LOCUS       sequence-472875-       10016 bp    DNA     circular SYN 17-NOV-2025
DEFINITION  Expression of nuclear GFP and nuclear Cas9 specifically in 
            cardiomyocytes driven by the cmlc2(myl7) promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10016)
  AUTHORS   Keeley S, Fernandez-Lajarin M, Bergemann D, John N, Parrott L, 
            Andrea BE, Gonzalez-Rosa JM
  TITLE     Rapid and robust generation of cardiomyocyte-specific crispants in 
            zebrafish using the cardiodeleter system.
  JOURNAL   Cell Rep Methods. 2025 Mar 24;5(3):101003. doi: 
            10.1016/j.crmeth.2025.101003.
  PUBMED    40132543
REFERENCE   2  (bases 1 to 10016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep 
            Methods."; date: "2025-03-24"; volume: "5(3):101003. doi"; pages: " 
            10.1016/j.crmeth.2025.101003"
COMMENT     Sequence Label: pTol2-cmlc2-nGFP-nCas9
FEATURES             Location/Qualifiers
     source          1..10016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     regulatory      1895..1904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1907..1927
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1973..1975,1976..1978,1979..2689)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1973..1975,1976..1978,1979..2689)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2018..2039)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2279..2298)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2626..2647
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2699..2755
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2768..2788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2795..6895
                     /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             6808..6816
                     /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             6911..6931
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6973..7107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6997..7016)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7051..7070
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7779..7795)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7779..7795)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7832..7852)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7832..7850)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7871..7887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7871..7887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7884..7906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7895..7911
                     /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(7919..7925,7926..7943,7944..7949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7964..7985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8102..8119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8273..8861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8353..8372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9032..9823,9824..9892))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGTGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9655..9674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9893..9997)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaatc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatc tccaaatcag cagacttaac attctttaaa
     1081 atgattgatt caatagtgat aaaaatcagg catagccagt tgtaacttta gataaattac
     1141 agaaaatgtc aaatacagag aaccgattct tttttatgat acatccaagc acacatttaa
     1201 cacaatccag gcaaaccccg aatttcacag tcacaagcac tgtttgtaca agagctttgc
     1261 ctaaggacac acagtctcta taagtccagg tcgttggttt cactcttatt ttaaacatgt
     1321 gacatttttc ctgccatcct gtcttaggct gctgtttgct tcattccatg tcacattaat
     1381 ttcctcagta gcacctttta cacacacagc caatcttttc cagaaaattc aattgctttg
     1441 aaaagataat gtgtgaacaa atccatttag aaaaggaaaa ttaagaattt gtaaaatcat
     1501 ctgtaaattg ttggcattct tctgtatatg aacatcacat catttacagg taaaggtctg
     1561 gtcattaatt atatgacaat ttactggtat tattttgtga aaggggctat tttcaatgca
     1621 ttcatccatc cttttcatcc cccaaatctc tcattcatgt ccccctcccc atctgcacac
     1681 tttatctcat tttccaccct gctggaatct gagcacttgt gcagttatca gggctcctgt
     1741 atttaggagg ctctgggtgt ccatgtaggg gacgaacaga aacactgcag acctttatag
     1801 aagaacaaat gataagagtc ctcatacata aagactccat tagaaacgtc agtgacccag
     1861 gagcccagac caacagcaaa gcagacagtg aaccgccacc atggctccaa agaagaagcg
     1921 taaggtaaat cgatcgaatt caaggcctct cgagcctcta gaaggtggcg gaatggtgag
     1981 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2041 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2101 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2161 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2221 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2281 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2341 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2401 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2461 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2521 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2581 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2641 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gaagcggagc
     2701 tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg gacctatggc
     2761 ttctccacct aagaagaaga gaaaggtggg aagcatggat aagaagtata gcatcggcct
     2821 ggatattgga actaactccg tgggttgggc agtgattaca gacgactaca aggtccctag
     2881 caagaaattt aaggtgctgg gtaacaccga caggcacagc atcaagaaaa atctgattgg
     2941 agccctgctg ttcggttctg gagagactgc cgaagcaaca cgcctgaaaa gaacagcaag
     3001 aaggcgctat accagaagga agaatagaat ctgttacctg caggagattt tctctaacga
     3061 aatggctaag gtggacgatt cattctttca taggctggag gaaagtttcc tggtcgagga
     3121 agataagaaa cacgagcgcc atcctatctt tggaaacatt gtggacgagg tcgcctatca
     3181 cgaaaaatac ccaaccatct atcatctgcg caagaaactg gctgactcta ctgataaagc
     3241 cgacctgaga ctgatctatc tggctctggc ccacatgatt aagttcaggg gtcattttct
     3301 gatcgagggc gatctgaacc ccgacaattc cgatgtggac aagctgttca tccagctggt
     3361 ccagatttac aatcagctgt ttgaggaaaa ccctattaat gcttccagag tggacgcaaa
     3421 agctatcctg tcagccaggc tgtccaagtc acgcagactg gagaacctga ttgcacagct
     3481 gcccggagaa aagaggaacg gtctgtttgg aaatctgatc gctctgagtc tgggcctgac
     3541 tcctaacttc aaaagcaatt ttgatctggc tgaggacgcc aaactgcagc tgtcaaagga
     3601 cacatatgac gatgacctgg ataacctgct ggcacagatc ggagatcagt acgctgacct
     3661 gttcctggct gccaaaaatc tgtccgacgc aatcctgctg tcagatattc tgagagtgaa
     3721 cagcgagatt acaaaagctc ctctgagtgc cagcatgatc aagagatatg acgagcacca
     3781 tcaggatctg accctgctga aggctctggt caggcagcag ctgccagaga agtacaagga
     3841 aattttcttt gatcagtcca agaacggcta cgccggttat atcgacggag gcgcatcaca
     3901 ggaggaattc tacaagttta tcaaacctat tctggagaag atggacggaa ctgaggaact
     3961 gctggtgaaa ctgaatagag aggacctgct gaggaagcag cgcacatttg ataacggttc
     4021 catcccacac cagattcatc tgggagagct gcacgctatc ctgaggcgcc aggaagactt
     4081 ctaccccttt ctgaaagata accgcgagaa gatcgaaaaa attctgacct tcagaatccc
     4141 ttactatgtg ggtccactgg ctcgcggaaa cagcagattt gcctggatga ctcgcaaatc
     4201 cgaggaaacc attactcctt ggaacttcga ggaagtggtc gataagggcg cctctgcaca
     4261 gtccttcatc gagagaatga ctaattttga caaaaacctg cccaatgaga aagtgctgcc
     4321 taagcactcc ctgctgtacg agtatttcac tgtctataac gaactgacaa aggtgaaata
     4381 cgtcaccgag ggcatgagaa agccagcctt cctgtcagga gagcagaaga aagcaatcgt
     4441 ggatctgctg tttaaaacca acaggaaagt gactgtcaag cagctgaagg aggactactt
     4501 caagaaaatt gaatgcttcg attccgtgga gatcagcgga gtcgaagaca gatttaacgc
     4561 aagcctgggc gcttaccacg atctgctgaa gatcattaag gataaagact tcctggacaa
     4621 cgaggaaaat gaggatatcc tggaagacat tgtgctgaca ctgaccctgt ttgaggacag
     4681 aggaatgatc gaggaaagac tgaaaaccta tgctcatctg ttcgatgaca aggtgatgaa
     4741 acagctgaag agaaggcgct acactggctg gggtagactg agcaggaagc tgatcaacgg
     4801 cattagggat aaacagtcag gaaagacaat cctggacttt ctgaaaagtg atggcttcgc
     4861 caaccgcaat tttatgcagc tgattcacga tgacagtctg accttcaaag aggacatcca
     4921 gaaggctcag gtgtctggac agggccactc cctgcatgag cagattgcaa acctggctgg
     4981 aagcccagcc atcaagaaag gcattctgca gacagtgaaa atcgtcgatg agctggtgaa
     5041 agtcatgggc cataagcccg aaaacatcgt gattgagatg gctcgcgaaa atcagacaac
     5101 ccagaagggt cagaagaaca gtagagagag gatgaaaaga atcgaggaag gcattaagga
     5161 gctgggtagc cagatcctga aagagcaccc agtggaaaac acacagctgc agaatgagaa
     5221 gctgtatctg tactatctgc agaatggaag agatatgtac gtggaccagg agctggatat
     5281 taacaggctg tctgattacg acgtggatca tatcgtcccc cagagtttca tcaaagatga
     5341 cagcattgac aacaaggtgc tgaccaggtc cgacaaaaac agaggaaaat cagataatgt
     5401 ccctagtgag gaagtggtca agaaaatgaa gaactactgg agacagctgc tgaatgccaa
     5461 actgatcact cagaggaagt ttgataacct gacaaaagca gagcgcggtg gactgtcaga
     5521 actggacaaa gctggattca tcaagaggca gctggtggaa acacgccaga tcactaaaca
     5581 cgtcgcacag attctggata gtcgcatgaa cacaaagtac gatgagaatg acaaactgat
     5641 cagagaagtg aaggtcatta ccctgaagag taaactggtc agcgacttta ggaaagattt
     5701 ccagttttat aaggtccgcg agattaacaa ttatcaccat gcccatgacg catacctgaa
     5761 cgccgtggtc ggtaccgcac tgatcaagaa atacccaaaa ctggagagcg aattcgtgta
     5821 cggagactat aaggtgtacg atgtcagaaa aatgatcgcc aagtccgagc aggaaattgg
     5881 aaaagctact gccaagtatt tcttttactc aaacatcatg aatttcttta agacagagat
     5941 caccctggcc aatggagaaa tccgcaaaag gcccctgatt gagacaaacg gagagacagg
     6001 cgaaatcgtg tgggacaaag gcagagattt tgcaaccgtg aggaaggtcc tgagcatgcc
     6061 tcaagtgaat atcgtcaaga aaactgaggt gcagacaggc ggtttctcaa aagaaagtat
     6121 tctgccaaaa cgcaactctg ataagctgat cgctagaaag aaagactggg accctaagaa
     6181 gtatggaggc tttgactctc ccactgtggc atactccgtc ctggtggtcg ctaaggtgga
     6241 gaagggcaaa agcaagaaac tgaaatctgt caaggagctg ctgggtatca caattatgga
     6301 gagaagctct ttcgagaaga acccaatcga ttttctggag gccaaaggtt ataaggaagt
     6361 gaagaaagac ctgatcatta aactgcccaa gtacagtctg tttgagctgg aaaacggcag
     6421 gaaacgcatg ctggcaagcg ctggagagct gcagaaaggc aatgaactgg ccctgccttc
     6481 taagtacgtg aacttcctgt atctggcaag ccactacgag aagctgaaag gatctccaga
     6541 ggataacgaa cagaaacagc tgtttgtgga gcagcacaag cattatctgg acgagatcat
     6601 tgaacagatt agcgagttct ctaaaagagt gatcctggcc gacgcaaatc tggataaggt
     6661 cctgtctgct tacaacaaac acagagataa gcccatcagg gagcaggccg aaaatatcat
     6721 tcatctgttc actctgacaa acctgggcgc acctgcagct ttcaagtact tcgacactac
     6781 aatcgataga aagaggtaca cctccactaa ggaggtgctg gacgctacac tgatccatca
     6841 gagtattacc ggcctgtacg aaacaaggat tgacctgtct cagctgggtg gcgactcacc
     6901 ggtgagatct cctaagaaga agagaaaggt gtgaccgcgg aatctagact atagtgagtc
     6961 gtattataat aggatccaga catgataaga tacattgatg agtttggaca aaccacaact
     7021 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     7081 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     7141 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     7201 gctgattatg atcctctaga tcagatctgc gaagatacgg ccacgggtgc tcttgatcct
     7261 gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact tcctccattt
     7321 tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc tgtgcattgt
     7381 ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct gaccaatttc
     7441 atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta aactgggcat
     7501 cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa
     7561 aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa
     7621 aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa
     7681 aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag tactttacac
     7741 ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt ctagagcggc
     7801 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     7861 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7921 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7981 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     8041 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8101 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8161 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8221 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8281 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8341 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8401 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8461 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8521 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8581 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8641 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8701 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8761 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8821 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8881 ctacggggtc tgacgctcag tggaacgaaa actcacgtta aggcattttg gtcatgagat
     8941 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9001 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9061 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9121 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9181 gctcaccggt tccagattta tcagcaataa accagccagc cggaagtgcc gagcgcagaa
     9241 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9301 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9361 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9421 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9481 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9541 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9601 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9661 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9721 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9781 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9841 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9901 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9961 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtg
//
