LOCUS       Exported                8988 bp DNA     circular SYN 09-JUN-2025
DEFINITION  Vector for tandem expression of two sgRNAs from two independent U6 
            promoters. Cas9 is expressed by Cbh promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8988)
  AUTHORS   Maddalo D, Manchado E, Concepcion CP, Bonetti C, Vidigal JA, Han YC,
            Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura
            A
  TITLE     In vivo engineering of oncogenic chromosomal rearrangements with the
            CRISPR/Cas9 system.
  JOURNAL   Nature. 2014 Dec 18;516(7531):423-7. doi: 10.1038/nature13902. Epub 
            2014 Oct 22.
  PUBMED    25337876
REFERENCE   2  (bases 1 to 8988)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8988)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8988)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature13902"; journalName: "Nature"; date: "2014-12-18-
            18"; volume: "516"; issue: "7531"; pages: "423-7"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        337..412
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        503..743
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        778..853
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        943..1228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1230..1505
                     /label=chicken beta-actin promoter
     intron          1506..1733
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1745..1754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1754..1819
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1826..1846
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1871..5971
                     /codon_start=1
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /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             5972..6019
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    6053..6260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6269..6409
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      6484..6939
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6956..6975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     7075..7097
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7135..7153)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        7221..7325
                     /label=AmpR promoter
     CDS             7326..8186
                     /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"
     CDS             7326..8183
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8357..8945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggccttttgc tcacatgcgc cattctcgag catgcgccat tctcgagcat gctcgagaat
       61 ggcgcatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
      121 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
      181 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
      241 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
      301 tggaaaggac gaaacaccgg gtcttcgaga agacctgttt tagagctaga aatagcaagt
      361 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt
      421 tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat
      481 tctgcagaca aatggctcta gagagggcct atttcccatg attccttcat atttgcatat
      541 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
      601 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
      661 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
      721 tttatatatc ttgtggaaag gacgaaacac cggagaccta ttcgccttaa ggtctcggtt
      781 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      841 accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc
      901 gtttttagcg cgtgcgccaa ttctgcagac aaatggggta cccgttacat aacttacggt
      961 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc
     1021 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1081 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     1141 gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     1201 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     1261 ccccatctcc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     1321 gcagcgatgg gggcgggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
     1381 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
     1441 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
     1501 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
     1561 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
     1621 ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg
     1681 gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac
     1741 cggtgccacc atggactata aggaccacga cggagactac aaggatcatg atattgatta
     1801 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
     1861 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
     1921 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     1981 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     2041 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     2101 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     2161 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     2221 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     2281 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     2341 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     2401 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     2461 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     2521 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     2581 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     2641 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     2701 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     2761 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     2821 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     2881 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     2941 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     3001 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     3061 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     3121 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     3181 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     3241 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     3301 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     3361 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     3421 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     3481 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     3541 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     3601 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     3661 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     3721 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     3781 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     3841 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     3901 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     3961 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     4021 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     4081 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     4141 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     4201 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     4261 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     4321 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     4381 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     4441 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     4501 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     4561 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     4621 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     4681 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     4741 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     4801 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     4861 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     4921 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     4981 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     5041 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     5101 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     5161 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     5221 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     5281 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     5341 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     5401 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     5461 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     5521 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     5581 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     5641 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     5701 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     5761 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     5821 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     5881 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     5941 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
     6001 ccaggcaaaa aagaaaaagt aagaattcct agagctcgct gatcagcctc gactgtgcct
     6061 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6121 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6181 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     6241 aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc
     6301 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     6361 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     6421 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     6481 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6541 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6661 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     6721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6781 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     6841 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     6901 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     6961 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     7021 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7081 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     7141 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     7201 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7261 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7321 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7381 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7441 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7501 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7561 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7621 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7681 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7741 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7801 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7861 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7921 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7981 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8041 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8101 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8161 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8221 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8281 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8341 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8401 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8461 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8521 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8581 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8641 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8701 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8761 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8821 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8881 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8941 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgct
//
