LOCUS Exported 18023 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Target a Cre-dependent GFP and a tdTomato-P2A-tTA2 expression cassette into the mouse TIGRE locus. ACCESSION . VERSION . KEYWORDS Ai139 (TIT2L-GFP-ICL-TPT) targeting vector SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 18023) 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 18023) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..18023 /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 2959..3040 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 2984..3003 /label=SV40pA-R /note="SV40 polyA, reverse primer" protein_bind 3054..3324 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 3357..3395 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3392..3416 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 3529..3562 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 3640..3688 /note="synthetic polyadenylation signal" polyA_signal 3697..4173 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" polyA_signal 4185..4646 /label=PGK poly(A) signal /note="mouse phosphoglycerate kinase 1 polyadenylation signal" protein_bind 4673..4706 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 4720..4729 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4726..5445 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4726..5442 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4771..4792) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5032..5051) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5379..5400 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5470..6058 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(5941..5952) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 6089..6296 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 8826..9129 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 9131..9408 /label=chicken beta-actin promoter intron 9410..10427 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 10351..10373 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 10435..10454 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(10515..10548) /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 10586..10634 /note="synthetic polyadenylation signal" polyA_signal 10643..11119 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(11154..11173) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 11198..11246 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" protein_bind complement(11420..11453) /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)." regulatory 11466..11475 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(11605..11623) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 11873..11892 /label=mCherry-F /note="mCherry, forward primer" primer_bind 12067..12090 /label=DsRed1-C /note="DsRed1, forward primer" CDS 12909..12965 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 12966..13712 /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 13755..14343 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(14226..14237) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 14374..14581 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 14595..14628 /label=attB /bound_moiety="phage phi-C31 integrase" /note="minimal attB site for the phi-C31 integrase (Groth et al., 2000)" promoter 14655..15154 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(14690..14713) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 15087..15106 /label=mPGK-F /note="Mouse PGK promoter, forward primer" primer_bind complement(15802..15818) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(15802..15818) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(15815..15837) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 15826..15842 /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(15850..15880) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 15895..15916 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(16033..16050) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(16204..16792) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(16284..16303) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(16963..17823) /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 17586..17605 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(17824..17928) /gene="bla" /label=AmpR promoter primer_bind 17996..18014 /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 gctgtcccga gctcggtacc gccccatcct cactgactcc gtcctggagt 1741 tggatgagag ataatggcct tacgttgtgc caggggaggg tcgggctgga tttagcaaga 1801 tttaccttct ccaaagagcg gtgctgcagt ggcacagctg cccacggagg tgggggggtc 1861 accgtccctg gaggtgatga agaactgtgg ggatgtggca ctgagggaca tggccagtgg 1921 gcacggtggg tgggttgggg ttggtcttgg ggatcttgga gggcttttcc agccttcatg 1981 atttgacgat tgtatgaaca tctacatggc aattctccag ctgcctgtcc cagtcctact 2041 gacccagctg tatctctcca ggcaagctct tccacccctt ctgcttgcat ccagacacca 2101 tcaaacatgc aggctcagac acagggacca gcagtgtctg tggccttttt gtgctcctct 2161 ccatgctggg ttttaacttg ctctttgtcc ttctatccta tcttcttatc cttaaggctg 2221 ttctgaacgc tgtgacttgg agagtgtccc agagccctca acacctgcat gtcccacgtc 2281 catgctgtcc tgcacttcct tatccccaag atctgcctct ccgtgatgca ctgaattggc 2341 aaacatgtgt caccccagac caacaatgtc acagcaaact cccccttgat aggacaaggg 2401 ggaatggctt tacactgaga caggggaggt ttgggttgga tatgaggagg cagtttttcc 2461 cccagagggt ggtgacgcac tgaacaggtt gcccaaggag gctgtggatg ccccatccct 2521 gcaggcattc aaggccaggc tggatgtggc tctgggcagc ctgggctgct ggttgatgac 2581 cctgcacata gcagggggtt ggatctggat gagcactgtg ctcctttgca acccaggccg 2641 ttctatgatt ctgtcattct aaatctctct ttcagcctaa agctttttcc ccgtatcccc 2701 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc 2761 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc 2821 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat 2881 tacatccctg ggggctttgg gggggggctg tcccggtacc cggggatcca ccggtatcga 2941 ttcggccggc cgacgtcggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat 3001 tgctttattt gtaaccatta taagctgcaa taaacaagtt tcgagtttac cactccctat 3061 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg 3121 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact 3181 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga 3241 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt 3301 accactccct atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgaggta 3361 ggcgtgtacg gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct 3421 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc 3481 gcggccccga attcgagctc ggtacccggg gatcaaattc aattaaatat aacttcgtat 3541 agcatacatt atacgaagtt attcgcgagt gaataaatga aagcttgcag atcccctcag 3601 caccatggct agcggcagcc tcggagtttg aatagataga ataaaatatc tttattttca 3661 ttccatctgt gtgttggttt tttgtgtgag atctacgggt ggcatccctg tgacccctcc 3721 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata 3781 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg 3841 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat 3901 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg 3961 gttcaagcga ttctcctgcc tcaggctccc gagttgttgg gattccaggc atgcatgacc 4021 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg 4081 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta 4141 caggcgtgaa ccactgctcc cttccctgtc cttctgcctc agcgctgtaa gtctgcagaa 4201 attgatgatc tattaaacaa taaagatgtc cactaaaatg gaagtttttc ctgtcatact 4261 ttgttaagaa gggtgagaac agagtaccta cattttgaat ggaaggattg gagctacggg 4321 ggtgggggtg gggtgggatt agataaatgc ctgctcttta ctgaaggctc tttactattg 4381 ctttatgata atgtttcata gttggatatc ataatttaaa caagcaaaac caaattaagg 4441 gccagctcat tcctcccact catgatctat agatctatag atctctcgtg ggatcattgt 4501 ttttctcttg attcccactt tgtggttcta agtactgtgg tttccaaatg tgtcagtttc 4561 atagcctgaa gaacgagatc agcagcctct gttccacata cacttcattc tcagtattgt 4621 tttgccaagt tctaattcca tcagaaagct tttaattaac tgaatagcta acataacttc 4681 gtatagcata cattatacga agttatcacg cgtgctagcg ccaccatggt gagcaagggc 4741 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 4801 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 4861 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 4921 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc 4981 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 5041 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 5101 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 5161 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac 5221 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag 5281 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 5341 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 5401 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaacgcg ttacgtaaag 5461 cttgtcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5521 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5581 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5641 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5701 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5761 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5821 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5881 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5941 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 6001 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat 6061 cgacaccgag cgctgctcga ggcctcgact gtgccttcta gttgccagcc atctgttgtt 6121 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 6181 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 6241 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggacacg 6301 tgggatcctc tagtctagag ccccatcctc actgactccg tcctggagtt ggatgagaga 6361 taatggcctt acgttgtgcc aggggagggt cgggctggat ttagcaagat ttaccttctc 6421 caaagagcgg tgctgcagtg gcacagctgc ccacggaggt gggggggtca ccgtccctgg 6481 aggtgatgaa gaactgtggg gatgtggcac tgagggacat ggccagtggg cacggtgggt 6541 gggttggggt tggtcttggg gatcttggag ggcttttcca gccttcatga tttgacgatt 6601 gtatgaacat ctacatggca attctccagc tgcctgtccc agtcctactg acccagctgt 6661 atctctccag gcaagctctt ccaccccttc tgcttgcatc cagacaccat caaacatgca 6721 ggctcagaca cagggaccag cagtgtctgt ggcctttttg tgctcctctc catgctgggt 6781 tttaacttgc tctttgtcct tctatcctat cttcttatcc ttaaggctgt tctgaacgct 6841 gtgacttgga gagtgtccca gagccctcaa cacctgcatg tcccacgtcc atgctgtcct 6901 gcacttcctt atccccaaga tctgcctctc cgtgatgcac tgaattggca aacatgtgtc 6961 accccagacc aacaatgtca cagcaaactc ccccttgata ggacaagggg gaatggcttt 7021 acactgagac aggggaggtt tgggttggat atgaggaggc agtttttccc ccagagggtg 7081 gtgacgcact gaacaggttg cccaaggagg ctgtggatgc cccatccctg caggcattca 7141 aggccaggct ggatgtggct ctgggcagcc tgggctgctg gttgatgacc ctgcacatag 7201 cagggggttg gatctggatg agcactgtgc tcctttgcaa cccaggccgt tctatgattc 7261 tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc caggtgtctg 7321 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt 7381 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc 7441 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg 7501 gggctttggg ggggggctgt ccctctagag ccccatcctc actgactccg tcctggagtt 7561 ggatgagaga taatggcctt acgttgtgcc aggggagggt cgggctggat ttagcaagat 7621 ttaccttctc caaagagcgg tgctgcagtg gcacagctgc ccacggaggt gggggggtca 7681 ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac tgagggacat ggccagtggg 7741 cacggtgggt gggttggggt tggtcttggg gatcttggag ggcttttcca gccttcatga 7801 tttgacgatt gtatgaacat ctacatggca attctccagc tgcctgtccc agtcctactg 7861 acccagctgt atctctccag gcaagctctt ccaccccttc tgcttgcatc cagacaccat 7921 caaacatgca ggctcagaca cagggaccag cagtgtctgt ggcctttttg tgctcctctc 7981 catgctgggt tttaacttgc tctttgtcct tctatcctat cttcttatcc ttaaggctgt 8041 tctgaacgct gtgacttgga gagtgtccca gagccctcaa cacctgcatg tcccacgtcc 8101 atgctgtcct gcacttcctt atccccaaga tctgcctctc cgtgatgcac tgaattggca 8161 aacatgtgtc accccagacc aacaatgtca cagcaaactc ccccttgata ggacaagggg 8221 gaatggcttt acactgagac aggggaggtt tgggttggat atgaggaggc agtttttccc 8281 ccagagggtg gtgacgcact gaacaggttg cccaaggagg ctgtggatgc cccatccctg 8341 caggcattca aggccaggct ggatgtggct ctgggcagcc tgggctgctg gttgatgacc 8401 ctgcacatag cagggggttg gatctggatg agcactgtgc tcctttgcaa cccaggccgt 8461 tctatgattc tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc 8521 caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca 8581 ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc 8641 ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt 8701 acatccctgg gggctttggg ggggggctgt ccctctagaa gtcgacggcg cgccctgcag 8761 gaatctagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 8821 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 8881 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 8941 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 9001 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 9061 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 9121 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc 9181 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg 9241 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg 9301 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg 9361 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg gagtcgctgc 9421 gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct 9481 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta 9541 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg 9601 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt 9661 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg 9721 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc 9781 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg 9841 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc 9901 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc 9961 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc 10021 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag 10081 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc 10141 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg 10201 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg 10261 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg 10321 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct 10381 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg 10441 tgctggttat tgtgctgtct catcattttg gcaaagaatt gatttgatac cgcgggcccc 10501 gcgtggacgg taccataact tcgtataaag tatcctatac gaagttatcc atggctagcg 10561 gcagcctcgg agtttgaata gatagaataa aatatcttta ttttcattcc atctgtgtgt 10621 tggttttttg tgtgagatct acgggtggca tccctgtgac ccctccccag tgcctctcct 10681 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat 10741 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag 10801 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg 10861 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct 10921 cctgcctcag gctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt 10981 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta 11041 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac 11101 tgctcccttc cctgtccttc tgcctcagct aattgagtag gggggaggct aactgaaaca 11161 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 11221 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc 11281 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc 11341 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg 11401 cctcgcgagg gtttctagaa taacttcgta taaagtatcc tatacgaagt tatacgcgta 11461 ctagcgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg 11521 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc 11581 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct 11641 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc 11701 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg 11761 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg 11821 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa 11881 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg 11941 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg 12001 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg 12061 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc 12121 gctccgaggg ccgccaccac ctgttcctgg ggcatggtac cggcagcacc ggcagcggca 12181 gctccggcac cgcctcctcc gaggacaaca acatggccgt gattaaggag tttatgaggt 12241 ttaaagtcag gatggagggc agcatgaacg ggcacgagtt tgagattgaa ggagaggggg 12301 aagggaggcc ctatgagggc acccagacag caaagctgaa agtgacaaag ggcggccctc 12361 tgccattcgc atgggacatc ctgtctcctc agtttatgta tggaagtaaa gcctacgtga 12421 aacaccctgc tgacattcca gattacaaga aactgagctt cccagagggg tttaagtggg 12481 aaagagtgat gaacttcgaa gacgggggcc tggtgactgt cacccaggac agctctctgc 12541 aggacggcac tctgatctat aaggtgaaaa tgcgcggcac caattttccc cctgatggcc 12601 cagtcatgca gaagaagaca atgggctggg aggcttcaac agaaaggctg taccccagag 12661 acggcgtgct gaagggagag atccaccagg cactgaagct gaaagatgga gggcattatc 12721 tggtcgaatt caaaaccatc tacatggcca agaaacccgt gcagctgcct ggctactatt 12781 acgtcgacac aaagctggat attactagcc ataacgaaga ctacaccatt gtggagcagt 12841 atgagcggag cgaggggagg caccatctgt ttctgtacgg catggacgag ctgtacaagg 12901 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg 12961 gacctatgtc tagactggac aagagcaaag tcataaactc tgctctggaa ttactcaatg 13021 aagtcggtat cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc 13081 ctaccctgta ctggcacgtg aagaacaagc gggccctgct cgatgccctg gcaatcgaga 13141 tgctggacag gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc 13201 tgcggaacaa cgccaagtca ttccgctgtg ctctcctctc acatcgcgac ggggctaaag 13261 tgcatctcgg cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt 13321 tcctgtgtca gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc 13381 actttacact gggctgcgta ttggaggatc aggagcatca agtagcaaaa gaggaaagag 13441 agacacctac caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc 13501 atcagggagc cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga 13561 aacagctaaa gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca 13621 tgctcccagc cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg 13681 acgattttga ccttgacatg ctccccgggt aactaagtaa ggatcaaatg gcgcgttacg 13741 taaagcttgt cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 13801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 13861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 13921 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 13981 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 14041 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 14101 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct 14161 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg 14221 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 14281 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 14341 cgcatcgaca ccgagcgctg ctcgaggcct cgactgtgcc ttctagttgc cagccatctg 14401 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 14461 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 14521 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 14581 acacgtggga tccggtgcca gggcgtgccc ttgggctccc cgggcgcgtt tgggcggccg 14641 catcgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 14701 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 14761 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 14821 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 14881 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 14941 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 15001 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 15061 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 15121 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc aatcgccgct agccgccacc 15181 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac 15241 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat 15301 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat 15361 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt 15421 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg 15481 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat 15541 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga 15601 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat 15661 cactggcaaa ctgtgatgga cgacaccgtc agtgtaagtg ctagccccta ggcctgaatc 15721 gaagttccta ttcttcaaaa ggtataggaa cttcgagctc gtcgactgca gaggcctgca 15781 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 15841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 15901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 15961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 16021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 16081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 16141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 16201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 16261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 16321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 16381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 16441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 16501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 16561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 16621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 16681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 16741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 16801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 16861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 16921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 16981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 17041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 17101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 17161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 17221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 17281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 17341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 17401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 17461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 17521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 17581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 17641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 17701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 17761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 17821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 17881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 17941 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 18001 gcgtatcacg aggccctttc gtc //