LOCUS Exported 9465 bp ds-DNA circular SYN 31-JAN-2023 DEFINITION Co-expresses MBP-GSK3-beta_S9A-HA-His (human GSK3-beta with S9A mutation as a fusion protein with MBP, HA, and His-tags) and GST_lambda-PPase in E.coli to produce unphosphorylated MBP-GSK3-beta_S9A-HA-His. ACCESSION . VERSION . KEYWORDS pMG071 MBP-GSK3-beta_S9A-HA-His, GST_lambda-PPase SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9465) AUTHORS Gavagan M, Jameson N, Zalatan JG TITLE The Axin scaffold protects the kinase GSK3-beta from cross-pathway inhibition JOURNAL bioRxiv 2022.12.05.519208 REFERENCE 2 (bases 1 to 9465) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 31, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9465 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 3..80 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 81..1163 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(100..119) /label=LacI-R /note="LacI, reverse primer" promoter 1406..1434 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 1442..1458 /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 1447..1469 /label=M13/pUC Reverse /note="In lacZ gene" CDS 1528..2628 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" regulatory 2485..2494 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 2602..2625 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 2677..2688 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 2731..2751 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3569..3580 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS 4015..4041 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 4069..4086 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" RBS 4116..4138 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4161..4811 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP" primer_bind 4772..4794 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 4848..4868 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" primer_bind complement(5558..5575) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5558..5574) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5567..5589) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(5753..5770) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 5923..6009 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6101..6128 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 6147..6238 /gene="bla" /label=AmpR promoter CDS 6239..7099 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6457..6476) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin complement(7141..7654) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7336..7357) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7548..7567 /label=F1ori-R /note="F1 origin, reverse primer" rep_origin 7765..8353 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8254..8273 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8507..8524 /label=L4440 /note="L4440 vector, forward primer" misc_feature 8539..8679 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(8605..8624) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8765..8787 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(8781..8972) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg 901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1501 gcacttcacc aacaaggacc atagattatg aaaactgaag aaggtaaact ggtaatctgg 1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2461 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcaactagt 2701 ggatctggtg gtggtggcgg ttccatgagc gagaatcttt attttcaggg atcctcaggg 2761 cggcccagaa ccaccgcctt tgcggagagc tgcaagccgg tgcagcagcc ttcagctttt 2821 ggcagcatga aagttagcag agacaaggac ggcagcaagg tgacaacagt ggtggcaact 2881 cctgggcagg gtccagacag gccacaagaa gtcagctata cagacactaa agtgattgga 2941 aatggatcat ttggtgtggt atatcaagcc aaactttgtg attcaggaga actggtcgcc 3001 atcaagaaag tattgcagga caagagattt aagaatcgag agctccagat catgagaaag 3061 ctagatcact gtaacatagt ccgattgcgt tatttcttct actccagtgg tgagaagaaa 3121 gatgaggtct atcttaatct ggtgctggac tatgttccgg aaacagtata cagagttgcc 3181 agacactata gtcgagccaa acagacgctc cctgtgattt atgtcaagtt gtatatgtat 3241 cagctgttcc gaagtttagc ctatatccat tcctttggaa tctgccatcg ggatattaaa 3301 ccgcagaacc tcttgttgga tcctgatact gctgtattaa aactctgtga ctttggaagt 3361 gcaaagcagc tggtccgagg agaacccaat gtttcgtata tctgttctcg gtactatagg 3421 gcaccagagt tgatctttgg agccactgat tatacctcta gtatagatgt atggtctgct 3481 ggctgtgtgt tggctgagct gttactagga caaccaatat ttccagggga tagtggtgtg 3541 gatcagttgg tagaaataat caaggtcctg ggaactccaa caagggagca aatcagagaa 3601 atgaacccaa actacacaga atttaaattc cctcaaatta aggcacatcc ttggactaag 3661 gtcttccgac cccgaactcc accggaggca attgcactgt gtagccgtct gctggagtat 3721 acaccaactg cccgactaac accactggaa gcttgtgcac attcattttt tgatgaatta 3781 cgggacccaa atgtcaaact accaaatggg cgagacacac ctgcactctt caacttcacc 3841 actcaagaac tgtcaagtaa tccacctctg gctaccatcc ttattcctcc tcatgctcgg 3901 attcaagcag ctgcttcaac ccccacaaat gccacagcag cgtcagatgc taatactgga 3961 gaccgtggac agaccaataa tgctgcttct gcatcagctt ccaactccac tagttaccca 4021 tacgatgttc cagattacgc aagcttgggt ggtcccaaca gcggccgcca ccatcaccac 4081 caccactagg ggccgcactc gaggctagaa ataattttgt ttaactttaa gaaggagata 4141 tacatatgaa acaagatctg atgtccccta tactaggtta ttggaaaatt aagggccttg 4201 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg 4261 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca 4321 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt 4381 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa 4441 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag 4501 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg 4561 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca 4621 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc 4681 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga 4741 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg 4801 accatcctcc aactagtgga tctggtggtg gtggcggttc catgagcgag aatctttatt 4861 ttcagggatc catgcgctat tacgaaaaaa ttgatggcag caaataccga aatatttggg 4921 tagttggcga tctgcacgga tgctacacga acctgatgaa caaactggat acgattggat 4981 tcgacaacaa aaaagacctg cttatctcgg tgggcgattt ggttgatcgt ggtgcagaga 5041 acgttgaatg cctggaatta atcacattcc cctggttcag agctgtacgt ggaaaccatg 5101 agcaaatgat gattgatggc ttatcagagc gtggaaacgt taatcactgg ctgcttaatg 5161 gcggtggctg gttctttaat ctcgattacg acaaagaaat tctggctaaa gctcttgccc 5221 ataaagcaga tgaacttccg ttaatcatcg aactggtgag caaagataaa aaatatgtta 5281 tctgccacgc cgattatccc tttgacgaat acgagtttgg aaagccagtt gatcatcagc 5341 aggtaatctg gaaccgcgaa cgaatcagca actcacaaaa cgggatcgtg aaagaaatca 5401 aaggcgcgga cacgttcatc tttggtcata cgccagcagt gaaaccactc aagtttgcca 5461 accaaatgta tatcgatacc ggcgcagtgt tctgcggaaa cctaacattg attcaggtac 5521 agggagaagg cgcatagctg cagtcgagaa gcttggcact ggccgtcgtt ttacaacgtc 5581 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 5641 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 5701 tgaatggcga atggcagctt ggctgttttg gcggatgaga taagattttc agcctgatac 5761 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg 5821 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta 5881 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct 5941 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt 6001 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg 6061 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat 6121 ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta 6181 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 6241 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 6301 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 6361 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 6421 agaacgtttc ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 6481 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 6541 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 6601 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 6661 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 6721 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 6781 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 6841 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 6901 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 6961 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 7021 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 7081 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 7141 accccggttg ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa 7201 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 7261 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata 7321 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 7381 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa 7441 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 7501 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 7561 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 7621 cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga agatcctttt 7681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 7741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 7801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 7861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 7921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 7981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 8041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 8101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 8161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 8221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 8281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 8341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 8401 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 8461 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 8521 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 8581 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta 8641 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc 8701 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 8761 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag 8821 ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc ctgttcatcc 8881 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc 8941 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaatg gggatttctg 9001 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat 9061 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg 9121 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt 9181 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc 9241 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt 9301 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat 9361 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc 9421 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg aaatt //