pAc-sgRNA-Cas9

LOCUS       pAc-sgRNA-Cas9               10632 bp ds-DNA     circular SYN 12-NOV-2018
DEFINITION  Expresses sgRNA and Cas9-Puro in Drosophila S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    pAc-sgRNA-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10632)
  AUTHORS   Bassett AR, Tibbit C, Ponting CP, Liu JL
  TITLE     Mutagenesis and homologous recombination in Drosophila cell lines 
            using CRISPR/Cas9.
  JOURNAL   Biol Open. 2013 Dec 10. pii: bio.20137120v1. doi: 
            10.1242/bio.20137120.
  PUBMED    24326186
REFERENCE   2  (bases 1 to 10632)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 12, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10632
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        complement(167..242)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(261..660)
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     CDS             3136..3201
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3208..3228
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3253..7353
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7354..7401
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7417..7470
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7495..8094
                     /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(7495..7514)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7991..8011
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8102..8236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8126..8145)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8180..8199
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8837..9425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8917..8936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9596..10456)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10219..10238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10457..10561)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10629..15
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggcgtatcac gaggcccttt cgtcttcaag aattaattct atattctaaa aacacaaatg
       61 atacttctaa aaaaaaatca tgaatggcat caactctgaa tcaaatcttt gcagatgcac
      121 ctacttctca tttccactgt cacatcattt ttccagatct aaaaaagcac cgactcggtg
      181 ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc tagctctaaa
      241 acggaagagc tcgctcttcc gaagtattga ggaaaacata cctatataaa tgatcaacat
      301 caggaaagag cagttgagaa ttataagaat tggcaaatgg tcttaagaac cctctgctta
      361 agattttcaa aatttccttt agaatcaaag tgtcctattg ttcgtttgtt gaaaacacag
      421 ttcgatttat tgactataat aaattgatag ttttaaatat agaggcacga ctaagagagt
      481 ttggttttgt tttggttagg gaccaaaaaa agtatacata acgaataaaa aaggatttaa
      541 gtgcaaatgg taaaaaagtg tcgagttttt cttgagttga ttgtgctgta atgagactct
      601 gcattcggca tttgactcgg cttttcctac tcgtgccgta tttcaggctg caagtcgaac
      661 agatctcgct gcctgttatg tggcccacaa accaagacac gttttatggc cattaaagct
      721 ggctgatcgt cgccaaacac caaatacata atgaatatgt acacattcga gaaagaagcg
      781 atcaaagaag cgtcttcggg cggagtagga gaatgcggag gagaaggaga acgagctgat
      841 ctagtatctc tccacaatcc aatgccaact gaccaactgg ccatattcgg agcaatttga
      901 agccaatttc catcgcctgg cgatcgctcc attcttggct atatgttttt caccgttacc
      961 cggggccatt ttcaaagact cgtcggcaag ataagattgt gtcactcgct gtctctcttc
     1021 atttgtcgaa gaatgctgag gaatttcgcg atgacgtcgg cgagtatttt gaagaatgag
     1081 aataatttgt atttatacga aaatcagtta gtggaatttt ctacaaaaac atgttatcta
     1141 tagataattt tgttgcaaaa tatgttgact atgacaaaga ttgtatgtat atctttaatg
     1201 tattctcatt ttcttatgta tttataatgg caatgatgat actgatgata ttttaagatg
     1261 atgccagacc aaaaggcttg aatttctgcg tcttttgccg aacgcagtgc atgtgcaatt
     1321 gttgtttttt ggaatattca attttcggac tgtccgcttt gatttcagtt tcttggctta
     1381 ttcaaaaagc aaagtaaagc caaaaaagcg agatggcaat accaaatgcg gcaaaacggt
     1441 agtggaagga aaggggtgcg gggcagcgga aggaagggtg gggcggggcg tggcggggtc
     1501 tgtggctggg cgcgacgtca ccgacgttgg agccactcct ttgaccatgt gtgcgtgtgt
     1561 gtattattcg tgtctcgcca ctcgccggtt gtttttttct ttttatgctg cgctctctct
     1621 agcgccatct cgcttacgca tgctcaacgc accgcatgtt gccgtttcct tttatgcgtc
     1681 attttggctc gaaataggca attatttaaa caaagattag tcaacgaaaa cgctaaaata
     1741 aataagtcta caatatggtt acttattgcc atgtgtgtgc agccaacgat agcaacaaaa
     1801 gcaacaacac aggtggcttt ccctctttca ctttttgttt gcaagccgcg tgcgagcaag
     1861 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggcgaaaaa
     1921 catcaaggcg cctgatacga atgcatttgc aataacaatt gcgatattta atattgttta
     1981 tgaagctgtt tgacttcaaa acacacaaaa aaaaaaataa aacaaattat ttgaaagaga
     2041 attaggaatc ggacgcttat cgttagggta acaacaagaa atgcttactg agtcacagcc
     2101 tctggaaaac tgccgcaagc cagagagaga gagaaaaaga gggagagcag cttagaccgc
     2161 atgtgcttgt gtgtgaggcg tctctctctt cgtctctgtt gcgcaaacgc atagactgca
     2221 ctgaaaaaat cgattaccta ttttttatga atgaatattt gcactattac tattcaaaac
     2281 tattaagata gcaatcacat tcaatagcca aatactatac cacctgagcg atgcaacgaa
     2341 atgatcaatt tgagcaaaaa tgctgcatat ttaggacggc atcattatag aaatgcttct
     2401 tgctgtgtac ttttctctcg tctggcagct gtttcgccgt tattgttaaa accggcttaa
     2461 gttaggtgtg ttttctacga ctagtgaatg ccctactaga agatgtgtgt tgcacaaaat
     2521 gtccctggaa taaccaattt gaagtgcaga tagcagtaaa cgtaagctaa tatgaatatt
     2581 atttaactgt aatgttttaa tatcgctgga cattactaat aaacccacta taaacacatg
     2641 tacatatgta tgttttggca tacaatgagt agttggggaa aaaatgtgta aaagcaccgt
     2701 gaccatcaca gcataaagat aaccagctga agtatcgaat atgagtaacc cccaaattga
     2761 atcacatgcc gcaactgata ggacccatgg aagtacactc ttcatggcga tatacaagac
     2821 acacacaagc acgaacaccc agttgcggag gaaattctcc gtaaatgaaa acccaatcgg
     2881 cgaacaattc atacccatat atggtaaaag ttttgaacgc gacttgagag cggagagcat
     2941 tgcggctgat aaggttttag cgctaagcgg gctttataaa acgggctgcg ggaccagttt
     3001 tcatatcact accgtttgag ttcttgtgct gtgtggatac tcctcccgac acaaagccgc
     3061 tccatcagcc agcagtcgtc taatccagag accccggatc ggggtaccta ctagtccagt
     3121 gtggtggaat tcatggacta taaggaccac gacggagact acaaggatca tgatattgat
     3181 tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga
     3241 gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     3301 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     3361 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     3421 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     3481 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     3541 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     3601 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     3661 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     3721 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     3781 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     3841 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     3901 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     3961 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     4021 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     4081 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     4141 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     4201 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     4261 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     4321 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     4381 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     4441 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     4501 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     4561 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     4621 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     4681 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     4741 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     4801 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     4861 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     4921 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     4981 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     5041 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     5101 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     5161 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     5221 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     5281 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     5341 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     5401 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     5461 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     5521 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     5581 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     5641 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     5701 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     5761 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     5821 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     5881 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     5941 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     6001 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     6061 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     6121 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     6181 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     6241 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     6301 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     6361 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     6421 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     6481 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     6541 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     6601 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     6661 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     6721 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     6781 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     6841 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     6901 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     6961 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     7021 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     7081 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     7141 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     7201 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     7261 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     7321 acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc
     7381 ggccaggcaa aaaagaaaaa gggatcaggc aagcttgagg gcagaggaag tcttctaaca
     7441 tgcggtgacg tggaggagaa tcccggccct gctagcggta gcggcagcgg tagcatgacc
     7501 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     7561 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     7621 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7681 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7741 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7801 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7861 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7921 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     7981 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     8041 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgactcgag
     8101 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     8161 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     8221 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     8281 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     8341 tcagtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     8401 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     8461 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     8521 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     8581 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     8641 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     8701 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     8761 tagatgacga ccatcaggga cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg
     8821 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     8881 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     8941 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9001 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9061 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9121 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     9181 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     9241 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     9301 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     9361 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     9421 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     9481 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     9541 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     9601 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     9661 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     9721 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     9781 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     9841 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     9901 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     9961 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10021 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10081 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10141 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    10201 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    10261 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    10321 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    10381 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    10441 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    10501 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    10561 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
    10621 cctataaaaa ta
//