LOCUS       sequence-364568-        9671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tissue-specific knock-out in zebrafish, Cas9 and GFP driven by a UAS
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9671)
  AUTHORS   Di Donato V, De Santis F, Auer TO, Testa N, Sanchez-Iranzo H, 
            Mercader N, Concordet JP, Del Bene F
  TITLE     2C-Cas9: a versatile tool for clonal analysis of gene function.
  JOURNAL   Genome Res. 2016 Mar 8.
  PUBMED    26957310
REFERENCE   2  (bases 1 to 9671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
            2016 Mar 8."
COMMENT     Sequence Label: pUAS:Cas9T2AGFP;U6:sgRNA1;U6:sgRNA2
FEATURES             Location/Qualifiers
     source          1..9671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    34..128
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     misc_feature    join(575..762,763..774)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        complement(787..805)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(812..829)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(812..828)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(821..843)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      969..1424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1056..1075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1266..1287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1502..1606
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1607..1675,1676..2467)
                     /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(1825..1844)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2638..3226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3127..3146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3380..3397
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3514..3535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3550..3555,3556..3573,3574..3580)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3588..3604
                     /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     3593..3615
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3612..3628
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3612..3628
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3646..3664
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3646..3663
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    join(3715..3726,3727..3889)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4336..4355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4390..4409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4463..5176,5177..5179,5180..5182))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4466..5176,5177..5179,5180..5182))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4508..4529)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4857..4876
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5116..5137
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5183..5236)
                     /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             complement(5252..5272)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5288..9388)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes MGAS15252 Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             5367..5375
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(9395..9415)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      9424..9433
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(9448..9671,1..15))
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     complement(9480..9501)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
ORIGIN
        1 ctatttatac tccggcgctc gctagagtct ccgctcggag gacagtactc cgctcggagg
       61 acagtactcc gctcggagga cagtactccg ctcggaggac agtactccgc tcggaggaca
      121 gtactccgac ctgcaggcat gcaagcttgg atccgtcgac caaaaaaagc accgactcgg
      181 tgccactttt tcaagttgat aacggactag ccttatttaa acttgctatg ctgtttccag
      241 catagctctt aaactgagac cctctctcgg tctctcgagc tgggagtctg gaggacggct
      301 atatatacca gtgacttctg ggtatgtgtt tcatgtgtgg tatatggctt acttacttat
      361 tcaactcatt tacacttaca tgtgtcagta aatcactgtg tagtagtaaa cctgtaaacc
      421 cttaaattct ttaaaatttg aaaaccagca gtttatgtag gtttaccgta gagttttgga
      481 gctgtaagcg tagtcgaccc atgggatcta tttaaattaa actgggcatc agcgcaattc
      541 aattggtttg gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt
      601 tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat
      661 taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa
      721 ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctgggccca
      781 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
      841 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
      901 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
      961 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1021 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1081 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1141 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1201 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1261 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1321 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1381 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     1441 ctccttacgc atctgtgcgg tatttcacac cgcatcaggt ggcacttttc ggggaaatgt
     1501 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1561 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1621 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1681 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1741 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1801 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1861 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1921 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1981 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2041 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2101 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2161 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2221 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2281 tggctggttt attgctgata aatctggagc cggtgagcgt ggatctcgcg gtatcattgc
     2341 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2401 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2461 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2521 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatgcctta
     2581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3301 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3361 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3421 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3481 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     3541 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     3601 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctattta ggtgacacta
     3661 tagaatactc aagctatgca tccaacgcgt tgggagctct cccatatggt cgagcagagg
     3721 tgtaaaaagt actcaaaaat tttactcaag tgaaagtaca agtacttagg gaaaatttta
     3781 ctcaattaaa agtaaaagta tctggctaga atcttacttg agtaaaagta aaaaagtact
     3841 ccattaaaat tgtacttgag tattaaggaa gtaaaagtaa aagcaagaaa gaaaactaga
     3901 gattcttgtt taagcttccg cggtggagct cgaattaatt catcattgaa aaaaagcacc
     3961 gactcggtgc cactttttca agttgataac ggactagcct tatttaaact tgctatgctg
     4021 tttccagcat agctcttaaa cgagacgcat tagatctaat cgtctcgagc tgggagtctg
     4081 gaggacggct atatatacca gtgacttctg ggtatgtgtt tcatgtgtgg tatatggctt
     4141 acttacttat tcaactcatt tacacttaca tgtgtcagta aatcactgtg tagtagtaaa
     4201 cctgtaaacc cttaaattct ttaaaatttg aaaaccagca gtttatgtag gtttaccgta
     4261 gagttttgga gctgtaagcg taggatcccc cactttgtac aaggtttatt gcagcttata
     4321 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4381 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctacgtaa
     4441 tacgactcac tatagttcta gattacttgt acagctcgtc catgccgaga gtgatcccgg
     4501 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     4561 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     4621 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     4681 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     4741 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     4801 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     4861 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     4921 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     4981 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     5041 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     5101 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     5161 gctcctcgcc cttgctcacc attgggccgg gattctcctc cacgtcaccg catgttagaa
     5221 gacttcctct gccctctccg ccgccggacc tcacctttct cttcttctta ggagatctca
     5281 ccggtgagtc gccacccagc tgagacaggt caatccttgt ttcgtacagg ccggtaatac
     5341 tctgatggat cagtgtagcg tccagcacct ccttagtgga ggtgtacctc tttctatcga
     5401 ttgtagtgtc gaagtacttg aaagctgcag gtgcgcccag gtttgtcaga gtgaacagat
     5461 gaatgatatt ttcggcctgc tccctgatgg gcttatctct gtgtttgttg taagcagaca
     5521 ggaccttatc cagatttgcg tcggccagga tcactctttt agagaactcg ctaatctgtt
     5581 caatgatctc gtccagataa tgcttgtgct gctccacaaa cagctgtttc tgttcgttat
     5641 cctctggaga tcctttcagc ttctcgtagt ggcttgccag atacaggaag ttcacgtact
     5701 tagaaggcag ggccagttca ttgcctttct gcagctctcc agcgcttgcc agcatgcgtt
     5761 tcctgccgtt ttccagctca aacagactgt acttgggcag tttaatgatc aggtctttct
     5821 tcacttcctt ataacctttg gcctccagaa aatcgattgg gttcttctcg aaagagcttc
     5881 tctccataat tgtgataccc agcagctcct tgacagattt cagtttcttg cttttgccct
     5941 tctccacctt agcgaccacc aggacggagt atgccacagt gggagagtca aagcctccat
     6001 acttcttagg gtcccagtct ttctttctag cgatcagctt atcagagttg cgttttggca
     6061 gaatactttc ttttgagaaa ccgcctgtct gcacctcagt tttcttgacg atattcactt
     6121 gaggcatgct caggaccttc ctcacggttg caaaatctct gcctttgtcc cacacgattt
     6181 cgcctgtctc tccgtttgtc tcaatcaggg gccttttgcg gatttctcca ttggccaggg
     6241 tgatctctgt cttaaagaaa ttcatgatgt ttgagtaaaa gaaatacttg gcagtagctt
     6301 ttccaatttc ctgctcggac ttggcgatca tttttctgac atcgtacacc ttatagtctc
     6361 cgtacacgaa ttcgctctcc agttttgggt atttcttgat cagtgcggta ccgaccacgg
     6421 cgttcaggta tgcgtcatgg gcatggtgat aattgttaat ctcgcggacc ttataaaact
     6481 ggaaatcttt cctaaagtcg ctgaccagtt tactcttcag ggtaatgacc ttcacttctc
     6541 tgatcagttt gtcattctca tcgtactttg tgttcatgcg actatccaga atctgtgcga
     6601 cgtgtttagt gatctggcgt gtttccacca gctgcctctt gatgaatcca gctttgtcca
     6661 gttctgacag tccaccgcgc tctgcttttg tcaggttatc aaacttcctc tgagtgatca
     6721 gtttggcatt cagcagctgt ctccagtagt tcttcatttt cttgaccact tcctcactag
     6781 ggacattatc tgattttcct ctgtttttgt cggacctggt cagcaccttg ttgtcaatgc
     6841 tgtcatcttt gatgaaactc tgggggacga tatgatccac gtcgtaatca gacagcctgt
     6901 taatatccag ctcctggtcc acgtacatat ctcttccatt ctgcagatag tacagataca
     6961 gcttctcatt ctgcagctgt gtgttttcca ctgggtgctc tttcaggatc tggctaccca
     7021 gctccttaat gccttcctcg attcttttca tcctctctct actgttcttc tgacccttct
     7081 gggttgtctg attttcgcga gccatctcaa tcacgatgtt ttcgggctta tggcccatga
     7141 ctttcaccag ctcatcgacg attttcactg tctgcagaat gcctttcttg atggctgggc
     7201 ttccagccag gtttgcaatc tgctcatgca gggagtggcc ctgtccagac acctgagcct
     7261 tctggatgtc ctctttgaag gtcagactgt catcgtgaat cagctgcata aaattgcggt
     7321 tggcgaagcc atcacttttc agaaagtcca ggattgtctt tcctgactgt ttatccctaa
     7381 tgccgttgat cagcttcctg ctcagtctac cccagccagt gtagcgcctt ctcttcagct
     7441 gtttcatcac cttgtcatcg aacagatgag cataggtttt cagtctttcc tcgatcattc
     7501 ctctgtcctc aaacagggtc agtgtcagca caatgtcttc caggatatcc tcattttcct
     7561 cgttgtccag gaagtcttta tccttaatga tcttcagcag atcgtggtaa gcgcccaggc
     7621 ttgcgttaaa tctgtcttcg actccgctga tctccacgga atcgaagcat tcaattttct
     7681 tgaagtagtc ctccttcagc tgcttgacag tcactttcct gttggtttta aacagcagat
     7741 ccacgattgc tttcttctgc tctcctgaca ggaaggctgg ctttctcatg ccctcggtga
     7801 cgtatttcac ctttgtcagt tcgttataga cagtgaaata ctcgtacagc agggagtgct
     7861 taggcagcac tttctcattg ggcaggtttt tgtcaaaatt agtcattctc tcgatgaagg
     7921 actgtgcaga ggcgccctta tcgaccactt cctcgaagtt ccaaggagta atggtttcct
     7981 cggatttgcg agtcatccag gcaaatctgc tgtttccgcg agccagtgga cccacatagt
     8041 aagggattct gaaggtcaga attttttcga tcttctcgcg gttatctttc agaaaggggt
     8101 agaagtcttc ctggcgcctc aggatagcgt gcagctctcc cagatgaatc tggtgtggga
     8161 tggaaccgtt atcaaatgtg cgctgcttcc tcagcaggtc ctctctattc agtttcacca
     8221 gcagttcctc agttccgtcc atcttctcca gaataggttt gataaacttg tagaattcct
     8281 cctgtgatgc gcctccgtcg atataaccgg cgtagccgtt cttggactga tcaaagaaaa
     8341 tttccttgta cttctctggc agctgctgcc tgaccagagc cttcagcagg gtcagatcct
     8401 gatggtgctc gtcatatctc ttgatcatgc tggcactcag aggagctttt gtaatctcgc
     8461 tgttcactct cagaatatct gacagcagga ttgcgtcgga cagatttttg gcagccagga
     8521 acaggtcagc gtactgatct ccgatctgtg ccagcaggtt atccaggtca tcgtcatatg
     8581 tgtcctttga cagctgcagt ttggcgtcct cagccagatc aaaattgctt ttgaagttag
     8641 gagtcaggcc cagactcaga gcgatcagat ttccaaacag accgttcctc ttttctccgg
     8701 gcagctgtgc aatcaggttc tccagtctgc gtgacttgga cagcctggct gacaggatag
     8761 cttttgcgtc cactctggaa gcattaatag ggttttcctc aaacagctga ttgtaaatct
     8821 ggaccagctg gatgaacagc ttgtccacat cggaattgtc ggggttcaga tcgccctcga
     8881 tcagaaaatg acccctgaac ttaatcatgt gggccagagc cagatagatc agtctcaggt
     8941 cggctttatc agtagagtca gccagtttct tgcgcagatg atagatggtt gggtattttt
     9001 cgtgataggc gacctcgtcc acaatgtttc caaagatagg atggcgctcg tgtttcttat
     9061 cttcctcgac caggaaactt tcctccagcc tatgaaagaa tgaatcgtcc accttagcca
     9121 tttcgttaga gaaaatctcc tgcaggtaac agattctatt cttccttctg gtatagcgcc
     9181 ttcttgctgt tcttttcagg cgtgttgctt cggcagtctc tccagaaccg aacagcaggg
     9241 ctccaatcag atttttcttg atgctgtgcc tgtcggtgtt acccagcacc ttaaatttct
     9301 tgctagggac cttgtagtcg tctgtaatca ctgcccaacc cacggagtta gttccaatat
     9361 ccaggccgat gctatacttc ttatccatgc ttcccacctt tctcttcttc ttaggtggag
     9421 aagccatggt ggccccaatt ccctattcag agttctcttc ttgtattcaa taattacttc
     9481 ttggcagatt tcagtagttg cagttgattt acttggttgc tggttacttt taattgattc
     9541 actttaactt gcactttact gcagattgtt tagcttgttc agctgcgctt gtttatttgc
     9601 ttagctttcg cttagcgacg tgttcacttt gcttgtttga attgaattgt cgctccgtag
     9661 acgaagcgcc t
//
