LOCUS Exported 18320 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Target a Cre-dependent ChrimsonR-tdTomato cassette and a tTA2 cassette into the mouse TIGRE locus. ACCESSION . VERSION . KEYWORDS Ai167 (TIT2L-ChrimsonR-tdT-ICL-tTA2) targeting vector SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 18320) AUTHORS Daigle TL, Madisen L, Hage TA, Valley MT, Knoblich U, Larsen RS, Takeno MM, Huang L, Gu H, Larsen R, Mills M, Bosma-Moody A, Siverts LA, Walker M, Graybuck LT, Yao Z, Fong O, Nguyen TN, Garren E, Lenz GH, Chavarha M, Pendergraft J, Harrington J, Hirokawa KE, Harris JA, Nicovich PR, McGraw MJ, Ollerenshaw DR, Smith KA, Baker CA, Ting JT, Sunkin SM, Lecoq J, Lin MZ, Boyden ES, Murphy GJ, da Costa NM, Waters J, Li L, Tasic B, Zeng H TITLE A Suite of Transgenic Driver and Reporter Mouse Lines with Enhanced Brain-Cell-Type Targeting and Functionality. JOURNAL Cell. 2018 Jul 12;174(2):465-480.e22. doi: 10.1016/j.cell.2018.06.035. PUBMED 30007418 REFERENCE 2 (bases 1 to 18320) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..18320 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 2975..3056 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 3000..3019 /label=SV40pA-R /note="SV40 polyA, reverse primer" protein_bind 3070..3340 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 3373..3411 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3408..3432 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 3545..3578 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 3655..3703 /note="synthetic polyadenylation signal" polyA_signal 3712..4188 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" polyA_signal 4200..4661 /label=PGK poly(A) signal /note="mouse phosphoglycerate kinase 1 polyadenylation signal" protein_bind 4688..4721 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 4732..4741 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4738..5787 /codon_start=1 /product="red light-drivable channelrhodopsin from Chlamydomonas noctigama (Klapoetke et al., 2014)" /label=ChrimsonR /note="K176R mutant with accelerated closing kinetics" /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE FVEEEDEDTV" misc_feature 7258..7846 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(7729..7740) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 7877..8084 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 10608..10911 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 10913..11190 /label=chicken beta-actin promoter intron 11191..12207 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 12131..12153 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 12215..12234 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(12295..12328) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." polyA_signal 12366..12414 /note="synthetic polyadenylation signal" polyA_signal 12423..12899 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(12934..12953) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 12978..13026 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" protein_bind complement(13205..13238) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." CDS 13263..14009 /codon_start=1 /product="improved tetracycline-controlled transactivator" /label=tTA-Advanced /note="In the Tet-Off(R) system, tTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the absence of tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" misc_feature 14052..14640 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(14523..14534) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 14671..14878 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 14892..14925 /label=attB /bound_moiety="phage phi-C31 integrase" /note="minimal attB site for the phi-C31 integrase (Groth et al., 2000)" promoter 14952..15451 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(14987..15010) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 15384..15403 /label=mPGK-F /note="Mouse PGK promoter, forward primer" primer_bind complement(16099..16115) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(16099..16115) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(16112..16134) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 16123..16139 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(16147..16177) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 16192..16213 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(16330..16347) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(16501..17089) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(16581..16600) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(17260..18120) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 17883..17902 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(18121..18225) /gene="bla" /label=AmpR promoter primer_bind 18293..18311 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accgaagttc 421 ctattcttca aatagtatag gaacttcttt cggccggcca atcgattgga tccgtttaat 481 tcgagctcgc tagagcccca tcctcactga ctccgtcctg gagttggatg agagataatg 541 gccttacgtt gtgccagggg agggtcgggc tggatttagc aagatttacc ttctccaaag 601 agcggtgctg cagtggcaca gctgcccacg gaggtggggg ggtcaccgtc cctggaggtg 661 atgaagaact gtggggatgt ggcactgagg gacatggcca gtgggcacgg tgggtgggtt 721 ggggttggtc ttggggatct tggagggctt ttccagcctt catgatttga cgattgtatg 781 aacatctaca tggcaattct ccagctgcct gtcccagtcc tactgaccca gctgtatctc 841 tccaggcaag ctcttccacc ccttctgctt gcatccagac accatcaaac atgcaggctc 901 agacacaggg accagcagtg tctgtggcct ttttgtgctc ctctccatgc tgggttttaa 961 cttgctcttt gtccttctat cctatcttct tatccttaag gctgttctga acgctgtgac 1021 ttggagagtg tcccagagcc ctcaacacct gcatgtccca cgtccatgct gtcctgcact 1081 tccttatccc caagatctgc ctctccgtga tgcactgaat tggcaaacat gtgtcacccc 1141 agaccaacaa tgtcacagca aactccccct tgataggaca agggggaatg gctttacact 1201 gagacagggg aggtttgggt tggatatgag gaggcagttt ttcccccaga gggtggtgac 1261 gcactgaaca ggttgcccaa ggaggctgtg gatgccccat ccctgcaggc attcaaggcc 1321 aggctggatg tggctctggg cagcctgggc tgctggttga tgaccctgca catagcaggg 1381 ggttggatct ggatgagcac tgtgctcctt tgcaacccag gccgttctat gattctgtca 1441 ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc 1501 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg 1561 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc 1621 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct 1681 ttgggggggg gctgtccctc tagcgagctc ggtacccaga gccccatcct cactgactcc 1741 gtcctggagt tggatgagag ataatggcct tacgttgtgc caggggaggg tcgggctgga 1801 tttagcaaga tttaccttct ccaaagagcg gtgctgcagt ggcacagctg cccacggagg 1861 tgggggggtc accgtccctg gaggtgatga agaactgtgg ggatgtggca ctgagggaca 1921 tggccagtgg gcacggtggg tgggttgggg ttggtcttgg ggatcttgga gggcttttcc 1981 agccttcatg atttgacgat tgtatgaaca tctacatggc aattctccag ctgcctgtcc 2041 cagtcctact gacccagctg tatctctcca ggcaagctct tccacccctt ctgcttgcat 2101 ccagacacca tcaaacatgc aggctcagac acagggacca gcagtgtctg tggccttttt 2161 gtgctcctct ccatgctggg ttttaacttg ctctttgtcc ttctatccta tcttcttatc 2221 cttaaggctg ttctgaacgc tgtgacttgg agagtgtccc agagccctca acacctgcat 2281 gtcccacgtc catgctgtcc tgcacttcct tatccccaag atctgcctct ccgtgatgca 2341 ctgaattggc aaacatgtgt caccccagac caacaatgtc acagcaaact cccccttgat 2401 aggacaaggg ggaatggctt tacactgaga caggggaggt ttgggttgga tatgaggagg 2461 cagtttttcc cccagagggt ggtgacgcac tgaacaggtt gcccaaggag gctgtggatg 2521 ccccatccct gcaggcattc aaggccaggc tggatgtggc tctgggcagc ctgggctgct 2581 ggttgatgac cctgcacata gcagggggtt ggatctggat gagcactgtg ctcctttgca 2641 acccaggccg ttctatgatt ctgtcattct aaatctctct ttcagcctaa agctttttcc 2701 ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg 2761 atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg 2821 gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga 2881 gggacgtaat tacatccctg ggggctttgg gggggggctg tccctctagg ggtacccggg 2941 gatccaccgg tatcgattcg gccggccgac gtcggcagtg aaaaaaatgc tttatttgtg 3001 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagtttcga 3061 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca 3121 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa 3181 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc 3241 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa 3301 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg 3361 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 3421 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 3481 gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc aaattcaatt 3541 aaatataact tcgtatagca tacattatac gaagttattc gcgatgaata aatgaaagct 3601 tgcagatccc ctcagcacca tggctagcgg cagcctcgga gtttgaatag atagaataaa 3661 atatctttat tttcattcca tctgtgtgtt ggttttttgt gtgagatcta cgggtggcat 3721 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc 3781 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat 3841 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg 3901 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca 3961 atctccgcct cctgggttca agcgattctc ctgcctcagg ctcccgagtt gttgggattc 4021 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc 4081 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc 4141 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gcctcagcgc 4201 tgtaagtctg cagaaattga tgatctatta aacaataaag atgtccacta aaatggaagt 4261 ttttcctgtc atactttgtt aagaagggtg agaacagagt acctacattt tgaatggaag 4321 gattggagct acgggggtgg gggtggggtg ggattagata aatgcctgct ctttactgaa 4381 ggctctttac tattgcttta tgataatgtt tcatagttgg atatcataat ttaaacaagc 4441 aaaaccaaat taagggccag ctcattcctc ccactcatga tctatagatc tatagatctc 4501 tcgtgggatc attgtttttc tcttgattcc cactttgtgg ttctaagtac tgtggtttcc 4561 aaatgtgtca gtttcatagc ctgaagaacg agatcagcag cctctgttcc acatacactt 4621 cattctcagt attgttttgc caagttctaa ttccatcaga aagcttttaa ttaactgaat 4681 agctaacata acttcgtata gcatacatta tacgaagtta tcacgcgtgc cgccaccatg 4741 gctgagctga tcagcagcgc caccagatct ctgtttgccg ccggaggcat caacccttgg 4801 cctaacccct accaccacga ggacatgggc tgtggaggaa tgacacctac aggcgagtgc 4861 ttcagcaccg agtggtggtg tgacccttct tacggactga gcgacgccgg atacggatat 4921 tgcttcgtgg aggccacagg cggctacctg gtcgtgggag tggagaagaa gcaggcttgg 4981 ctgcacagca gaggcacacc aggagaaaag atcggcgccc aggtctgcca gtggattgct 5041 ttcagcatcg ccatcgccct gctgacattc tacggcttca gcgcctggaa ggccacttgc 5101 ggttgggagg aggtctacgt ctgttgcgtc gaggtgctgt tcgtgaccct ggagatcttc 5161 aaggagttca gcagccccgc cacagtgtac ctgtctaccg gcaaccacgc ctattgcctg 5221 cgctacttcg agtggctgct gtcttgcccc gtgatcctga tcagactgag caacctgagc 5281 ggcctgaaga acgactacag caagcggacc atgggcctga tcgtgtcttg cgtgggaatg 5341 atcgtgttcg gcatggccgc aggactggct accgattggc tcaagtggct gctgtatatc 5401 gtgtcttgca tctacggcgg ctacatgtac ttccaggccg ccaagtgcta cgtggaagcc 5461 aaccacagcg tgcctaaagg ccattgccgc atggtcgtga agctgatggc ctacgcttac 5521 ttcgcctctt ggggcagcta cccaatcctc tgggcagtgg gaccagaagg actgctgaag 5581 ctgagccctt acgccaacag catcggccac agcatctgcg acatcatcgc caaggagttt 5641 tggaccttcc tggcccacca cctgaggatc aagatccacg agcacatcct gatccacggc 5701 gacatccgga agaccaccaa gatggagatc ggaggcgagg aggtggaagt ggaagagttc 5761 gtggaggagg aggacgagga cacagtggcg gcaccggtag tagcagtgag taagggcgag 5821 gaagtgatca aagagttcat gcggtttaag gtgagaatgg aaggaagcat gaacggccac 5881 gagttcgaaa ttgagggaga aggagaggga cggccctacg agggcaccca gacagccaag 5941 ctgaaagtga caaagggcgg gcctctgcca ttcgcttggg acatcctgag cccacagttt 6001 atgtacggct ccaaggccta tgtgaaacat ccagctgaca ttcccgatta taagaaactg 6061 agcttccccg aggggtttaa gtgggaaaga gtgatgaact tcgaggacgg aggcctggtg 6121 actgtgaccc aggacagctc cctgcaggat gggaccctga tctacaaggt gaaaatgaga 6181 gggacaaatt ttccccctga tggacctgtg atgcagaaga aaactatggg atgggaggcc 6241 tccaccgaaa ggctgtatcc acgcgacggg gtgctgaaag gagaaatcca ccaggctctg 6301 aagctgaaag atgggggaca ttacctggtg gagttcaaga caatctacat ggccaagaaa 6361 cctgtgcagc tgccaggcta ctattacgtg gacacaaaac tggatatcac ttcacacaac 6421 gaggactaca ctattgtgga gcagtatgaa cggagcgagg ggagacacca tctgttcctg 6481 ggccatggga ctggaagtac cggctcaggg tctagtggaa ccgcctcaag cgaggataac 6541 aatatggctg tgatcaaaga gttcatgagg tttaaggtgc gcatggaggg cagcatgaat 6601 gggcacgaat ttgagattga aggagagggc gaagggaggc cttacgaggg cacacagact 6661 gccaagctga aagtgaccaa gggaggacca ctgcctttcg cttgggatat cctgtctcct 6721 cagtttatgt acggaagtaa ggcctatgtc aagcatcccg ctgacattcc tgattacaag 6781 aaactgtctt tcccagaggg ctttaagtgg gagagagtga tgaattttga agatggaggc 6841 ctggtgaccg tgacacagga ctcctctctg caggatggca ctctgatcta caaagtcaaa 6901 atgcgcggca ccaattttcc acccgatggg cccgtgatgc agaagaaaac aatggggtgg 6961 gaggccagca ctgaacggct gtatcctaga gacggagtgc tgaagggcga aatccaccag 7021 gccctgaagc tgaaagacgg cggccactac ctggtggagt tcaaaaccat ctacatggcc 7081 aagaaaccag tgcagctgcc cggctattac tatgtggaca ccaagctgga tatcacatcc 7141 cacaatgaag actacaccat tgtggaacag tatgagaggt ctgaaggacg ccaccatctg 7201 tttctgtacg gcatggatga gctgtataag taaacgcgtt acgtaaagct tgtcgataat 7261 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 7321 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 7381 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 7441 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 7501 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 7561 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 7621 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc 7681 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 7741 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 7801 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg acaccgagcg 7861 ctgctcgagg cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 7921 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 7981 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 8041 agcaaggggg aggattggga agacaatagc aggcatgctg gggacacgtg ggatcctcta 8101 gagccccatc ctcactgact ccgtcctgga gttggatgag agataatggc cttacgttgt 8161 gccaggggag ggtcgggctg gatttagcaa gatttacctt ctccaaagag cggtgctgca 8221 gtggcacagc tgcccacgga ggtggggggg tcaccgtccc tggaggtgat gaagaactgt 8281 ggggatgtgg cactgaggga catggccagt gggcacggtg ggtgggttgg ggttggtctt 8341 ggggatcttg gagggctttt ccagccttca tgatttgacg attgtatgaa catctacatg 8401 gcaattctcc agctgcctgt cccagtccta ctgacccagc tgtatctctc caggcaagct 8461 cttccacccc ttctgcttgc atccagacac catcaaacat gcaggctcag acacagggac 8521 cagcagtgtc tgtggccttt ttgtgctcct ctccatgctg ggttttaact tgctctttgt 8581 ccttctatcc tatcttctta tccttaaggc tgttctgaac gctgtgactt ggagagtgtc 8641 ccagagccct caacacctgc atgtcccacg tccatgctgt cctgcacttc cttatcccca 8701 agatctgcct ctccgtgatg cactgaattg gcaaacatgt gtcaccccag accaacaatg 8761 tcacagcaaa ctcccccttg ataggacaag ggggaatggc tttacactga gacaggggag 8821 gtttgggttg gatatgagga ggcagttttt cccccagagg gtggtgacgc actgaacagg 8881 ttgcccaagg aggctgtgga tgccccatcc ctgcaggcat tcaaggccag gctggatgtg 8941 gctctgggca gcctgggctg ctggttgatg accctgcaca tagcaggggg ttggatctgg 9001 atgagcactg tgctcctttg caacccaggc cgttctatga ttctgtcatt ctaaatctct 9061 ctttcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc 9121 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc 9181 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc 9241 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc 9301 tgtccctcta gagccccatc ctcactgact ccgtcctgga gttggatgag agataatggc 9361 cttacgttgt gccaggggag ggtcgggctg gatttagcaa gatttacctt ctccaaagag 9421 cggtgctgca gtggcacagc tgcccacgga ggtggggggg tcaccgtccc tggaggtgat 9481 gaagaactgt ggggatgtgg cactgaggga catggccagt gggcacggtg ggtgggttgg 9541 ggttggtctt ggggatcttg gagggctttt ccagccttca tgatttgacg attgtatgaa 9601 catctacatg gcaattctcc agctgcctgt cccagtccta ctgacccagc tgtatctctc 9661 caggcaagct cttccacccc ttctgcttgc atccagacac catcaaacat gcaggctcag 9721 acacagggac cagcagtgtc tgtggccttt ttgtgctcct ctccatgctg ggttttaact 9781 tgctctttgt ccttctatcc tatcttctta tccttaaggc tgttctgaac gctgtgactt 9841 ggagagtgtc ccagagccct caacacctgc atgtcccacg tccatgctgt cctgcacttc 9901 cttatcccca agatctgcct ctccgtgatg cactgaattg gcaaacatgt gtcaccccag 9961 accaacaatg tcacagcaaa ctcccccttg ataggacaag ggggaatggc tttacactga 10021 gacaggggag gtttgggttg gatatgagga ggcagttttt cccccagagg gtggtgacgc 10081 actgaacagg ttgcccaagg aggctgtgga tgccccatcc ctgcaggcat tcaaggccag 10141 gctggatgtg gctctgggca gcctgggctg ctggttgatg accctgcaca tagcaggggg 10201 ttggatctgg atgagcactg tgctcctttg caacccaggc cgttctatga ttctgtcatt 10261 ctaaatctct ctttcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc 10321 aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg 10381 ctgtccccgc acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc 10441 cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt 10501 gggggggggc tgtccctcta gagtcgacgg cgcgccctgc aggaatctag ttattaatag 10561 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 10621 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 10681 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 10741 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 10801 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 10861 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg 10921 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat 10981 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg 11041 ccrggsgggg sggggsgggg cgrggggsgg ggcggggcga ggcggagagg tgcggcggca 11101 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg 11161 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg 11221 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca 11281 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga 11341 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg 11401 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc 11461 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc 11521 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc 11581 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc 11641 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg 11701 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg 11761 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg 11821 gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat 11881 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg 11941 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc 12001 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc 12061 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg 12121 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat 12181 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct 12241 catcattttg gcaaagaatt gatttgatac cgcgggcccc gcgtggacgg taccataact 12301 tcgtataaag tatcctatac gaagttatcc atggctagcg gcagcctcgg agtttgaata 12361 gatagaataa aatatcttta ttttcattcc atctgtgtgt tggttttttg tgtgagatct 12421 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc 12481 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt 12541 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga 12601 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt 12661 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag gctcccgagt 12721 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac 12781 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac 12841 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc 12901 tgcctcagct aattgagtag gggggaggct aactgaaaca cggaaggaga caataccgga 12961 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt 13021 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 13081 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 13141 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccgtggaagt tcctattctc 13201 tagaataact tcgtataaag tatcctatac gaagttatac gcgcgcgtcc atttaattca 13261 ccatgtctag actggacaag agcaaagtca taaactctgc tctggaatta ctcaatgaag 13321 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta 13381 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctggca atcgagatgc 13441 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc 13501 ggaacaacgc caagtcattc cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc 13561 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc 13621 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact 13681 ttacactggg ctgcgtattg gaggatcagg agcatcaagt agcaaaagag gaaagagaga 13741 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccatc 13801 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac 13861 agctaaagtg cgaaagcggc gggccggccg acgcccttga cgattttgac ttagacatgc 13921 tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg 13981 attttgacct tgacatgctc cccgggtaac taagtaagga tcaaatggcg cgttacgtaa 14041 agcttgtcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 14101 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 14161 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 14221 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 14281 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 14341 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 14401 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 14461 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 14521 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 14581 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 14641 atcgacaccg agcgctgctc gaggcctcga ctgtgccttc tagttgccag ccatctgttg 14701 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct 14761 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg 14821 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaca 14881 cgtgggatcc ggtgccaggg cgtgcccttg ggctccccgg gcgcgtttgg gcggccgcat 14941 cgaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 15001 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 15061 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 15121 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 15181 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 15241 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 15301 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 15361 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 15421 gttctcctct tcctcatctc cgggcctttc gacctgcaat cgccgctagc cgccaccatg 15481 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc 15541 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta 15601 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt 15661 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg 15721 gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa 15781 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg 15841 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc 15901 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac 15961 tggcaaactg tgatggacga caccgtcagt gtaagtgcta gcccctaggc ctgaatcgaa 16021 gttcctattc ttcaaaaggt ataggaactt cgagctcgtc gactgcagag gcctgcatgc 16081 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 16141 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 16201 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 16261 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 16321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 16381 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 16441 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 16501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 16561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 16621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 16681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 16741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 16801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 16861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 16921 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 16981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 17041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 17101 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 17161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 17221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 17281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 17341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 17401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 17461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 17521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 17581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 17641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 17701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 17761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 17821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 17881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 17941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 18001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 18061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 18121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 18181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 18241 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 18301 tatcacgagg ccctttcgtc //