LOCUS Exported 8845 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR-Sirius plasmid. ACCESSION . VERSION . KEYWORDS pPUR-hU6-sgRNA-Sirius-8XMS2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8845) AUTHORS Ma H, Tu LC, Naseri A, Chung YC, Grunwald D, Zhang S, Pederson T TITLE CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging. JOURNAL Nat Methods. 2018 Nov;15(11):928-931. doi: 10.1038/s41592-018-0174-0. Epub 2018 Oct 30. PUBMED 30377374 REFERENCE 2 (bases 1 to 8845) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8845 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 426..444 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 522..1118 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(522..541) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1018..1038 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 1128..1184 /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 1200..1910 /codon_start=1 /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /label=mTagBFP2 /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGIYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" CDS 1911..1931 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" LTR 2027..2260 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2332..2453 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2369..2388) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2423..2442 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2493..2628 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2555..2574 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2648..2667) /label=T7 /note="T7 promoter, forward primer" promoter complement(2649..2667) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2677..2694) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2677..2693) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2686..2708) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2835..3290 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2922..2941) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3132..3153 /label=F1ori-F /note="F1 origin, forward primer" promoter 3316..3420 /gene="bla" /label=AmpR promoter CDS 3421..4281 /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(3639..3658) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4452..5040 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4941..4960 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5194..5211 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5328..5349 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5364..5394 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5402..5418 /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 5407..5429 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5426..5442 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5426..5442 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5461..5481 /label=T3 /note="T3 promoter, forward primer" promoter 5463..5481 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5509..5735 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5736..5916 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5963..6088 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6581..6814 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6999..7043 /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 7341..7581 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 7341..7361 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 7512..7531 /label=LKO.1 5' /note="Human U6 promoter, forward primer" CDS 7936..8241 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 8120..8139 /label=ccdB-fwd /note="ccdB gene, forward primer" misc_feature 8676..8793 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 8842..507 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" ORIGIN 1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg 61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt 121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct 181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa 241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc 301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc 361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc 421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt 481 gaccgaatca ccgacctctc tccccagggg gatccgccac catgaccgag tacaagccca 541 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 601 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 661 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 721 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 781 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 841 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 901 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 961 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 1021 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 1081 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgc tagcggtgct actaacttca 1141 gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggtagt ggaacgcgta 1201 tggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg 1261 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg 1321 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca 1381 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc 1441 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg 1501 aagatggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct 1561 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa 1621 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca 1681 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac gccaagacca 1741 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcatctac tatgtggact 1801 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg 1861 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat ccaaagaaaa 1921 agcggaaagt gtgagtcgac ctcgaggggg ggcccggtac ctttaagacc aatgacttac 1981 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 2041 cactcccaac gaagaaaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 2101 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 2161 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 2221 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 2281 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt 2341 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 2401 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 2461 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2521 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 2581 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc 2641 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg 2701 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 2761 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 2821 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 3001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 3061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 3121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 3181 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 3241 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact 3301 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 3361 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 3421 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 3481 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 3541 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 3601 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 3661 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 3721 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 3781 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 3841 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 3901 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 3961 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 4021 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 4081 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 4141 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 4201 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 4261 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 4321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 4381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 4441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 4501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 4561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 4621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 4681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 4741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 4801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 4861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 4921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 4981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 5041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 5101 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 5161 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 5221 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 5281 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 5341 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 5401 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc 5461 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg 5521 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag 5581 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag 5641 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag 5701 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct 5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 5941 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 6001 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 6061 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 6121 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 6181 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 6241 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 6301 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 6361 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 6421 gcaagcggcc actgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 6481 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 6541 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 6601 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 6661 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 6721 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 6781 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 6841 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 6901 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 6961 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 7021 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 7081 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 7141 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 7201 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 7261 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc 7321 gagcggccgc ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac 7381 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag 7441 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat 7501 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt 7561 tatatatctt gtggaaagga cgaaacaccg gtgtcttcga atgcatggct tactaaaagc 7621 cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat 7681 gtatacccga agtatgtcaa aaagaggtgt gctatgaagc agcgtattac agtgacagtt 7741 gacagcgaca gctatcagtt gctcaaggca tatatgatgt caatatctcc ggtctggtaa 7801 gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc 7861 aggaagggat ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gctgacgaga 7921 acagggactg gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt 7981 ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc 8041 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat 8101 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt 8161 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac 8221 ctgatgttct ggggaatata aaccctgcag gacgaagacc cgtttgagag ctactgccat 8281 gaggaatgac caccaggcat tccgatccga cgatggacca tcaggccatc gagctgcaga 8341 agtgacgacc acgcacttcg gagtgacaga ggaggatcac ccctctggcc accagagtag 8401 agcatcagcc tactccggac aactacggag gaccaccccg taggagtaga gcgaggagca 8461 ccagccctcg cgtgtgacga tgacgatcac gcatcgtcac acaaactact caaatgtccg 8521 aaaggtggca aacactccaa agcagctaaa cggatcaaac atggcagtag caagttcaaa 8581 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgaattc 8641 tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg 8701 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa 8761 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat 8821 ccactttggc cgcggctcga ggggg //