LOCUS Exported 11394 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 dTomato from EF-1a promoter. 3rd generation lentiviral backbone.. ACCESSION . VERSION . KEYWORDS lentiGuide-Hygro-dTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11394) 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 11394) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11394 /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" CDS 2328..3029 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLYGMDELYK" primer_bind complement(2461..2479) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2729..2748 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2923..2946 /label=DsRed1-C /note="DsRed1, forward primer" misc_feature 3047..3635 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3100..3120) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3518..3529) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3707..3940 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4018..4139 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4055..4074) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4109..4128 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4179..4314 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4241..4260 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4334..4353) /label=T7 /note="T7 promoter, forward primer" promoter complement(4335..4353) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4363..4380) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4363..4379) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4372..4394) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4521..4976 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4608..4627) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4818..4839 /label=F1ori-F /note="F1 origin, forward primer" promoter 5002..5106 /gene="bla" /label=AmpR promoter CDS 5107..5967 /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(5325..5344) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6138..6726 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6627..6646 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6880..6897 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7014..7035 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7050..7080 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7088..7104 /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 7093..7115 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7112..7128 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7112..7128 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7147..7167 /label=T3 /note="T3 promoter, forward primer" promoter 7149..7167 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7195..7421 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 7422..7602 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7649..7774 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8267..8500 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8685..8729 /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 8907..9147 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 8907..8927 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 9078..9097 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 11037..11112 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 11168..11285 /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 gtgagcaagg 2341 gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg 2401 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg 2461 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc 2521 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga 2581 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc 2641 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga 2701 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg 2761 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg 2821 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca 2881 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc 2941 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt 3001 tcctgtacgg catggacgag ctgtacaagt gacgcgttaa gtcgacaatc aacctctgga 3061 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 3121 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 3181 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 3241 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 3301 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 3361 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 3421 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac 3481 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 3541 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 3601 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac 3661 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 3721 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 3781 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 3841 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 3901 tccctcagac ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt 3961 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac 4021 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4081 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 4141 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca 4201 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4261 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga 4321 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca 4381 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc 4441 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg 4501 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 4561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 4621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 4681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 4741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 4801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 4861 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 4921 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt 4981 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 5041 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 5101 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 5161 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 5221 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 5281 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 5341 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 5401 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 5461 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 5521 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 5581 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 5641 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 5701 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 5761 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 5821 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 5881 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 5941 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 6001 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 6061 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 6121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 6181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 6241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 6301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 6361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 6421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 6481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 6541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 6601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 6661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 6721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 6781 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 6841 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 6901 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 6961 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 7021 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 7081 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc 7141 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt 7201 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac 7261 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct 7321 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt 7381 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc 7441 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 7501 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 7561 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga 7621 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc 7681 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg 7741 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc 7801 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat 7861 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc 7921 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga 7981 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat 8041 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac 8101 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga 8161 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca 8221 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc 8281 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac 8341 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg 8401 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa 8461 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg 8521 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg 8581 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa 8641 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat 8701 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt 8761 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg 8821 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc 8881 acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc 8941 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga 9001 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa 9061 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct 9121 tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa tgagcacaca 9181 aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca aaatcttctt 9241 tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa acttttaata 9301 aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc ttcaccttta 9361 ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat cattttttga 9421 gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc tttaacaact 9481 gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt 9541 ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat ttcttctttt 9601 gttaataaac aattttccat aatacattta acaacatgtg atccagctgc tttttttaca 9661 gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc aatatttaat 9721 accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa tgcttttgta 9781 gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac atttgtgcct 9841 tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca 9901 aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc ttgttttaat 9961 aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc atttttaatt 10021 ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat tcaccaacac 10081 ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca aatcatgact 10141 tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt ccatcgatgt 10201 agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt gctctcaatg 10261 atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc aagtttaaag 10321 tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct actaatgttt 10381 tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct tgtattccat 10441 ctttatctct agctgagtca tcaattacat atccataact ttcttcataa gcaaaaacaa 10501 aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta aatccagtta 10561 aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc aaatcacttg 10621 ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt tttagatttg 10681 ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt acaaaatgac 10741 catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata ataataatat 10801 ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca aattctggat 10861 ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca acctcaataa 10921 cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt ccattaactg 10981 cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac gtctctgttt 11041 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca 11101 ccgagtcggt gcttttttaa gcttggcgta actagatctt gagacaaatg gcagtattca 11161 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag 11221 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa 11281 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gagggggccc 11341 gggtgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tgga //