LOCUS Exported 13284 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Donor construct for introduction of hNIL factors to CLYBL safe harbor site and iPSC differentiation to motor neuron. ACCESSION . VERSION . KEYWORDS pUCM-CLYBL-hNIL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13284) AUTHORS Fernandopulle MS, Prestil R, Grunseich C, Wang C, Gan L, Ward ME TITLE Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons. JOURNAL Curr Protoc Cell Biol. 2018 Jun;79(1):e51. doi: 10.1002/cpcb.51. Epub 2018 May 18. PUBMED 29924488 REFERENCE 2 (bases 1 to 13284) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13284 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 266..285 /label=pBR322ori-F /note="pBR322 origin, forward primer" protein_bind 1391..1409 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1427..1445 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1463..1481 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1499..1517 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1535..1553 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1571..1589 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1607..1625 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 1670..1694 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2614..2670 /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" regulatory 3461..3470 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3742..3795 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" polyA_signal 5027..5148 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5064..5083) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5118..5137 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5197..5213) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5197..5213) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5210..5232) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5221..5237 /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(5245..5275) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5290..5311 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5428..5445) /label=L4440 /note="L4440 vector, forward primer" primer_bind 5737..5756 /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" polyA_signal 5755..5810 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind 5856..5875 /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" CDS complement(5965..6711) /codon_start=1 /product="modified rtTA protein that binds tightly to promoters containing the tet operator in the presence of doxycycline" /label=Tet-On(R) 3G /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" primer_bind complement(6743..6762) /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" intron 6770..7787 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind complement(6824..6846) /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" promoter complement(7789..8066) /label=chicken beta-actin promoter enhancer 8068..8447 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" polyA_signal 8453..8677 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 8666..8683 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" protein_bind complement(8699..8732) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(9008..9027) /label=mCherry-F /note="mCherry, forward primer" primer_bind 9277..9295 /label=mCherry-R /note="mCherry, reverse primer" regulatory 9437..9446 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" LTR 9484..9752 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" promoter complement(9765..9976) /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" polyA_signal 10012..10236 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" CDS complement(10241..11032) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(10352..10371) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 10962..10981 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(11036..11089) /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" misc_feature 11113..11138 /label=SA /note="splice acceptor site" protein_bind complement(11139..11172) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 12030..12890 /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 complement(12248..12267) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 13061..365 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 61 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc gaattcgccc ttagagcaga 421 gccaggaacc cctgtaggga aggggcagga gagccagggg catgagatgg tggacgagga 481 agggggacag ggaagcctga gcgcctctcc tgggcttgcc aaggactcaa acccagaagc 541 ccagagcagg gccttaggga agcgggacgg ctctgttgga gagtccagta ttgaattagt 601 gatagataca cagaaaaaaa gcaaattgaa taggtgaatg gataaacaaa ctgtggaaca 661 tccagacaat gaaatattca ccaataaaaa gaaatgagct atcaagccac attggctatg 721 cccttcacat ttaaggagga accttaaatg catattgcca aattttaaaa gctagtctga 781 aaatgctaca gactgtatga ttccaattat atgacatttt agaaaaggca aaactataga 841 gatcagtggt tgccagcggc tcatggaggc aggatgaatt ggtggaacac agggatgctt 901 tcagggtaat gaaaatatct tgtgtgatcc tgtaataatg gatacgtggc ataatgcatt 961 tgttaaaacc tgtaaactag ccaggtatag tggctttttt gcacctatag tctcagctac 1021 ttggtaggct gagacaggaa gatcacttga gcccaggact tcaagaccag cctgggtaat 1081 atagtaagac cccatctcta ttttttaaag aaattaagaa agaaaagaaa cccatcaact 1141 gtataacaca aagaatcgta atgtaaacta cagtcctgag taatgaaaca tttttaaaga 1201 aataaataaa ttcaaggcaa ttattaactt caggaaaaac agtaaagtaa acataagtat 1261 acaaatcaaa catggctcag ttgtgaaaat atttacaagg tcactttaag taacattgaa 1321 tgttgatata atcccaaaat atatttatgt tggaaggatg aggagatatc tccaccccac 1381 agtggtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat 1441 agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc 1501 cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc 1561 tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat 1621 agagaacgta taagctttag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag 1681 tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatacc aactttccgt 1741 accacttcct accctcgtaa acttaaggtt aattaagccg ccaccatgtt cgtcaaatcc 1801 gagaccttgg agttgaagga ggaagaggac gtgttagtgc tgctcggatc ggcctccccc 1861 gccttggcgg ccctgacccc gctgtcatcc agcgccgacg aagaagagga ggaggagccg 1921 ggcgcgtcag gcggggcgcg tcggcagcgc ggggctgagg ccgggcaggg ggcgcggggc 1981 ggcgtggctg cgggtgcgga gggctgccgg cccgcacggc tgctgggtct ggtacacgat 2041 tgcaaacggc gcccttcccg ggcgcgggcc gtctcccgag gcgccaagac ggccgagacg 2101 gtgcagcgca tcaagaagac ccgtagactg aaggccaaca accgcgagcg aaaccgcatg 2161 cacaacctca acgcggcact ggacgcgctg cgcgaggtgc tccccacgtt ccccgaggac 2221 gccaagctca ccaagatcga gaccctgcgc ttcgcccaca actacatctg ggcactcacc 2281 gagaccctgc gcctggcgga tcactgcggg ggcggcggcg ggggcctgcc gggggcgctc 2341 ttctccgagg cagtgttgct gagcccggga ggagccagcg ccgccctgag cagcagcgga 2401 gacagcccct cgcccgcctc cacgtggagt tgcaccaaca gccccgcgcc gtcctcctcc 2461 gtgtcctcca attccacctc cccctacagc tgcactttat cgcccgccag cccggccggg 2521 tcagacatgg actattggca gcccccacct cccgacaagc accgctatgc acctcacctc 2581 cccatagcca gggattgtat cttgggatcc ggagctacta acttcagcct gctgaagcag 2641 gctggagacg tggaggagaa ccctggacct atgggagaca tgggagatcc accaaaaaaa 2701 aaacgtctga tttccctatg tgttggttgc ggcaatcaga ttcacgatca gtatattctg 2761 agggtttctc cggatttgga atggcatgcg gcatgtttga aatgtgcgga gtgtaatcag 2821 tatttggacg agagctgtac atgctttgtt agggatggga aaacctactg taaaagagat 2881 tatatcaggt tgtacgggat caaatgcgcc aagtgcagca tcggcttcag caagaacgac 2941 ttcgtgatgc gtgcccgctc caaggtgtat cacatcgagt gtttccgctg tgtggcctgc 3001 agccgccagc tcatccctgg ggacgaattt gcgcttcggg aggacggtct cttctgccga 3061 gcagaccacg atgtggtgga gagggccagt ctaggcgctg gcgacccgct cagtcccctg 3121 catccagcgc ggccactgca aatggcagcg gagcccatct ccgccaggca gccagccctg 3181 cggccccacg tccacaagca gccggagaag accacccgcg tgcggactgt gctgaacgag 3241 aagcagctgc acaccttgcg gacctgctac gccgcaaacc cgcggccaga tgcgctcatg 3301 aaggagcaac tggtagagat gacgggcctc agtccccgtg tgatccgggt ctggtttcaa 3361 aacaagcggt gcaaggacaa gaagcgaagc atcatgatga agcaactcca gcagcagcag 3421 cccaatgaca aaactaatat ccaggggatg acaggaactc ccatggtggc tgccagtcca 3481 gagagacacg acggtggctt acaggctaac ccagtggaag tacaaagtta ccagccacct 3541 tggaaagtac tgagcgactt cgccttgcag agtgacatag atcagcctgc ttttcagcaa 3601 ctggtcaatt tttcagaagg aggaccgggc tctaattcca ctggcagtga agtagcatca 3661 atgtcctctc aacttccaga tacacctaac agcatggtag ccagtcctat tgaggcattg 3721 atgcacagca gaggctctgg cgagggcaga ggcagcctgc tgacatgtgg cgacgtggaa 3781 gagaaccctg gccccggtac catggaagcc agaggcgaac tgggccctgc cagagaatct 3841 gccggcggag atctgctgct ggccctgctg gctagaaggg ccgacctgag aagagaaatc 3901 cccctgtgcg ccggctgcga ccagcacatc ctggaccggt ttatcctgaa ggccctggac 3961 agacactggc acagcaagtg cctgaagtgc agcgactgcc acacccccct ggccgagaga 4021 tgctttagca gaggcgagag cgtgtactgc aaggacgact tcttcaagag attcggcacc 4081 aagtgcgccg cctgccagct gggaattcct cctacacagg tcgtgcggag agcccaggac 4141 ttcgtgtacc atctgcactg cttcgcctgc gtcgtgtgca agagacagct ggccaccggc 4201 gacgagttct acctgatgga agatagccgg ctcgtgtgta aagccgacta cgagacagcc 4261 aagcagagag aggccgaggc caccgccaaa agacccagaa ccaccatcac cgccaagcag 4321 ctggaaaccc tgaagtccgc ctacaacacc agccccaagc ccgccagaca tgtgcgggaa 4381 cagctgagca gcgagacagg cctggacatg agagtggtgc aagtgtggtt ccagaacaga 4441 cgggccaaag agaagcggct gaagaaggac gccggcaggc agagatgggg ccagtacttc 4501 cggaacatga agcggtccag aggcggcagc aagagcgaca aggacagcgt gcaggaaggc 4561 caggacagcg acgccgaggt gtccttccca gatgagccta gcctggccga aatgggccca 4621 gccaatggcc tgtatggcag cctgggagag ccaacccagg ctctgggcag accttctggc 4681 gccctgggca acttctctct ggaacacggc ggactggccg gacccgagca gtatagagaa 4741 ctgaggcccg gcagccctta cggcgtgcca ccttctccag ctgcccctca gtctctgcca 4801 ggacctcagc ctctgctgtc cagcctggtg taccccgata caagcctggg cctggtgcca 4861 tctggcgctc caggcggacc tcctccaatg agagtgctgg ccggcaatgg ccctagcagc 4921 gatctgagca caggcagctc tggcggctac cccgattttc ctgccagccc tgcctcctgg 4981 ctggacgaag tggatcacgc ccagttctga atcgatcgcc caccccaact tgtttattgc 5041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 5101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 5161 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5341 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5461 gctgcggcga gcggtatcag ctcactcaaa ggacggccgc acatgtaagg aaaattttag 5521 ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa aaaatctaac ctcaagtcaa 5581 ggcttttcta tggaataagg aatggacagc agggggctgt ttcatatact gatgacctct 5641 ttatagccaa cctttgttca tggcagccag catatgggca tatgttgcca aactctaaac 5701 caaatactca ttctgatgtt ttaaatgatt tgccctccca tatgtccttc cgagtgagag 5761 acacaaaaaa ttccaacaca ctattgcaat gaaaataaat ttcctttatt agccagaagt 5821 cagatgctca aggggcttca tgatgtcccc ataatttttg gcagagggaa aaagatctca 5881 gtggtatttg tgagccaggg cattggccac accagccacc accttctgat aggcagcctg 5941 cacctgagga gtgaattcac gcgtttagcc tggcagcata tcgagatcga agtcatcgag 6001 agcgtcagca gggagcatat ccagatcaaa atcatccagg gcatcggcgg gcagcatgtc 6061 caggtcgaag tcgtccaggg cgtcggtggg tccgccgctc tcgcacttca gctgcttctc 6121 gaggccgcag atgatcagtt ccaggccgaa caggaaggcg ggctcggcgc cctgtctgtc 6181 gaacagctcg atggcctgct tcagcagggg gggcatgctg tcggtggtgg gtgtctctct 6241 ttcctctttg gcgacctggt gttcctgttc ttccagcacg cagcccaggg tgaagtggcc 6301 cacggcgctc agggcgtaca gggcgttttc caggctgaag ccctgctggc acaggaaggc 6361 cagctggttt tccagtgtct cgtactgctt ctcggtgggt ctggtgccca ggtgcacttt 6421 ggcgccgtcc cggtggctca gcagggcgca tctgtagctc ttggcgttgt tccgcaggaa 6481 gtcctgccag ctctcgcctt ccagagggca gctgtgggtg tggtgccggt ccagcatctc 6541 gatgggcagg gcgtccagca gggcccgctt gttcttcacg tgccagtaca gggtgggctg 6601 ttccacgccc agtttctggg ccagcttccg ggtggtcagg ccctcgatgc ccacgccgtt 6661 cagcagttcc agggcgctgt tgatcacttt gctcttgtcc agccggctca tgtcgacatt 6721 ctttgccaaa atgatgagac agcacaataa ccagcacgtt gcccaggagc tgtaggaaaa 6781 agaagaaggc atgaacatgg ttagcagagg ctctagagcc gccggtcaca cgccagaagc 6841 cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc cccccgcgga cagccccgag 6901 gctggagagg gagaagggga cggcggcgcg gcgacgcacg aaggccctcc ccgcccattt 6961 ccttcctgcc ggcgccgcac cgcttcgccc cgcgcccgct agagggggtg cggcggcgcc 7021 tcccagattt cggctccgca cagatttggg acaaaggaag tccctgcgcc ctctcgcacg 7081 attaccataa aaggcaatgg ctgcggctcg ccgcgcctcg acagccgccg gcgctccggg 7141 ggccgccgcg cccctccccc gagccctccc cggcccgagg cggccccgcc ccgcccggca 7201 cccccacctg ccgccacccc ccgcccggca cggcgagccc cgcgccacgc cccgtacgga 7261 gccccgcacc cgaagccggg ccgtgctcag caactcgggg aggggggtgc agggggggtt 7321 gcagcccgac cgacgcgccc acaccccctg ctcacccccc cacgcacaca ccccgcacgc 7381 agcctttgtt cccctcgcag ccccccccgc accgcggggc accgcccccg gccgcgctcc 7441 cctcgcgcac actgcggagc gcacaaagcc ccgcgccgcg cccgcagcgc tcacagccgc 7501 cgggcagcgc ggagccgcac gcggcgctcc ccacgcacac acacacgcac gcaccccccg 7561 agccgctccc cccgcacaaa gggccctccc ggagcccctc aaggctttca cgcagccaca 7621 gaaaagaaac aagccgtcat taaaccaagc gctaattaca gcccggagga gaagggccgt 7681 cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg 7741 cggcggcgga gcggggcacg gggcgaaggc agcgtcgcag cgactccccg cccgccgcgc 7801 gcttcgcttt ttatagggcc gccgccgccg ccgcctcgcc ataaaaggaa actttcggag 7861 cgcgccgctc tgattggctg ccgccgcacc tctccgcctc gccccgcccc gcccctcgcc 7921 ccgccccgcc ccgcctggcg cgcgcccccc cccccccccc gcccccatcg ctgcacaaaa 7981 taattaaaaa ataaataaat acaaaattgg gggtggggag gggggggaga tggggagagt 8041 gaagcagaac gtggggctca cctcgaccat ggtaatagcg atgactaata cgtagatgta 8101 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt 8161 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa 8221 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt 8281 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc 8341 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa 8401 tgaccccgta attgattact attaataact agtcaataat caatgtcgag acccatagag 8461 cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc 8521 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca 8581 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag 8641 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttat 8701 aacttcgtat aatgtatgct atacgaagtt atggtacctt actcgtccat gccgccggtg 8761 gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag 8821 gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc 8881 ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc 8941 ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag 9001 cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc 9061 accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg 9121 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag 9181 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg 9241 gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc 9301 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc 9361 acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc 9421 tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcgtaggcgc cggtcacagc 9481 ttggatctgt aacggcgcag aacagaaaac gaaacaaaga cgtagagttg agcaagcagg 9541 gtcaggcaaa gcgtggagag ccggctgagt ctaggtaggc tccaagggag cgccggacaa 9601 aggcccggtc tcgacctgag ctttaaactt acctagacgg cggacgcagt tcaggaggca 9661 ccacaggcgg gaggcggcag aacgcgactc aaccggcgtg gatggcggcc tcaggtaggg 9721 cggcgggcgc gtgaaggaga gatgcgagcc cctcgaagct tcagctgtgt tctggcggca 9781 aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca 9841 ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac 9901 cttctctagg caccggttca attgccgacc cctcccccca acttctcggg gactgtgggc 9961 gatgtgcgct ctgcccactg acgggcaccg gagcgatcgc agatcatgca tccatagagc 10021 ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc 10081 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat 10141 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg 10201 ggcaaacaac agatggctgg caactagaag gcacaggttt ttagaagaac tcgtcaagaa 10261 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 10321 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 10381 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 10441 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 10501 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 10561 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 10621 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 10681 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 10741 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 10801 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt 10861 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 10921 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 10981 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcctagggc 11041 cgggattctc ctccacgtca ccgcatgtta gaagacttcc tctgccctct ccgctgccag 11101 atctctcgag gccctgtggg aggaagagaa gaggtcagat aacttcgtat aatgtatgct 11161 atacgaagtt atgcggccgc tgggagaagg gtaagagagc tcaatcctca tcttacatag 11221 aaggaagtca gtagataata tcttacattt aaaagtcaag aaataacagt ctgaccatac 11281 tatctagaaa tatggaagtt aacagtaagg aagttaaaac agttgaaagt agtttcgtct 11341 agaaagaaag acttggggaa gaacaaagaa ggctaatttt cattatgagc tttattgtaa 11401 cagaaaagag accactaact cagctaggtt ttcccaattc ctcaacactt tggggcacag 11461 tgtttagtca gagctgtgat cactctctga gcacatggat tgtatcttaa ttttcatctc 11521 attgaaaata tgtaccattt gtccaagtgg aaactgggct gaagcatgac tgtgaccacc 11581 accaaagaaa tggctagact cgctaaggtc tgctggggtg acccgtgatt gtgttgtgct 11641 tgatcttgtt gagcatggct cttttaagtc accatacctg cccctgccaa tgcaccaaac 11701 accaacttaa agggactttc agcttttgta agtcagtctc agccttactt ctgaaccatt 11761 tctttgactt ctttcatgac tctgatcctg tcccagtttt aagcctccca gccaccagtc 11821 atgctgtttt gctctgctat ttacattctt ctctacctac tttggacttg atggtatgcc 11881 tactatatta gttatttatt gctgtgtaac aaaccactct gaaacatagt ggcttacaac 11941 cacaaccatt ttattttatt atgaaactgt gggtcagaat gtatattggc ccattggcat 12001 gattctcctg ctattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 12061 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 12121 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 12181 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 12241 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 12301 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 12361 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 12421 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 12481 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 12541 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 12601 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 12661 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 12721 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 12781 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 12841 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 12901 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 12961 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 13021 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 13081 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 13141 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 13201 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 13261 ctacatacct cgctctgcta atcc //