg atgaacacgg 1501 gtccctgtgt agccagtgac ttctgtcagt acagtctaag ttctcggatg gggtgggaga 1561 caaacatttc aggaccccag ctcaaactgt acacaaacca tacacagcag ccagctcggg 1621 tgctgttagg aaatggtctc actgctgggt ctgtggggta tgtgtgtgtc tgggtgtgtg 1681 gctactgtct gcatcctcct cccccctaca gcctccccgc ctcccctcca gccaccctgg 1741 gattggtgac tctcagcccc tcccctcagc tcccctagac cctcccagag cctttatcag 1801 ggagctggga ctgagtgact gcagccttcc tagatcccct ccactcggtt tctctctttg 1861 caggagcacc ggcagcacca gtgtgtgagg ggagcaggca gcggtcctag ccagttcctt 1921 gatcctgcca gaccacccag cccccggcac agagctgctc cggcgcgccg aagttcctat 1981 tcttcaaaag gtataggaac ttccgccacc atggtgagca agggcgagga gctgttcacc 2041 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg 2101 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 2161 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag 2221 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc 2281 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc 2341 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac 2401 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac 2461 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac 2521 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc 2581 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa 2641 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc 2701 actctcggca tggacgagct gtacaagtta tttctgaact gctgcccggg ctgctgcatg 2761 gaaccggatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtagatcca 2821 aaaaagaaga gaaaggtaat ataagaagtt cctattctct agaaagtata ggaacttcag 2881 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta 2941 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 3001 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 3061 aataaagcat ttttttcact gcgaattctc gtaaaactct tcatggcagt agttattatt 3121 ctctctctct ctctttttct tttttcttga gacaggatat ttctctgttg cccaggctgg 3181 agtgcagtgg cacagtcttg gctcactgca gcctggacct cctgggctca agccatcctc 3241 ccacctcagc ctcccaagta gctggggcta caggcacatg gccaccaggc cagataattt 3301 ttcatttttg tagagactga gtctcaccat gttacccagg tttattattc tcatttttta 3361 gatgaagaga ctgaggtcca gagaagctca atgacttgcc tagttttaca aatctcctgc 3421 catcacatac ccctcagcgt ccttaataag agggaggcca ccaactatgt gctgggcact 3481 gtggtggatg ctggagctat agggttgagt atataagaaa tggtgttgct ggagcaactg 3541 ttgcttgctt acctgaccta tctgagaatt aattagcagg ggaacatatt tttgttttca 3601 gattcaatat aagaacttgt gtgggcaaaa ataaagatca gtagtaataa cagtagttcc 3661 catttgctga ctgtactgtc ctaagtgcat atatatatac atacacacac gcatacctat 3721 actcctctaa tactcaaaat gatcctgttt atgtattgtt aatatgctca ttttattttt 3781 aaatttttat ttatttttat ttttatttat ttttgagacg gagtctcatt ctgtcgcgga 3841 ggctgaagtg cagtggtgcg atctcagctc agtgcgacct ccgcctcccg ggttcaagtg 3901 attctcctgc ctcagctccg gactagctgg gattacaggc gcccgcctcc acgcccagct 3961 aatttttgta tttttagtag agatggggtt tcgccatgtt ggccaggctg gtctcgtact 4021 cctgaccttg agtgatccac ctgcctcggc ctcccaaagt gctgggatta caggcatgag 4081 ccaccgcgcc gggctaatat gctcatttta gtgaggcaaa aatagaggct cagagtctga 4141 tttgtacaaa actacagagc agttaagtgt cctctcagat gtgtaccctg atctgggtga 4201 ctctaggact ctaggtctca actgttacaa ccagttaagg gtttggggaa gcactgggcc 4261 aagagtcagg aaaatggaag ccacaggtag tgcaaggtct tgggaatggg acgtctggtc 4321 caaggattca cgcgatgact ggaacccgaa gagccggggc ccggtttacg gccgccatga 4381 agcaacgcgc gccggtaggt ttgggaatca gggagccctc tgaataggag actgagttgg 4441 gagggaaagg ggcttcgctg ggggagcctc ggcttcttct gggagaaaat tcccacggct 4501 acctagtgag cctgcaaact ggtaggcgcc ggcgtaggcg cgcgggcggg gccgggggcg 4561 gggcctgcgg ggcgtggcgg ggcgggcaga gggcggggcc tgcttctcct cagcttcagg 4621 cggctgcgac gagccctcag gcgaacctct cggctttccc gcgcggcgcc gcctcttgct 4681 gcgcctccgc ctcctcctct gctccgccac cggcttcctc ctcctgagca gtcagcccgc 4741 gcgccggccg gctccgttat ggcgacccgc agccctggcg tcgtggtgag cagctcggcc 4801 tgccggccct ggccggttca ggcccacgcg gcaggtggcg gccgggccct gaggcgcggg 4861 atcgatccat gacacccccc cgccccccat atctcactat aattcactgt caaatggctt 4921 ttatgatttg gagtttgggg atacttttaa agacagggtc ttggtagctc ctagatggct 4981 tgggattcac ttagtaaccc ttgctggcct aaaattcagg aacttctacc tcatcttccc 5041 aagtactggg ttagtaggga tgtccagcat atttagcttg ctttaatgat tttgctgtac 5101 ttttttctgt gtgtacaggt gcatgtatat ttataaaact aacataatat attttacccc 5161 caaagtacca taaccctaat taatgctatt aaatgttttg tatgctgggc tctgggcata 5221 ctaatataac caatattata caatgcaaaa caattttgga atatcttctg taaaattatc 5281 ttattagtaa gttgtaacca tctcagactt ctaagtttga gtttgtaaag gatcagtttt 5341 ttttggtgac ctacacacac acacacacac acacacacac acacacacac aggggggtgg 5401 attctgagaa tttagagaat ttagatggtt aataccagtt tttattttta ttctgtgtga 5461 tatttattta ttaagattta tttatttatt acatgtaggt acactgtatc tgtcttcaga 5521 cacaccagaa gagggcatca gatctcgtta cagatggttg tgagtcacca tgtggttgct 5581 gggatttgaa ctcaggacct ctggaagaac agtcagtgct cttaaccgtt gagccatctc 5641 accagccccg tgatatttta tttaacttac tagaatatac tgtgttaaaa catattttag 5701 gggattgggg agatggccca ttctgtaaag tgtctgccat gtcatcttca ggacctgcgt 5761 ttgtaacccc agcatggaga agacagagac agggagaccc ctgggcttcc tggcaagcta 5821 gactagcaaa tttggttaac taggttcaat gagagactca gtctcaaaac ataagaagtg 5881 aattttttag acccaatttc aatctctgaa ctccatacgt atgcacatgt acctgcatat 5941 aaatatatat acacacccat aaacacatgc acacacaaaa aatggttcaa aagttcttaa 6001 aatgtataag agtagaaagt gtagagccgg gagtggtggc gcacgccttt aatcccagca 6061 cttgggagca gaggcaggtg gatttctgag ttcgaggcca gcctggtcta cagagtgagt 6121 tccaggacag ccagagctat acagagaaat cctgcctcaa aaaaccaaaa ataaagtgta 6181 tatactaatc tttcatgcct aaaatattta tttttgccac atttttcata cctctttaat 6241 atttttcttt ttctctttgt gcttcaaagg aaattttgtg tcatgtttta cttttcattc 6301 ttgactctat tcagcagtaa gacgcagcat ggctttctta gagaagtata atctattaca 6361 aaacggtaac atttaactgc tctacatacc ctcttgtagc taaattcctc tgttaaacta 6421 agacttatta tctaaacgta attaaatgtt gatatcttat aatgctccac tttgaaacag 6481 ctgtctttct tttgcagatt agcgatgatg aaccaggtta tgacctagat ttgttttgta 6541 tacctaatca ttatgccgag gatttggaaa aagtgtttat tcctcatgga ctgattatgg 6601 acaggttagt aagacctcga ttgaagttat ttttagaggg tgtgtgtgtg gtgtgtgtgt 6661 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt aagagagaga ggagaggcat gtgtgatata 6721 tatgtgaagt gtattgcatg tacatggaat ccagagaagg acattaggtg tcctactctg 6781 ttgctctctt attcccttga atcagtgtct cttgctgaac ctggggcata catactgatt 6841 tggctaggct gcctggctag caagacccag ggatctgctt gtctctgccc cccagtgctg 6901 acgttacaag caggtgcaac cacatctggt ttttatatgg gtactggggg atctgaactt 6961 agggcctgat gcgtgcccaa gtgctctcac ccgccaagcc acctctccag ccaaagggtt 7021 ttttttcttg tattaattct aacttagcaa ctgttgagta gttgatgttt gaatataagc 7081 taagccaact tattagaata aatgttttct attttttact tgtatgtgtg agtgcctgca 7141 tgtatgtatg tgaagcatat gtattccagg atgtcagaat tgtgatggat tccctggaac 7201 tggagttaca gaccattgtg aaccactgtg tatgggtgct gggaactgaa ctttaggtcc 7261 tctgcaaaaa cagtaaggat cttaactgat aaactatctt tctagcccta aggctttcta 7321 cttttaaagt tcgtttctct ttctctctct ctctctctct ctctctctct ctctccctct 7381 ccctccctcc tcctcctcct cctctttccc ccctctcgcc tcctctttct ctctcccctc 7441 cctctctctc tctcacacac acacacagac agagagagga gagagaaagg aagccttact 7501 aattaattac ctagttttag acataactag tgggaatcca tagtaagtaa atcaacattc 7561 ttgaagagag gttagaagaa ttggctctcg ttgtccatcg aggctgtata agattattct 7621 tctgttttct cagcttacca tacatatttt ctacgtagac aataccaatt caaatacact 7681 gtacgtaaag tatatgcgca cacacacaca cacacacaca cacacacaca cacaagttac 7741 atattgtcat atagtactaa aatttatgga ctttttcata ttaatacatc tagagctatc 7801 atgtttttta tcttttaaaa attactttac ggggctacag agatgtctca gcagttagga 7861 gcactgactg ctcttccaga ggtcctgagt tcaattccca gcaaccacat ggtggctcac 7921 aaccatctgt aatgggatct gatccactct tatggtgtgt ctgaagacag tgacagtgta 7981 ctcatataca tgaaataaat ctttaaaaaa ttactttacc atcagggtat atatatatat 8041 atatatgtgt gtgtgtgtgt gtgtgtgtat gaagtatttg ctatttgaga gggtctcact 8101 gtttagctca gtctggcttg gaacgttctg tatagcccag actggcctca actcagcagt 8161 ccccctgaaa ggctctcata tgtggagatt acaggtatgt gctactgtgc ccagccctac 8221 atgttacttt gtttttgaaa tagggtatca aatagcttag actggcttct aagtcagtgt 8281 gtagctaagg atggtctgag cttctgaacc tgctacttca actcctggtg tgccatcatg 8341 gccagtttgc acagttgtga gagtagaact caggacctcc tatgtgttac tagacatgtt 8401 ctctaccagc tgagctacat cccaattcgc ctacatttac ttttattttt taaaagattt 8461 atttatgtat acagcactct gcctgcatgt acatctgcat gccagaagag ggcaccagat 8521 ctcatcatac atagttgtga gccaccatgt ggttgctggg gattgaactc agggcctctg 8581 gaagagcagt cagtgctctt aacctctgag ccatctctgt agccccctac atttatttta 8641 aagattatag gctattctat tcatagatgt gccataactg atttaaccag atcttcatta 8701 ttttgtactt agtagagtat attagaggga atgaatctta aatacacatt ttgacacttg 8761 ctcagcttta gacaagtaaa aatcactggt caaggaaatg gtgcttagtg gatgcttctg 8821 aactgtcaaa ttgtcctcca acaagttaca tcattttata cttccaacgt tatgtgagag 8881 ttcatttttc tttaggctct tctagtattt tatatttgag aaataaaatt taaccaaaat 8941 ttgccaatct agtaagcttg tttttatttg tatctttaaa ttttatgata gatgttatag 9001 tgtactctcc tctcccctcc cctcccctct cctcctcagc aggatcttgt tatgtagccc 9061 taagggatct aatacttact ctgtagtcca ggctgtcctt cagctcatga gacacaccat 9121 catgcttggc atgtttcttg ttctgaaagg tatctggttt taattttggt tgagctggtg 9181 tgttggtgta tatctgtaat ctcagcacag gcaagtagag acaaggatta tgattacagt 9241 ccagcctggg ctatttagag ggattctctt taatctttcc tcatgcccca aaatcttacc 9301 tttggtatat gaaaaatagt ctccacttct gcaaaatatt gctttatgaa gtaagaatta 9361 attcccttca tagagacaag gaatgtgtcc tgtaaaagtt taatgtgtaa gaagtatttg 9421 ttataaaaga taaatattca gaatcttctt tttaattcct gattttattt ctataggact 9481 gaaagacttg ctcgagatgt catgaaggag atgggaggcc atcacattgt ggccctctgt 9541 gtgctcaagg ggggctataa gttctttgct gacctgctgg attacattaa agcactgaat 9601 agaaatagtg atagatccat tcctatgact gtagatttta tcagactgaa gagctactgt 9661 gtaagtataa ttaacttata attaaaaaaa tagggccatt ctagttttat ctatattttt 9721 tttaaacttg tgcaaactat gctacataat caacttactt gattatgtaa tatatatagt 9781 aatagttgga attactatta ttacatattt atttttctgt gctggggtag aacccaggga 9841 actaagtact gtaataaccg agcttcattc atcctagtca tttttaattt tttttttttt 9901 ttttgttttt tcgagacaag gtttctctgt gtagccctgg ctgtcctgga actcactctg 9961 tagaccaggc tggcctcgaa ctcagaaatc tgcctgcctc tgcctcccag tgctgagatt 10021 aatgacgtgc accaccactg cctggcattt ttttaaagtt ttaatttgag accctttctc 10081 actaacttgc tcacactatc ctcgaactca ttacgtgggc tggtctagaa atttcagtca 10141 acttgcctca gcttctcaag tagctgggat tacaggcagg tgctaccact tctggccatg 10201 atcattgttg ttgttgctat tattattatt attatttcac tgatacttca tatcacaaca 10261 cagaatagta actgtagttc atgaccaaca aaaaagttgt acagtttctc tgttgtctta 10321 ttttgatctg gattttgatt tgtttcatgg cactgggcag gctaatctca aactgcatgg 10381 ctcaaatagt ccttctgtct taatctccca tgttctggaa gtataggtat cactacacct 10441 agctgtattt tcatatttta aaaaaaacgc ctaagaattg ctattgaatg aggcggagag 10501 atggctcagc agtcacaagt acttgtagag gactggggtt tggttactag cacctcaaag 10561 atggcttaca attatctgta gctccaattc cagaggatct gtgtggtaaa gctgcatgta 10621 tatgggcatg caggtaaaac tgctaaacat ataaaaatag ctcagtctta agagaattgg 10681 tcgacgtaag ctcattcgag tcgacttttt cgccctttga cgttggagtc cacgttcttt 10741 aatagtgdga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 10801 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 10861 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg 10921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 10981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 11041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 11101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 11161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 11221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 11281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 11341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 11401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 11461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 11521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 11581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 11641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 11701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 11761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 11821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 11881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 11941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 12001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 12061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 12121 ctaccaggtc gagtagggat aacagggtaa tggatcctcg ttgggcacct gcactcatgt 12181 acacaggccc acatatagat gtacacacgc atatcattca aaatagaaat tatttataca 12241 agtagtgcgt tggagagctg gctcaggagt tgttaatgct tgctaccctt gcaaaggacc 12301 caagttcaat ttccagcacc tgtgttgggc atctcacaac cacatgtaaa tctagtttca 12361 ggggatttca cccctcttct gaactcttca ggcacttata cccacaaatg gacatctcca 12421 catagagaca catactttta aaaaagtaat ctagggtcta gagaaatggt tatagaccac 12481 ggactgggat cagttcccag aagccacatc ctgtggctca caattgcctg tgaatcaagt 12541 tctagatctg gtggcctcgt ctagcctgca gtcatcaaca aacacatgca cacatccata 12601 cacacataat ttaaaagaaa gcgaaagtaa cctataatgc taaacaaagc acattctcag 12661 tgtagactgc agtggtgatg aaccttgaaa atatattaat agaagccaga cacgtaagtc 12721 acaaatggta taactcaatt tatatgaaat tcccagagag tccaaagtta aaaaaaacaa 12781 aaaggcagat tattggttgt cagtagctac agagatggga cattgaagat attaataggt 12841 atagggttga ggattcatgg cctttccctc ttccatgtta gcatgtctat tgttgttaac 12901 cttattcaac tcatgttagg gcaatcatat tgatgagact taatgagtat agcttgacat 12961 tcctagaaga cagagtctca catcagactc tctgatcctc tgactcttag aatctttcaa 13021 caccctcttc tgaaatgatc tctgagcctt tggttgtgac tgaattatga gaaagaattt 13081 tatgttgaat gaattttact tcagcatttt tacaaaactg aagctgggga tggcgtttca 13141 tgcgtgcagt ccctaccatt tgcctataag cctggaggag gcagaagtag aattaggctt 13201 ttcagggaca tcttcagctg catcatgagc ttgaggacaa cttgagctac ataagattcc 13261 catccccacc cccacacaca cacacacatc tagtatgtac ctagtatgca gtcttttcat 13321 atgtttaaaa tgtagaaagg tatatgtgaa gtctcccttt tccccacaac ttagtccctc 13381 caaagtattg catcagtttc cttgtacatg attttatgca tatataagat acactatttc 13441 tctctctctc tctctccttc cctccttccc tccaacctct gggtatgagt tgagggttat 13501 gtatgtttca agttgtaata agaattgcca ctcaaacatt tcatcccatt ttaagttaag 13561 tttgtctgat tttccacttt agaagggcag ccatgatggc tcatgcctat aatcccatcc 13621 tagcacttgt aagcctaata cataagactg gaggattgtc atcagcttga ggctagcctg 13681 gtctacctaa tgagttccag ccagccaaga gtagaatata agaccctgtc tccaaaaaat 13741 gtaccaaagg ggctggaggg gtaactcagt ggttaagaac agtagctgct cttccagagg 13801 acctgggttc aattcccagc acccacatgg cagcttgcaa ctatctgtaa ctgtccagct 13861 ttgggatatc cgacaccatt ataccaatta cataaaatga aaaaattgta aaatgtgcta 13921 aaatgagtgg ggagggggag catggtgtgg ttctgtgggc ttgggatttt gccatgaatc 13981 catgactata tctcagcaat atgtggatta tttctggaaa cctgtggggc atgggtgtat 14041 gggaagaggg gagagagaga gagcccgcat ccagccagag ttcctgtgct ctgggcaggc 14101 ggacgtggga ggactgctgg acgatttcca cttggccctg ggtgggcatc tggctgtgtg 14161 aagccactga ccccactcag cggggggtag ataaggggca tttcccatac caggttctca 14221 gactccagca ttccacgctg gatacagcag aagagctgag aacagaatag gggccccagg 14281 gagacactct cgtcccagag ataaagaagg cagagggaga gagattccta cgcaggcgag 14341 aggccttagt ctgggccttg atgaacagag acggtctatg gttttagagc tttattgtag 14401 aaaggcaggg ggaaagagag aaggtagaaa gagagagacc agccatggcc aagaggagag 14461 aaggggggaa aggagagaga gaagaaaggc tagagagtaa gaaaggttaa agagacagaa 14521 gtaaaagctt agagtaagag agagagagag agcaagaggg agagtaagaa aagtaaaagc 14581 ttaagagagc gaggaggggc cagcccctct aatagtgggc ttgttatctt gttgttgcta 14641 ggtaactgtg aagcgtctag cctgaaggcc agaagcttgg gacattgtct atgtgactga 14701 aagccatgag cctctactgt gggggctgtg gaggtggtaa cttggacagg agccaggggt 14761 tccaggagac atgagggaat gccttctgtc ccatgtaggt aggaattacc accaccggct 14821 ttcaccgtag ttcactggag actgtttgta catagcccat tgccccacag aaaccaggca 14881 ctcactgttt catttgcata agaattagaa ggctttattt tggctgggat tggtgacttg 14941 ccccagaaat tccagctttt gggaagccga atcctggaga accaggagtt caaggtcatc 15001 ctcaggtacc tcatttgttc aagaccagcc tgtgctacca tgagaccttg tccctactcc 15061 cctcaacccc aaaacacgct gttttcattt tataggttgt actaaccatg gaggatggtc 15121 atttgccttt ctcttgattt cagaagcgta gcgcgggctt tgcaccatga tctagtgttt 15181 ccagataggc ttgcctctcc acagccaggc ttatcctgct tcagtcccat gtttggcaag 15241 ggcaacatat gactgaagaa cttgcatagc cttatggtgt atcatgactc ttagccccag 15301 tttataactg tctaacagct ggtggctttt gtgaaaaagg aaagcatcat ttgaactttt 15361 aacttttcag ctgtttgttt tgactttgca taaatactga gactagctag gaaatgagct 15421 tttgtttcat ttgctaaaaa tgattttttt ttcatgaggc aatttcaagc aatttgaatt 15481 tagcaatctc tacttgtctg aaaagaagtt attaattaaa aaaaaacaaa caaacaggaa 15541 cctattatgc tggctagtca catgaaagaa tccaaatgaa cacagtcaag gctgaatcaa 15601 cacatgaaat tattttctaa ttccccaaat tacatttatg ctaattagta tgtagtaaaa 15661 cattttaatt gtttccaatg ttaaacatta cagaaataat tttgatgttt tcctaataac 15721 ccagcctttg atcatgtttg catttatttg attgttgtca gtttattaaa taattaagta 15781 agagcaacaa gataaaatga cttcatgaga gtccttttct gtaacttaga acgtcagtag 15841 tcataggaac tgcggtcgtt cccttacctt tcagaagtag gctaaagagt tgaacgcaaa 15901 ggtggttttt cgaatcttcc tgatccttaa gcccccacta ggtttatatt ctatatttga 15961 tatcggattc gaacccctcg agcttaggat ccggaaccct taatataact aagttattag 16021 gtccctcgaa gaggttcact agtactggcc attgcggccg cttcgaggct gcctcgcgcg 16081 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg 16141 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 16201 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac 16261 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac 16321 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg 16381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 16441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 16501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 16561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 16621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 16681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 16741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 16801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 16861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 16921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 16981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 17041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 17101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 17161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 17221 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 17281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 17341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 17401 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 17461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 17521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 17581 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 17641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 17701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 17761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 17821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 17881 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 17941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 18001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 18061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 18121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 18181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 18241 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt g //