LOCUS Exported 11625 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses S. pyogenes CRISPR chimeric RNA element with customizable sgRNA from U6 promoter and hygromycin resistance marker with 2A iRFP670 from EF-1a promoter. 3rd generation lentiviral backbone.. ACCESSION . VERSION . KEYWORDS lentiGuide-Hygro-iRFP670 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11625) AUTHORS Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I, Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ TITLE Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes. JOURNAL Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27. PUBMED 28757163 REFERENCE 2 (bases 1 to 11625) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11625 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 30..1208 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 261..1199 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1156..1176 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 1215..1224 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1230..2246 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK" CDS 2271..2327 /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" misc_feature 3278..3866 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3331..3351) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3749..3760) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3938..4171 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4249..4370 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4286..4305) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4340..4359 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4410..4545 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4472..4491 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4565..4584) /label=T7 /note="T7 promoter, forward primer" promoter complement(4566..4584) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4594..4611) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4594..4610) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4603..4625) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4752..5207 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4839..4858) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5049..5070 /label=F1ori-F /note="F1 origin, forward primer" promoter 5233..5337 /gene="bla" /label=AmpR promoter CDS 5338..6198 /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(5556..5575) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6369..6957 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6858..6877 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7111..7128 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7245..7266 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7281..7311 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7319..7335 /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)." primer_bind 7324..7346 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7343..7359 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7343..7359 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7378..7398 /label=T3 /note="T3 promoter, forward primer" promoter 7380..7398 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7426..7652 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 7653..7833 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7880..8005 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8498..8731 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8916..8960 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 9138..9378 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 9138..9158 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 9309..9328 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 11268..11343 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 11399..11516 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca 61 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga 121 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 181 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 241 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta 301 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg 361 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc 421 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct 481 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt 541 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc 601 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc 661 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt 721 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct 781 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg 841 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt 901 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg 961 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt 1021 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg 1081 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg 1141 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg 1201 tgtcgtgacg tacggccacc atggaattca aaaagcctga actcaccgct acctctgtcg 1261 agaagtttct gatcgaaaag tttgacagtg tatccgacct gatgcagctc tccgagggcg 1321 aagaatctcg ggctttcagc ttcgatgtgg gagggcgtgg atatgtcctg cgggtgaata 1381 gctgcgccga tggtttctac aaagatcgct atgtttatcg gcactttgca tccgccgctc 1441 tccctattcc cgaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct 1501 cccgccgtgc acagggtgtc accttgcaag acctgcctga aaccgaactg cccgctgttc 1561 tccagcccgt cgccgaggcc atggatgcca tcgctgccgc cgatcttagc cagaccagcg 1621 ggttcggccc attcggacct caaggaatcg gtcaatacac tacatggcgc gatttcatct 1681 gcgctattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg 1741 cctccgtcgc ccaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc 1801 ggcacctcgt gcacgccgat ttcggctcca acaatgtcct gaccgacaat ggccgcataa 1861 cagccgtcat tgactggagc gaggccatgt tcggggattc ccaatacgag gtcgccaaca 1921 tcttcttctg gaggccctgg ttggcttgta tggagcagca gacccgctac ttcgagcgga 1981 ggcatcccga gcttgcagga tctcctcggc tccgggctta tatgctccgc attggtcttg 2041 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gctcagggtc 2101 gctgcgacgc aatcgtccgg tccggagccg ggactgtcgg gcgtacacaa atcgcccgca 2161 gaagcgctgc cgtctggacc gatggctgtg tggaagtgct cgccgatagt ggaaacagac 2221 gccccagcac tcgtcctagg gcaaaggatc tgcagtctag aggatctggt gccacgaatt 2281 tctcactact caagcaggcc ggtgatgtcg aggaaaaccc tggtcctatg gcccggaagg 2341 tggacctgac cagctgcgat agagagccaa tccacatccc cggctccatc cagccatgcg 2401 gatgtctgct ggcatgtgac gcacaggccg tgcggatcac cagaatcaca gagaacgcag 2461 gagccttctt tggaagggag accccaaggg tgggagagct gctggccgat tacttcggag 2521 agacagaggc acacgcactg aggaatgcac tggcacagag ctccgacccc aagagacctg 2581 ccctgatctt cggatggagg gatggactga ccggaagaac atttgacatc tccctgcacc 2641 ggcacgatgg cacatctatc atcgagtttg agccagcagc agcagagcag gcagacaacc 2701 cactgaggct gacccgccag atcatcgcca gaacaaagga gctgaagagc ctggaggaga 2761 tggcagcaag ggtgcctaga tacctgcagg ccatgctggg ctaccacaga gtgatgctgt 2821 atagattcgc cgacgatggc tctggcatgg tcatcggcga ggccaagagg agcgacctgg 2881 agtccttcct gggacagcac tttccagcat ccctggtgcc tcagcaggca aggctgctgt 2941 atctgaagaa cgccatccgc gtggtgtccg attctagggg catctctagc cgcatcgtgc 3001 cagagcacga cgcatctgga gccgccctgg atctgagctt cgcccacctg aggagcatct 3061 ccccttgcca cctggagttt ctgcgcaata tgggcgtgtc tgccagcatg tccctgtcta 3121 tcatcatcga cggcaccctg tggggcctga tcatctgtca ccactatgag cctagggcag 3181 tgccaatggc acagagggtg gcagcagaga tgttcgccga ttttctgtcc ctgcacttta 3241 cagccgccca ccaccagcgg tgacgcgtta agtcgacaat caacctctgg attacaaaat 3301 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 3361 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 3421 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 3481 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 3541 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 3601 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 3661 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 3721 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 3781 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 3841 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct 3901 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa 3961 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc 4021 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga 4081 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga 4141 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt 4201 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt 4261 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 4321 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 4381 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 4441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 4501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa 4561 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga 4621 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 4681 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 4741 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 4801 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 4861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 4921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 4981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 5041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 5101 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 5161 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt 5221 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 5281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 5521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 5581 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 5881 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 6181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 6241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 6301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6481 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 6541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 6721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 6781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 6841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 6901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 7021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 7081 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 7141 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 7201 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 7261 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 7321 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca 7381 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa 7441 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga 7501 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa 7561 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata 7621 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag 7681 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 7741 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 7801 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc 7861 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc 7921 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa 7981 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga 8041 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc 8101 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg 8161 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc 8221 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac 8281 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca 8341 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat 8401 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga 8461 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct 8521 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac 8581 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac 8641 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg 8701 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca 8761 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt 8821 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac 8881 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat 8941 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa 9001 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt 9061 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa 9121 ccccgagggg acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat 9181 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac 9241 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt 9301 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat 9361 atatcttgtg gaaaggacga aacaccggag acggttgtaa atgagcacac aaaatacaca 9421 tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct ttttaataac 9481 tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat aaaaatgagc 9541 aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt aataatagct 9601 actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg agcatcttca 9661 aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac tgacttcatt 9721 agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt tttaacaaat 9781 tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt tgttaataaa 9841 caattttcca taatacattt aacaacatgt gatccagctg ctttttttac agctttcatg 9901 tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa taccatatca 9961 atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt agctgttgtg 10021 catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc ttttaataat 10081 tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc aaattcaatt 10141 gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa taatgtgtga 10201 tctatatatt tgtttagttt cattttttct cctatatatt catttttaat tttaattctt 10261 taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca cctataaaat 10321 aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac ttatcttcta 10381 aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg tagtcttcaa 10441 cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat gatttcatta 10501 atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa gtgaaagtgt 10561 aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt ttattttgtt 10621 ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca tctttatctc 10681 tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca aaatttaatc 10741 cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt aaagttttta 10801 caatattaac tccatatttt tcatgagcga ttctatcacc caaatcactt gttacaaaac 10861 ttgaatatag agccggattt tttggaatgc tatttaagcg ttttagattt gataattttc 10921 aatcaattaa aattggtcct gtttgatttc catctaatct tacaaaatga ccatcatgtt 10981 ttattgccat tccaaatctg tcagcatctg ggtcattcat aataataata tctgcatcat 11041 gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga tttggattta 11101 caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata acgttatatc 11161 ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact gcgctaaaaa 11221 taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt ttagagctag 11281 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg 11341 tgctttttta agcttggcgt aactagatct tgagacaaat ggcagtattc atccacaatt 11401 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag 11461 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg 11521 tttattacag ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa 11581 gatggataaa gttttaaaca gagaggaatc tttgcagcta atgga //