LOCUS Exported 10736 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian (inducible) protein expression, bicistronic expression of cytosolic EmGFP. ACCESSION . VERSION . KEYWORDS pHR-CMV-TetO2_3C-Twin-Strep_IRES-EmGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10736) AUTHORS Elegheert J, Behiels E, Bishop B, Scott S, Woolley RE, Griffiths SC, Byrne EFX, Chang VT, Stuart DI, Jones EY, Siebold C, Aricescu AR TITLE Lentiviral transduction of mammalian cells for fast, scalable and high-level production of soluble and membrane proteins. JOURNAL Nat Protoc. 2018 Nov 19. pii: 10.1038/s41596-018-0075-9. doi: 10.1038/s41596-018-0075-9. PUBMED 30455477 REFERENCE 2 (bases 1 to 10736) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10736 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(21..39) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(22..39) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(329..348) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(562..581) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 681..703 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(741..759) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 827..931 /gene="bla" /label=AmpR promoter CDS 932..1792 /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(1150..1169) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1963..2551 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2452..2471 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2705..2722 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(2792..2812) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2797..3126 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2977..3112 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3039..3058 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 4260..4325 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 4455..4475 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 4900..5034 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4937..4956) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4991..5010 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 5202..5835 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 5882..6007 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6504..6737 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6922..6966 /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 7251..7368 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 7489..7868 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7869..8072 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 8023..8043 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 8074..8092 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 8095..8113 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" regulatory 8177..8186 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 8294..8317 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 8330..8419 /codon_start=1 /product="two Strep-Tag(R)II moieties connected by a linker for enhanced binding to Strep-Tactin(R), an engineered form of streptavidin (Schmidt et al., 2013)" /label=Twin-Strep-tag /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK" misc_feature 8434..9020 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(8601..8618) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8828..8847 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 9021..9740 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(9066..9087) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(9327..9346) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 9674..9695 /label=EGFP-C /note="EGFP, forward primer" misc_feature 9801..10389 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(9854..9874) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(10272..10283) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 10499..10732 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" ORIGIN 1 ctagaattaa ttccgtgtat tctatagtgt cacctaaatc gtatgtgtat gatacataag 61 gttatgtatt aattgtagcc gcgttctaac gacaatatgt acaagcctaa ttgtgtagca 121 tctggcttac tgaagcagac cctatcatct ctctcgtaaa ctgccgtcag agtcggtttg 181 gttggacgaa ccttctgagt ttctggtaac gccgtcccgc acccggaaat ggtcagcgaa 241 ccaatcagca gggtcatcgc tagccagatc ctctacgccg gacgcatcgt ggccggcatc 301 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat 361 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc 421 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg 481 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag 541 cgtcgaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg 601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta 661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc 721 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat 781 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat 841 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata 901 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct 961 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 1021 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa 1081 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt 1141 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg 1201 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca 1261 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa 1321 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt 1381 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 1441 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa 1501 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga 1561 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc 1621 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga 1681 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga 1741 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga 1801 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 1861 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 1921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 1981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 2041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 2101 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 2161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 2221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 2281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 2341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 2401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 2461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 2521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 2581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 2641 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 2701 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc 2761 cgcgcgttgg ccgattcatt aatgcagctg tggaatgtgt gtcagttagg gtgtggaaag 2821 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2881 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2941 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 3001 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 3061 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 3121 tgcaaaaagc ttggacacaa gacaggcttg cgagatatgt ttgagaatac cactttatcc 3181 cgcgtcaggg agaggcagtg cgtaaaaaga cgcggactca tgtgaaatac tggtttttag 3241 tgcgccagat ctctataatc tcgcgcaacc tattttcccc tcgaacactt tttaagccgt 3301 agataaacag gctgggacac ttcacatgag cgaaaaatac atcgtcacct gggacatgtt 3361 gcagatccat gcacgtaaac tcgcaagccg actgatgcct tctgaacaat ggaaaggcat 3421 tattgccgta agccgtggcg gtctgtaccg ggtgcgttac tggcgcgtga actgggtatt 3481 cgtcatgtcg ataccgtttg tatttccagc tacgatcacg acaaccagcg cgagcttaaa 3541 gtgctgaaac gcgcagaagg cgatggcgaa ggcttcatcg ttattgatga cctggtggat 3601 tccggtggta ctgcggttgc gattcgtgaa atgtatccaa aagcgcactt tgtcaccatc 3661 ttcgcaaaac cggctggtcg tccgctggtt gatgactatg ttgttgatat cccgcaagat 3721 acctggattg aacagccgtg ggatatgggc gtcgtattcg tcccgccaat ctccggtcgc 3781 taatcttttc aacgcctggc actgccgggc gttgttcttt ttaacttcag gcgggttaca 3841 atagtttcca gtaagtattc tggaggctgc atccatgaca caggcaaacc tgagcgaaac 3901 cctgttcaaa ccccgcttta aacatcctga aacctcgacg ctagtccgcc gctttaatca 3961 cggcgcacaa ccgcctgtgc agtcggccct tgatggtaaa accatccctc actggtatcg 4021 catgattaac cgtctgatgt ggatctggcg cggcattgac ccacgcgaaa tcctcgacgt 4081 ccaggcacgt attgtgatga gcgatgccga acgtaccgac gatgatttat acgatacggt 4141 gattggctac cgtggcggca actggattta tgagtgggcc ccggatcttt gtgaaggaac 4201 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg 4261 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat 4321 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg 4381 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc 4441 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag 4501 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta 4561 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg 4621 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac 4681 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt 4741 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata 4801 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 4861 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 4921 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4981 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg 5041 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc 5101 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa 5161 agaaattgta tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc 5221 ccaaagaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga 5281 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta 5341 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag 5401 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg 5461 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt 5521 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag 5581 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc 5641 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 5701 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 5761 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 5821 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga 5881 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg 5941 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga 6001 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc 6061 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg 6121 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca 6181 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc 6241 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa 6301 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc 6361 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa 6421 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag 6481 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga 6541 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt 6601 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa 6661 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta 6721 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct 6781 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc 6841 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa 6901 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca 6961 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg 7021 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga 7081 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc 7141 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga 7201 ttagtgaacg gatctcgacg gtatcgccaa atggcagtat tcatccacaa ttttaaaaga 7261 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac 7321 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac 7381 agggacagca gagatccagt ttggatcgat aagcttgata tcgaattgct gcagccccga 7441 tagtcgacta catttatatt ggctcatgtc caatatgacc gccatgttga cattgattat 7501 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt 7561 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 7621 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac 7681 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 7741 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc 7801 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 7861 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac 7921 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc 7981 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc 8041 gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga gatctcccta 8101 tcagtgatag agatcgtcgt cgtgctcgtt tagtgaaccg tctgatctca acaagctgtc 8161 tagagaattc aagcttgcca ccatgggtat ccttcccagc cctgggatgc ctgcgctgct 8221 ctccctcgtg agccttctct ccgtgctgct gatgggttgc gtagctgaaa ccggtgcatc 8281 tggaagtggt acccttgagg tgctgtttca gggaccagga ggtagtggat ctgcttggag 8341 ccatccacag ttcgaaaaag gtggaggttc tggcggtgga tcaggtggaa gtgcatggtc 8401 tcaccctcag tttgagaaat aatgactcga gcgcccctct ccctcccccc cccctaacgt 8461 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac 8521 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag 8581 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa 8641 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag 8701 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga 8761 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag 8821 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtaac 8881 ccattgtatg ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag 8941 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg 9001 atgataatat ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 9061 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 9121 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 9181 tgcccgtgcc ctggcccacc ctcgtgacca ccttgaccta cggcgtgcag tgcttcgccc 9241 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 9301 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 9361 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 9421 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca 9481 ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg 9541 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 9601 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 9661 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 9721 tggacgagct gtacaagtaa agcggccgcg actcttgagt cgagctgcag gcatgcaagc 9781 ttgatatcgt acgtatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg 9841 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 9901 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 9961 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 10021 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 10081 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 10141 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat 10201 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 10261 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 10321 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 10381 cctccccgca tcgataccgt cgtccacgag ggaattaatt cgagctccgt acctttaaga 10441 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 10501 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc 10561 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 10621 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 10681 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcaa //