LOCUS       sequence-224306-        8383 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses SpdCas9-Vp64 fusion protein when packaged into AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8383)
  AUTHORS   Matharu N, Rattanasopha S, Tamura S, Maliskova L, Wang Y, Bernard A,
            Hardin A, Eckalbar WL, Vaisse C, Ahituv N
  TITLE     CRISPR-mediated activation of a promoter or enhancer rescues obesity
            caused by haploinsufficiency.
  JOURNAL   Science. 2018 Dec 13. pii: science.aau0629. doi: 
            10.1126/science.aau0629.
  PUBMED    30545847
REFERENCE   2  (bases 1 to 8383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2018 Dec 13. pii: science.aau0629. doi: 10.1126/science.aau0629."
COMMENT     Sequence Label: pCMV-SpdCas9-VP64
FEATURES             Location/Qualifiers
     source          1..8383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        186..565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        566..769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     720..740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     814..833
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        814..832
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             858..923
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             930..950
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(960..986,987..989,990..3476,3477..3479,3480..5063)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5088..5108
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5118..5267
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5274..5300
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5357..5374)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5363..5587
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5646..5786
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5646..5775
                     /label=AAV2 ITR
     rep_origin      5861..6316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5948..5967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6158..6179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6333..6352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6452..6474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6512..6530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6598..6702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6703..6771,6772..7563)
                     /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(6921..6940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7734..8322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8223..8242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttttgc gctgcttcgc gatgtacggg ccagatatac
      181 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      241 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      301 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      361 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      421 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      481 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      541 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      601 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      661 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      721 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      781 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      841 agctggctag cgccatggac tacaaagacc atgacggtga ttataaagat catgacatcg
      901 attacaagga tgacgatgac aagatggccc ccaagaagaa gaggaaggtg ggccgcggaa
      961 tggataagaa atactcaata ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga
     1021 tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc
     1081 acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag
     1141 cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt
     1201 atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac
     1261 ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa
     1321 atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa
     1381 aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata
     1441 tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg
     1501 tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta
     1561 ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac
     1621 gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc
     1681 tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag
     1741 atgctaaatt acagctttca aaagatactt acgatgatga tttagataat ttattggcgc
     1801 aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt
     1861 tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa
     1921 tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac
     1981 aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag
     2041 gttatattga tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag
     2101 aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca
     2161 agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg
     2221 ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg
     2281 aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc
     2341 gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag
     2401 ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa
     2461 atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt
     2521 ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt
     2581 caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg
     2641 ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt
     2701 caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta
     2761 ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt
     2821 taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc
     2881 acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac
     2941 gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag
     3001 attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata
     3061 gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac
     3121 atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg
     3181 taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta
     3241 ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta
     3301 tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg
     3361 ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag
     3421 acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatgcca
     3481 ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg
     3541 ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa
     3601 actattggag acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa
     3661 cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat
     3721 tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata
     3781 ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta
     3841 aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt
     3901 accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat
     3961 atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa
     4021 tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta
     4081 atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc
     4141 ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg
     4201 ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac
     4261 agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg
     4321 ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt
     4381 attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta
     4441 aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact
     4501 ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat
     4561 atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac
     4621 aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc
     4681 attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc
     4741 agcataagca ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta
     4801 ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac
     4861 caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc
     4921 ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag
     4981 aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg
     5041 atttgagtca gctaggaggt gacccaattg ccggatccaa ggctagcccg aaaaagaaac
     5101 gcaaagttgg gcgcgccgac gcgctggacg atttcgatct cgacatgctg ggttctgatg
     5161 ccctcgatga ctttgacctg gatatgttgg gaagcgacgc attggatgac tttgatctgg
     5221 acatgctcgg ctccgatgct ctggacgatt tcgatctcga tatgttaatt aactacccgt
     5281 acgacgttcc ggactacgct tcttgaatcg gtaggaattc gcggccgtct agacttaagt
     5341 ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5401 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5461 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5521 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     5581 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ggaccgagcg
     5641 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     5701 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     5761 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     5821 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     5881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     5941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6061 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6181 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     6241 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6301 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6361 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6421 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6481 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6541 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6601 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6661 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6721 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6781 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6841 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6901 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6961 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7021 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7081 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7141 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7201 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7261 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7321 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7381 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7441 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7501 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7561 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7621 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7681 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7801 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7861 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7921 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7981 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8041 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8101 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8161 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8221 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8281 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8341 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
