LOCUS Exported 11810 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Express 3xNLS-NGN2 TALE-GCN4 engineered to bind a site in the human NGN2 ene. ACCESSION . VERSION . KEYWORDS EF1a-3NLS-NGN2TALE-GCN4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11810) AUTHORS Zhao C, Zhang Y, Zhao Y, Ying Y, Ai R, Zhang J, Wang Y TITLE Multiple Chemical Inducible Tal Effectors for Genome Editing and Transcription Activation. JOURNAL ACS Chem Biol. 2018 Mar 16;13(3):609-617. doi: 10.1021/acschembio.7b00606. Epub 2018 Jan 8. PUBMED 29308880 REFERENCE 2 (bases 1 to 11810) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11810 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 130..341 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 1205..1214 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1226..1246 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1250..1270 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1274..1294 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4001..4057 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4073..4129 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4145..4201 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4217..4273 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4289..4345 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4361..4417 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4433..4489 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4505..4561 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4577..4633 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4649..4705 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" primer_bind complement(4929..4949) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4934..5263 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5114..5249 /label=SV40 ori /note="SV40 origin of replication" CDS 5273..5647 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" misc_feature 5733..6321 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5786..5806) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6204..6215) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6408..6641 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6713..6834 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 6853..6988 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(7008..7027) /label=T7 /note="T7 promoter, forward primer" promoter complement(7009..7027) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7037..7054) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7037..7053) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7046..7068) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7195..7650 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7282..7301) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7492..7513 /label=F1ori-F /note="F1 origin, forward primer" promoter 7676..7780 /gene="bla" /label=AmpR promoter CDS 7781..8641 /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(7999..8018) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8812..9400 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9301..9320 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9554..9571 /label=L4440 /note="L4440 vector, forward primer" protein_bind 9688..9709 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9724..9754 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9762..9778 /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 9767..9789 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9786..9802 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9786..9802 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9821..9841 /label=T3 /note="T3 promoter, forward primer" promoter 9823..9841 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 9869..10095 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 10096..10276 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 10323..10448 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 10941..11174 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 11359..11403 /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" misc_feature 11670..11787 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 atagcccggg ctcgaccata tgataagctt tgcaaagatg gataaagttt taaacagaga 61 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 121 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 181 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 241 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 301 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 361 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 421 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 481 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 541 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 601 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 661 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 721 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 781 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 841 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 901 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 961 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1021 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1081 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1141 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctcctttc 1201 tagagccacc atgggtggcg cgcctccaaa aaagaagaga aaggtagatc caaaaaagaa 1261 gagaaaggta gatccaaaaa agaagagaaa ggtaggatta attaatggaa gtggaagtgg 1321 aagtggaggt accccggatg ttccagatta cgcttcgcgg acccggctcc cttccccacc 1381 cgcacccagc ccagcgtttt cggccgactc gttctcagac ctgcttaggc agttcgaccc 1441 ctcactgttt aacacatcgt tgttcgactc ccttcctccg tttggggcgc accatacgga 1501 ggcggccacc ggggagtggg atgaggtgca gtcgggattg agagctgcgg atgcaccacc 1561 cccaaccatg cgggtggccg tcaccgctgc ccgaccgccg agggcgaagc ccgcaccaag 1621 gcggagggca gcgcaaccgt ccgacgcaag ccccgcagcg caagtagatt tgagaacttt 1681 gggatattca cagcagcagc aggaaaagat caagcccaaa gtgaggtcga cagtcgcgca 1741 gcatcacgaa gcgctggtgg gtcatgggtt tacacatgcc cacatcgtag ccttgtcgca 1801 gcaccctgca gcccttggca cggtcgccgt caagtaccag gacatgattg cggcgttgcc 1861 ggaagccaca catgaggcga tcgtcggtgt ggggaaacag tggagcggag cccgagcgct 1921 tgaggccctg ttgacggtcg cgggagagct gagagggcct ccccttcagc tggacacggg 1981 ccagttgctg aagatcgcga agcggggagg agtcacggcg gtcgaggcgg tgcacgcgtg 2041 gcgcaatgcg ctcacgggag cacccctcaa cctgacccca gagcaggtcg tggcaattgc 2101 gagccataac gggggaaagc aggcactcga aaccgtccag aggttgctgc ctgtgctgtg 2161 ccaagcgcac ggacttacgc cagagcaggt cgtggcaatt gcgagcaaca tcgggggaaa 2221 gcaggcactc gaaaccgtcc agaggttgct gcctgtgctg tgccaagcgc acggactaac 2281 cccagagcag gtcgtggcaa ttgcgagcaa catcggggga aagcaggcac tcgaaaccgt 2341 ccagaggttg ctgcctgtgc tgtgccaagc gcacgggttg accccagagc aggtcgtggc 2401 aattgcgagc aacggagggg gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt 2461 gctgtgccaa gcgcacggct taaccccaga gcaggtcgtg gcaattgcga gccataacgg 2521 gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg 2581 cctgacccca gagcaggtcg tggcaattgc gagcaacatc gggggaaagc aggcactcga 2641 aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggactgacac cagagcaggt 2701 cgtggcaatt gcgagcaacg gagggggaaa gcaggcactc gaaaccgtcc agaggttgct 2761 gcctgtgctg tgccaagcgc acggactcac tccagagcag gtcgtggcaa ttgcgagcca 2821 taacggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc 2881 gcacggcctc accacagagc aggtcgtggc aattgcgagc aacatcgggg gaaagcaggc 2941 actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggat taaccccaga 3001 gcaggtcgtg gcaattgcga gcaacggagg gggaaagcag gcactcgaaa ccgtccagag 3061 gttgctgcct gtgctgtgcc aagcgcacgg acttacccca gagcaggtcg tggcaattgc 3121 gagcaacatc gggggaaagc aggcactcga aaccgtccag aggttgctgc ctgtgctgtg 3181 ccaagcgcac gggctcaccc cagagcaggt cgtggcaatt gcgagcaaca tcgggggaaa 3241 gcaggcactc gaaaccgtcc agaggttgct gcctgtgctg tgccaagcgc acggtcttac 3301 cccagagcag gtcgtggcaa ttgcgagcaa cggaggggga aagcaggcac tcgaaaccgt 3361 ccagaggttg ctgcctgtgc tgtgccaagc gcacggcctc accccagagc aggtcgtggc 3421 aattgcgagc aacatcgggg gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt 3481 gctgtgccaa gcgcatgggc tcaccccaga gcaggtcgtg gcaattgcga gccatgacgg 3541 gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg 3601 tctaacccca gagcaggtcg tggcaattgc gagccataac gggggaaagc aggcactcga 3661 aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggactcacgc ctgagcaggt 3721 agtggctatt gcatccaata acgggggcag acccgcactg gagtcaatcg tggcccagct 3781 ttcgaggccg gaccccgcgc tggccgcact cactaatgat catcttgtag cgctggcctg 3841 cctcggcgga cgacccgcct tggatgcggt gaagaagggg ctcccgcacg cgcctgcatt 3901 gattaagcgg accaacagaa ggattcccga gaggacatca catcgagtgg cagctagcgg 3961 cggatccgcc atcatacacg cggccgcaac aagcggtccg gaagaacttt tgagcaagaa 4021 ttatcatctt gagaacgaag tggctcgtct taagaaaggt tctggcagtg gagaagaact 4081 gctttcaaag aattaccacc tggaaaatga ggtagctaga ctgaaaaagg ggagcggaag 4141 tggggaggag ttgctgagca aaaattatca tttggagaac gaagtagcac gactaaagaa 4201 agggtccgga tcgggtgagg agttactctc gaaaaattat catctcgaaa acgaagtggc 4261 tcggctaaaa aagggcagtg gttctggaga agagctatta tctaaaaact accacctcga 4321 aaatgaggtg gcacgcttaa aaaagggaag tggcagtggt gaagagctac tatccaagaa 4381 ttatcatctt gagaacgagg tagcgcgttt gaagaagggt tccggctcag gagaggaact 4441 gctctcgaag aactatcatc ttgaaaatga ggtcgctcga ttaaaaaagg gatcgggcag 4501 tggtgaggaa ctactttcaa agaattacca cctcgaaaac gaagtagctc gattaaagaa 4561 aggttcaggg tcgggtgaag aattactgag taaaaattat catctggaaa atgaggtagc 4621 gagactaaaa aaggggagtg gttctggcga ggaattgcta tcgaaaaatt atcatcttga 4681 gaacgaagtt gctaggctca aaaagggctc aggctcaggc tgagtttaaa catcgcctgc 4741 aggggaagtg gaagtgatat cggaagtgga agcactagtg gaagtttcga agtcgacttg 4801 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 4861 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatgtc 4921 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 4981 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 5041 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 5101 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 5161 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 5221 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatggccaa 5281 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg 5341 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg 5401 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct 5461 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc 5521 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg 5581 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca 5641 ggactgaccg atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga 5701 aagaccccac ctgtaggttt ggcaagctcg acaatcaacc tctggattac aaaatttgtg 5761 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 5821 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 5881 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 5941 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 6001 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 6061 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 6121 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg 6181 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 6241 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 6301 ccctttgggc cgcctccccg cctggaattc gagctcagta cctttaagac caatgactta 6361 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6421 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca 6481 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6541 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6601 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6661 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6721 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6781 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6841 ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6901 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6961 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt 7021 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 7081 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag 7141 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc 7201 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 7261 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 7321 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 7381 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 7441 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 7501 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 7561 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 7621 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg 7681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 7741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 7801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 7861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 7921 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 7981 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 8041 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 8101 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 8161 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 8221 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 8281 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac 8341 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 8401 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 8461 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 8521 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 8581 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 8641 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 8701 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 8761 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 8821 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 8881 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 8941 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 9001 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 9061 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 9121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccaa 9181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 9241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 9301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 9361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 9421 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 9481 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 9541 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 9601 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 9661 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca 9721 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt 9781 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag 9841 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc 9901 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc 9961 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg 10021 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta 10081 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc 10141 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 10201 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 10261 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg 10321 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc 10381 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg 10441 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc 10501 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac 10561 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac 10621 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag 10681 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca 10741 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc 10801 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta 10861 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga 10921 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc 10981 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata 11041 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc 11101 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag 11161 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg 11221 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg 11281 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa 11341 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt 11401 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata 11461 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt 11521 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac 11581 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta 11641 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac 11701 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa 11761 aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga //