LOCUS Exported 8859 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tet-regulated miR-E (miR-30 variant)-based RNAi. ACCESSION . VERSION . KEYWORDS LT3REV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8859) AUTHORS Fellmann C, Hoffmann T, Sridhar V, Hopfgartner B, Muhar M, Roth M, Lai DY, Barbosa IA, Kwon JS, Guan Y, Sinha N, Zuber J TITLE An optimized microRNA backbone for effective single-copy RNAi. JOURNAL Cell Rep. 2013 Dec 26;5(6):1704-13. doi: 10.1016/j.celrep.2013.11.020. Epub 2013 Dec 12. PUBMED 24332856 REFERENCE 2 (bases 1 to 8859) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8859 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 121..138 /label=L4440 /note="L4440 vector, forward primer" protein_bind 255..276 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 291..321 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 329..345 /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 334..356 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 353..369 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 353..369 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 388..408 /label=T3 /note="T3 promoter, forward primer" promoter 390..408 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 436..662 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 663..843 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 890..1015 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1508..1741 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1926..1970 /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 2237..2354 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2372..2750 /label=TRE3G promoter /note="3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On(R) 3G" protein_bind 2380..2398 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2416..2434 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2452..2470 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2488..2506 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2524..2542 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2560..2578 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2596..2614 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 2659..2683 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2774..3451 /codon_start=1 /product="improved tetrameric variant of DsRed fluorescent protein" /label=DsRed2 /note="mammalian codon-optimized" /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH LFL" primer_bind complement(2954..2974) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 3366..3389 /label=DsRed1-C /note="DsRed1, forward primer" promoter 3715..4214 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3750..3773) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4147..4166 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4237..4956 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4282..4303) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4543..4562) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4890..4911 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5018..5606 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5071..5091) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5489..5500) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5813..6046 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6118..6239 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6155..6174) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6209..6228 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6279..6414 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6341..6360 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6434..6453) /label=T7 /note="T7 promoter, forward primer" promoter complement(6435..6453) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6463..6480) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6463..6479) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6472..6494) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6621..7076 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6708..6727) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6918..6939 /label=F1ori-F /note="F1 origin, forward primer" promoter 7102..7206 /gene="bla" /label=AmpR promoter CDS 7207..8067 /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(7425..7444) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8238..8826 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8727..8746 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 61 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 121 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg 181 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 241 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt 301 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 361 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg 421 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga 481 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag 541 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac 601 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa 661 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 721 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 781 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 841 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg 901 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac 961 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat 1021 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa 1081 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa 1141 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc 1201 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg 1261 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga 1321 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag 1381 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac 1441 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag 1501 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag 1561 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga 1621 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca 1681 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc 1741 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta 1801 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca 1861 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc 1921 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt 1981 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg 2041 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt 2101 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa 2161 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc 2221 gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa 2281 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa 2341 aaattcaaaa ttttatcgat cctctagatt cgagtttact ccctatcagt gatagagaac 2401 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca 2461 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt 2521 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac 2581 gtatatccag tttactccct atcagtgata gagaacgtat aagctttagg cgtgtacggt 2641 gggcgcctat aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga gcaattccac 2701 aacacttttg tcttatacca actttccgta ccacttccta ccctcgtaaa gtcgagcttg 2761 cgttggatcc accatggcct cctccgagaa cgtcatcacc gagttcatgc gcttcaaggt 2821 gcgcatggag ggcaccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg 2881 cccctacgag ggccacaaca ccgtgaagct gaaggtgacc aagggcggcc ccctgccctt 2941 cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc 3001 cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt 3061 gatgaacttc gaggacggcg gcgtggcgac cgtgacccag gactcctccc tgcaggacgg 3121 ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc ccctccgacg gccccgtgat 3181 gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt 3241 gctgaagggc gagacccaca aggccctgaa gctgaaggac ggcggccact acctggtgga 3301 gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg cccggctact actacgtgga 3361 cgccaagctg gacatcacct cccacaacga ggactacacc atcgtggagc agtacgagcg 3421 caccgagggc cgccaccacc tgttcctgta gcggccgcct cgactaggga taacagggta 3481 attgtttgaa tgaggcttca gtactttaca gaatcgttgc ctgcacatct tggaaacact 3541 tgctgggatt acttcgactt cttaacccaa cagaaggctc gagcaaccag aattcgagca 3601 attatcttgt ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct 3661 gaattaaaat ggtataaatt aaatcacttt tttcaattga cgcgtaattc taccgggtag 3721 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt 3781 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac 3841 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt 3901 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca 3961 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca 4021 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt 4081 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag 4141 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat 4201 ctccgggcct ttcgacctgc agcccaagct tccaccatgg tgagcaaggg cgaggagctg 4261 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 4321 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc 4381 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc 4441 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 4501 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 4561 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 4621 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 4681 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc 4741 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc 4801 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg 4861 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 4921 gggatcactc tcggcatgga cgagctgtac aagtaagaat tgttcaattc ctgatcatcc 4981 tgcaggatgc atgtcgactt cgaaacgcgt ctggaacaat caacctctgg attacaaaat 5041 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 5101 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 5161 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 5221 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 5281 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 5341 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 5401 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct 5461 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 5521 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 5581 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa 5641 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa 5701 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg 5761 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg 5821 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta 5881 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa 5941 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac 6001 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt 6061 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac 6121 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 6181 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 6241 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca 6301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 6361 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga 6421 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca 6481 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc 6541 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg 6601 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 6661 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 6721 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 6781 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 6841 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 6901 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 6961 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 7021 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt 7081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 7141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 7201 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 7261 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 7321 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 7381 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 7441 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 7501 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 7561 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 7621 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 7681 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 7741 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 7801 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 7861 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 7921 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 7981 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 8041 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 8101 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 8161 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 8221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 8281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 8341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 8401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 8461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 8521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 8581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 8641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 8701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 8761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 8821 tggaaaaacg ccagcaacgc ggccttttta cggttcctg //