LOCUS       sequence-209040-       13821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of heat shock inducible Cas9-GFP and U6-driven 5 
            individual gRNAs for scGESTALT in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13821)
  AUTHORS   Raj B, Wagner DE, McKenna A, Pandey S, Klein AM, Shendure J, Gagnon 
            JA, Schier AF
  TITLE     Simultaneous single-cell profiling of lineages and cell types in the
            vertebrate brain.
  JOURNAL   Nat Biotechnol. 2018 Mar 28. pii: nbt.4103. doi: 10.1038/nbt.4103.
  PUBMED    29608178
REFERENCE   2  (bases 1 to 13821)
  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: "Nat 
            Biotechnol. 2018 Mar 28. pii: nbt.4103. doi: 10.1038/nbt.4103."
COMMENT     Sequence Label: pTol2-hsp70l:Cas9-t2A-GFP, 5xU6:sgRNA
FEATURES             Location/Qualifiers
     source          1..13821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(146..167)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(516..527,528..690)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1063..1079
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1099..1119
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    2639..2663
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2665..2681
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2691..2710
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2691..2709
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2745..2754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2763..2783
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2790..6890
                     /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             6803..6811
                     /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             6906..6926
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6942..6995
                     /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             join(7002..7004,7005..7007,7008..7721)
                     /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             join(7002..7004,7005..7007,7008..7718)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7047..7068)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7308..7327)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7655..7676
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(7758..7776)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(7805..7829)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7831..7965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(8028..8048)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(8056..8072)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    10780..10914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(11353..11540,11541..11552)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(11586..11602)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11586..11602)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(11639..11657)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11678..11694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11691..11713)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11702..11718
                     /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)."
     promoter        complement(join(11726..11732,11733..11750,11751..11756))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11771..11792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11909..11926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12080..12668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12160..12179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12839..13630,13631..13699))
                     /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     13462..13481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13700..13804)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
       61 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      121 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
      181 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac gctggctaag
      241 aactcatcag cctccccggt ccatctaccc acgtaccaat gcaccaattg gccacaatga
      301 cggctactac atggtgccat tccttcctct ttataggaat ggagactacc tcctgtccaa
      361 caatgctctt ggatacgagt acgcctacct gttggaccca ggtcattgca caacaccaga
      421 aatgccctct gatctgcaaa agacgtgaat atctgttcag acacccatat ccactctgtt
      481 ccacacaggt cagaggtttg tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa
      541 ttttactcaa gtgaaagtac aagtacttag ggaaaatttt actcaattaa aagtaaaagt
      601 atctggctag aatcttactt gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga
      661 gtattaagga agtaaaagta aaagcaagaa agaaaactag agattcttgt ttaagctttt
      721 aatctcaaaa aacattaaat gaaatgcata caaggtttta tcctgcttta gaactgtttg
      781 tatttaatta tcaaactata agacagacaa tctaatgcca gtacacgcta ctcaaagttg
      841 taaaacctca gatttaactt cagtagaagc tgattctcaa aattgttagt gtcaagccta
      901 gctcttttgg ggctgaaaag caatcctgca gtgctgaaaa gcctctcaca ggcagccgat
      961 gcgggaagag gtgtattagt cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga
     1021 gactccctgg tgtctgaaac acaggccaga tgggccctcg agcaggaaac agctatgacc
     1081 atgattacgc caagctatca actttgtata gaaaagttgc ttcaggggtg tcgcttggtt
     1141 atttccaaaa atcaaattaa ttttattaaa ctattagaac gagcatgttt tgtctatatg
     1201 ctacagaaga taaaaaataa taggagttaa cagttataaa acaacacact ttgtttctat
     1261 tgattgttga ccacactggg gtctcattaa gttagattaa agacacacta actgggtcaa
     1321 aagcagcaga ttgatttcat agcaccaggg taaactttct aacactttta cggcaatcat
     1381 atacattaaa attaaataca gaccacgact gaacaaggag gatgatctcc aatattaaac
     1441 aaagagactt gtgcctattt ctctgagggt aaacatgacc tctcaagtta gcaagttggt
     1501 tttaacacta caaaaatagt taagactgca atcccagaat aaagtattgg ttttaaccaa
     1561 tcaatatagt acagtaaaca tccatttgtt ttgttgaaac gttaaacaaa tctgaccaaa
     1621 gctattagct tatataaaac aggtttgcct tctatgtagc tgaaaacacc acaggcccga
     1681 ttttgctact gtgtaaaaca tttcagcaag atttttttat tgcatttttt tttactgaat
     1741 cgttcaaaca ttttatcatt ttagtttgtt cattcattgc aactggaaaa acaacacatc
     1801 acacaaccgc acatatttca gcaataagta caataaaaca ctcaaataaa aaaaacattt
     1861 taaatctctt tgtatttttg accgctgttt cgcgtaattt cacggtaaaa ctctggaaat
     1921 ctccactaca ttcctctcag cggctcctct caatgacagc tgaagaagtg acgcggctgc
     1981 ctgctgtgtt ttgattggtc gaattcactg gaggcttcca gaacagtgta gagtctgaac
     2041 gggtgcgcgc tctgctgtat ttaaagggcg aaagagagac cgcagagaaa ctcaaccgaa
     2101 gagaagcgac ttgacaaaga agaaaagagc agcctgacag gactttttcc ccgacgaggt
     2161 gtttattcgc tctatttaag aatctactgt aaggtaagtc tcaatatatt gtactctatt
     2221 ggctaatcag aattatatag agattatatg tacttaatgt caaaaaatca actttgtata
     2281 tgtaatcttt ttacatgtgg actgcctatg ttcatcttat tttaggtcta ctagaaaatt
     2341 atatttcccg ttttcacaat aaggattttt aaaaaaagca atgaacagac gggcatttac
     2401 tttatgttgc tgacattatt ttatatgagc ataataacca taaatactag caaatgtcct
     2461 aaatgaattt gtgttaatgt tgtctacaaa agaaaattag cgttttactt gtacaactaa
     2521 taataacttg gttattaaga gaatttcact tgttgactag aaaaatcctt tcataatgaa
     2581 acaattgcac cataaattgt ataaatataa aattaattct aattgttttt ttttcctgcc
     2641 aagtttgtac aaaaaagcag gcttgtaaaa cgacggccag tgagcgcgcg taatacgact
     2701 cactataggg cgaattgggt accgggcccc ccctcgaggt cgacgccacc atggcttctc
     2761 cacctaagaa gaagagaaag gtgggaagca tggataagaa gtatagcatc ggcctggata
     2821 ttggaactaa ctccgtgggt tgggcagtga ttacagacga ctacaaggtc cctagcaaga
     2881 aatttaaggt gctgggtaac accgacaggc acagcatcaa gaaaaatctg attggagccc
     2941 tgctgttcgg ttctggagag actgccgaag caacacgcct gaaaagaaca gcaagaaggc
     3001 gctataccag aaggaagaat agaatctgtt acctgcagga gattttctct aacgaaatgg
     3061 ctaaggtgga cgattcattc tttcataggc tggaggaaag tttcctggtc gaggaagata
     3121 agaaacacga gcgccatcct atctttggaa acattgtgga cgaggtcgcc tatcacgaaa
     3181 aatacccaac catctatcat ctgcgcaaga aactggctga ctctactgat aaagccgacc
     3241 tgagactgat ctatctggct ctggcccaca tgattaagtt caggggtcat tttctgatcg
     3301 agggcgatct gaaccccgac aattccgatg tggacaagct gttcatccag ctggtccaga
     3361 tttacaatca gctgtttgag gaaaacccta ttaatgcttc cagagtggac gcaaaagcta
     3421 tcctgtcagc caggctgtcc aagtcacgca gactggagaa cctgattgca cagctgcccg
     3481 gagaaaagag gaacggtctg tttggaaatc tgatcgctct gagtctgggc ctgactccta
     3541 acttcaaaag caattttgat ctggctgagg acgccaaact gcagctgtca aaggacacat
     3601 atgacgatga cctggataac ctgctggcac agatcggaga tcagtacgct gacctgttcc
     3661 tggctgccaa aaatctgtcc gacgcaatcc tgctgtcaga tattctgaga gtgaacagcg
     3721 agattacaaa agcacctctg agtgccagca tgatcaagag atatgacgag caccatcagg
     3781 atctgaccct gctgaaggct ctggtcaggc agcagctgcc agagaagtac aaggaaattt
     3841 tctttgatca gtccaagaac ggctacgccg gttatatcga cggaggcgca tcacaggagg
     3901 aattctacaa gtttatcaaa cctattctgg agaagatgga cggaactgag gaactgctgg
     3961 tgaaactgaa tagagaggac ctgctgagga agcagcgcac atttgataac ggttccatcc
     4021 cacaccagat tcatctggga gagctgcacg ctatcctgag gcgccaggaa gacttctacc
     4081 cctttctgaa agataaccgc gagaagatcg aaaaaattct gaccttcaga atcccttact
     4141 atgtgggtcc actggctcgc ggaaacagca gatttgcctg gatgactcgc aaatccgagg
     4201 aaaccattac tccttggaac ttcgaggaag tggtcgataa gggcgcctct gcacagtcct
     4261 tcatcgagag aatgactaat tttgacaaaa acctgcccaa tgagaaagtg ctgcctaagc
     4321 actccctgct gtacgagtat ttcactgtct ataacgaact gacaaaggtg aaatacgtca
     4381 ccgagggcat gagaaagcca gccttcctgt caggagagca gaagaaagca atcgtggatc
     4441 tgctgtttaa aaccaacagg aaagtgactg tcaagcagct gaaggaggac tacttcaaga
     4501 aaattgaatg cttcgattcc gtggagatca gcggagtcga agacagattt aacgcaagcc
     4561 tgggcgctta ccacgatctg ctgaagatca ttaaggataa agacttcctg gacaacgagg
     4621 aaaatgagga tatcctggaa gacattgtgc tgacactgac cctgtttgag gacagaggaa
     4681 tgatcgagga aagactgaaa acctatgctc atctgttcga tgacaaggtg atgaaacagc
     4741 tgaagagaag gcgctacact ggctggggta gactgagcag gaagctgatc aacggcatta
     4801 gggataaaca gtcaggaaag acaatcctgg actttctgaa aagtgatggc ttcgccaacc
     4861 gcaattttat gcagctgatt cacgatgaca gtctgacctt caaagaggac atccagaagg
     4921 ctcaggtgtc tggacagggc cactccctgc atgagcagat tgcaaacctg gctggaagcc
     4981 cagccatcaa gaaaggcatt ctgcagacag tgaaaatcgt cgatgagctg gtgaaagtca
     5041 tgggccataa gcccgaaaac atcgtgattg agatggctcg cgaaaatcag acaacccaga
     5101 agggtcagaa gaacagtaga gagaggatga aaagaatcga ggaaggcatt aaggagctgg
     5161 gtagccagat cctgaaagag cacccagtgg aaaacacaca gctgcagaat gagaagctgt
     5221 atctgtacta tctgcagaat ggaagagata tgtacgtgga ccaggagctg gatattaaca
     5281 ggctgtctga ttacgacgtg gatcatatcg tcccccagag tttcatcaaa gatgacagca
     5341 ttgacaacaa ggtgctgacc aggtccgaca aaaacagagg aaaatcagat aatgtcccta
     5401 gtgaggaagt ggtcaagaaa atgaagaact actggagaca gctgctgaat gccaaactga
     5461 tcactcagag gaagtttgat aacctgacaa aagcagagcg cggtggactg tcagaactgg
     5521 acaaagctgg attcatcaag aggcagctgg tggaaacacg ccagatcact aaacacgtcg
     5581 cacagattct ggatagtcgc atgaacacaa agtacgatga gaatgacaaa ctgatcagag
     5641 aagtgaaggt cattaccctg aagagtaaac tggtcagcga ctttaggaaa gatttccagt
     5701 tttataaggt ccgcgagatt aacaattatc accatgccca tgacgcatac ctgaacgccg
     5761 tggtcggtac cgcactgatc aagaaatacc caaaactgga gagcgaattc gtgtacggag
     5821 actataaggt gtacgatgtc agaaaaatga tcgccaagtc cgagcaggaa attggaaaag
     5881 ctactgccaa gtatttcttt tactcaaaca tcatgaattt ctttaagaca gagatcaccc
     5941 tggccaatgg agaaatccgc aaaaggcccc tgattgagac aaacggagag acaggcgaaa
     6001 tcgtgtggga caaaggcaga gattttgcaa ccgtgaggaa ggtcctgagc atgcctcaag
     6061 tgaatatcgt caagaaaact gaggtgcaga caggcggttt ctcaaaagaa agtattctgc
     6121 caaaacgcaa ctctgataag ctgatcgcta gaaagaaaga ctgggaccct aagaagtatg
     6181 gaggctttga ctctcccact gtggcatact ccgtcctggt ggtcgctaag gtggagaagg
     6241 gcaaaagcaa gaaactgaaa tctgtcaagg agctgctggg tatcacaatt atggagagaa
     6301 gctctttcga gaagaaccca atcgattttc tggaggccaa aggttataag gaagtgaaga
     6361 aagacctgat cattaaactg cccaagtaca gtctgtttga gctggaaaac ggcaggaaac
     6421 gcatgctggc aagcgctgga gagctgcaga aaggcaatga actggccctg ccttctaagt
     6481 acgtgaactt cctgtatctg gcaagccact acgagaagct gaaaggatct ccagaggata
     6541 acgaacagaa acagctgttt gtggagcagc acaagcatta tctggacgag atcattgaac
     6601 agattagcga gttctctaaa agagtgatcc tggccgacgc aaatctggat aaggtcctgt
     6661 ctgcttacaa caaacacaga gataagccca tcagggagca ggccgaaaat atcattcatc
     6721 tgttcactct gacaaacctg ggcgcacctg cagctttcaa gtacttcgac actacaatcg
     6781 atagaaagag gtacacctcc actaaggagg tgctggacgc tacactgatc catcagagta
     6841 ttaccggcct gtacgaaaca aggattgacc tgtctcagct gggtggcgac tcaccggtgg
     6901 gatcccctaa gaagaagaga aaggtgaggt ctggtggcgg agagggcaga ggcagtctgc
     6961 tgacatgcgg tgacgtggaa gagaatcccg gcccttctag aatggtgagc aagggcgaag
     7021 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     7081 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     7141 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     7201 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     7261 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     7321 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     7381 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     7441 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     7501 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     7561 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     7621 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     7681 ccgccgggat cactctcggc atggacgagc tgtacaagta agcggccgcc accgcggtgg
     7741 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     7801 tagcacccag ctttcttgta caaagtgggg gatccagaca tgataagata cattgatgag
     7861 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     7921 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     7981 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttccaa ctttattata
     8041 catagttgat aattcactgg ccgtcgtttt acggtaccgc acactattaa tggtacgaac
     8101 tttaaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt
     8161 aaacttgcta tgctgtttcc agcatagctc ttaaactagt cgactgtcgt gcagtcgaac
     8221 tgagagccgg aagaagagct atatatacca gggaattctg ggtatgtttc ttactggtgg
     8281 taagtagata caccccttat ttaattagct tccaacaaat ttgaatgtat acactggtga
     8341 gatagacctg taaactctta tgatctgcaa aattgcaagc caatgttttt gtggaaggtt
     8401 tattttgagt atttcctcag ctgtagacca catgtaaaaa aggttcctct ctgctagctg
     8461 tgattggatg tttcggtgtg acagtgaaga acccgtggcc acaatgttcc acaccctccc
     8521 ccacagacat gcgtagaatg tttccccctc aacgatgacg agaacaaaag gttgaaagac
     8581 cagcaaatgc tcctttcaca accaccaaat cacaggttta aaaaaagcac cgactcggtg
     8641 ccactttttc aagttgataa cggactagcc ttatttaaac ttgctatgct gtttccagca
     8701 tagctcttaa actgacagca gatactgctg tcgaactagg agcctggaga actgctatat
     8761 aaacccggga cttctgggta tgtttctgcc atgtggtaag tggataaacc tacttattca
     8821 actcatttac ctcaaacatg acagtaaacc tcttcggggc aaacctgcaa acgcttaaaa
     8881 ccttcaaagt ttgaaaccag cagttttgta gaggtttacc gtagtatttt tgaactgtaa
     8941 agcatatgca gaggaagctg gttttaggct tgtgtgattg gatgagttct tctgtgacgt
     9001 aactgatctt ttgtcctcgt gtccttcgcc ttcccccaca ggcatgcgcg gaccttcccc
     9061 cgccgcagac cagaggacaa catgctcaga cgccagcaaa cggcccatcg aaacattcac
     9121 agactctcgg attggtttaa aaaaagcacc gactcggtgc cactttttca agttgataac
     9181 ggactagcct tatttaaact tgctatgctg tttccagcat agctcttaaa ctactgcatg
     9241 atatctctgt cgaactggga gtctggagga cggctatata taccagtgac ttctgggtat
     9301 gtgtttcatg tgtggtatat ggcttactta cttattcaac tcatttacac ttacatgtgt
     9361 cagtaaatca ctgtgtagta gtaaacctgt aaacccttaa attctttaaa atttgaaaac
     9421 cagcagttta tgtaggttta ccgtagagtt ttggagctgt aagcgtatgc gaaggaaaca
     9481 ggccgtttct ttagagtgat tggatgagtt cctgaacttc ctgtgtgaca tcgaggagct
     9541 tttgttcacg tgtccttctc ctccccccac aggcatgcgc agatcccccc tgccttggat
     9601 aagagcacaa aacactccga caaaaggaag ctgctctcac tagagcagag cccatgaaaa
     9661 cccagacaga cacgtggagg gtaatgagtt taaaaaaagc accgactcgg tgccactttt
     9721 tcaagttgat aacggactag ccttatttaa acttgctatg ctgtttccag catagctctt
     9781 aaactagtcg tagcgactgt agtcgaacca agagctggag ggagagctat atataccagg
     9841 gacttctggg tatgtttttg ggaggtggtg agtgactaaa ccacttattc agctccctta
     9901 gatcaagtct gaccctatat catggtgaca taaacctgca aactgataaa acctgaagga
     9961 tctcaaatcc agagtttgtg tgagggatta ccgtggtaat tttcaaactg taaagcatat
    10021 gcaaaattat ctggtcttgg cttgagtgat tgggtgtctc ggtgtgatgc agggacgttt
    10081 tcagtgacgt gtccttctcc ctcccccaca ggcatgcgca gaacatttcc cccctccttg
    10141 aagaccagaa caaaagacgc cgagagcagg aaactcgtct tactgaatga ccgaggctgg
    10201 agaaacgctg agagacttga ctaaaaaagc accgactcgg tgccactttt tcaagttgat
    10261 aacggactag ccttatttaa acttgctatg ctgtttccag catagctctt aaactactcg
    10321 atgtctatgc tctcgaacca agagctggag ggagagctat atataccagg gacttctggg
    10381 tatgtttttg ggaggtggtg agtgactaaa ccacttattc agctccctta gatcaagtct
    10441 gaccctatat catggtgaca taaacctgca aactgataaa acctgaagga tctcaaatcc
    10501 agagtttgtg tgagggatta ccgtggtaat tttcaaactg taaagcatat gcaaaattat
    10561 ctggtcttgg cttgagtgat tgggtgtctc ggtgtgatgc agggacgttt tcagtgacgt
    10621 gtccttctcc ctcccccaca ggcatgcgca gaacatttcc cccctccttg aagaccagaa
    10681 caaaagacgc cgagagcagg aaactcgtct tactgaatga ccgaggctgg agaaacgctg
    10741 agagacttac tgatgcattt catttaatgt atcgatgatg atccagacat gataagatac
    10801 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
    10861 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
    10921 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
    10981 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca gatctgcgaa
    11041 gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg
    11101 ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata atttccctaa
    11161 tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag
    11221 gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat
    11281 aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg
    11341 gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta
    11401 aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga
    11461 tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc
    11521 aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc
    11581 cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
    11641 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    11701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    11761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    11821 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    11881 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    11941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    12001 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    12061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    12121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    12181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    12241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    12301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    12361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    12421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    12481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    12541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    12601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    12661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    12721 cacgttaagg cattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    12781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    12841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    12901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    12961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    13021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    13081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    13141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    13201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    13261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    13321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    13381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    13441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    13501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    13561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    13621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    13681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    13741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    13801 cgcgcacatt tccccgaaaa g
//
