LOCUS       Exported                9671 bp ds-DNA     circular SYN 16-JUN-2022
DEFINITION  Tissue-specific knock-out in zebrafish, Cas9 and GFP driven by a UAS
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pUAS:Cas9T2AGFP;U6:sgRNA1;U6:sgRNA2
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 Jun 16, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9671
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     179..198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(417..521)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(599..1054)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(736..757)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     948..967
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1180..1202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1194..1211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1195..1211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1218..1237
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1218..1236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1895..1989
                     /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)"
     promoter        2008..2246
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     2193..2214
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     regulatory      2261..2270
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2279..2299
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2306..6406
                     /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             6319..6327
                     /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             6422..6442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6458..6511
                     /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             6512..7231
                     /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             6512..7228
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6557..6578)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6818..6837)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7165..7186
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(7239..7256)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7285..7304)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7339..7358
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8030..8048)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8031..8048)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8066..8082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8066..8082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8079..8101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8090..8106
                     /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(8114..8144)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8159..8180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8297..8314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8468..9056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8548..8567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9227..416)
                     /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"
ORIGIN
        1 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
       61 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
      121 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
      181 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
      241 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
      301 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
      361 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      421 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      481 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      541 ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
      601 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
      661 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
      721 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
      781 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
      841 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
      901 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
      961 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     1021 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
     1081 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     1141 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     1201 cgacggccag tgaattgtaa tacgactcac tatagggcga attgggccca gaggtgtaaa
     1261 gtacttgagt aattttactt gattactgta cttaagtatt atttttgggg atttttactt
     1321 tacttgagta caattaaaaa tcaatacttt tacttttact taattacatt tttttagaaa
     1381 aaaaagtact ttttactcct tacaatttta tttacagtca aaaagtactt attttttgga
     1441 gatcacttca ttctattttc ccttgctatt accaaaccaa ttgaattgcg ctgatgccca
     1501 gtttaattta aatagatccc atgggtcgac tacgcttaca gctccaaaac tctacggtaa
     1561 acctacataa actgctggtt ttcaaatttt aaagaattta agggtttaca ggtttactac
     1621 tacacagtga tttactgaca catgtaagtg taaatgagtt gaataagtaa gtaagccata
     1681 taccacacat gaaacacata cccagaagtc actggtatat atagccgtcc tccagactcc
     1741 cagctcgaga gaccgagaga gggtctcagt ttaagagcta tgctggaaac agcatagcaa
     1801 gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     1861 tggtcgacgg atccaagctt gcatgcctgc aggtcggagt actgtcctcc gagcggagta
     1921 ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg
     1981 tcctccgagc ggagactcta gcgagcgccg gagtataaat agaggcgctt cgtctacgga
     2041 gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc taagcaaata
     2101 aacaagcgca gctgaacaag ctaaacaatc tgcagtaaag tgcaagttaa agtgaatcaa
     2161 ttaaaagtaa ccagcaacca agtaaatcaa ctgcaactac tgaaatctgc caagaagtaa
     2221 ttattgaata caagaagaga actctgaata gggaattggg gccaccatgg cttctccacc
     2281 taagaagaag agaaaggtgg gaagcatgga taagaagtat agcatcggcc tggatattgg
     2341 aactaactcc gtgggttggg cagtgattac agacgactac aaggtcccta gcaagaaatt
     2401 taaggtgctg ggtaacaccg acaggcacag catcaagaaa aatctgattg gagccctgct
     2461 gttcggttct ggagagactg ccgaagcaac acgcctgaaa agaacagcaa gaaggcgcta
     2521 taccagaagg aagaatagaa tctgttacct gcaggagatt ttctctaacg aaatggctaa
     2581 ggtggacgat tcattctttc ataggctgga ggaaagtttc ctggtcgagg aagataagaa
     2641 acacgagcgc catcctatct ttggaaacat tgtggacgag gtcgcctatc acgaaaaata
     2701 cccaaccatc tatcatctgc gcaagaaact ggctgactct actgataaag ccgacctgag
     2761 actgatctat ctggctctgg cccacatgat taagttcagg ggtcattttc tgatcgaggg
     2821 cgatctgaac cccgacaatt ccgatgtgga caagctgttc atccagctgg tccagattta
     2881 caatcagctg tttgaggaaa accctattaa tgcttccaga gtggacgcaa aagctatcct
     2941 gtcagccagg ctgtccaagt cacgcagact ggagaacctg attgcacagc tgcccggaga
     3001 aaagaggaac ggtctgtttg gaaatctgat cgctctgagt ctgggcctga ctcctaactt
     3061 caaaagcaat tttgatctgg ctgaggacgc caaactgcag ctgtcaaagg acacatatga
     3121 cgatgacctg gataacctgc tggcacagat cggagatcag tacgctgacc tgttcctggc
     3181 tgccaaaaat ctgtccgacg caatcctgct gtcagatatt ctgagagtga acagcgagat
     3241 tacaaaagct cctctgagtg ccagcatgat caagagatat gacgagcacc atcaggatct
     3301 gaccctgctg aaggctctgg tcaggcagca gctgccagag aagtacaagg aaattttctt
     3361 tgatcagtcc aagaacggct acgccggtta tatcgacgga ggcgcatcac aggaggaatt
     3421 ctacaagttt atcaaaccta ttctggagaa gatggacgga actgaggaac tgctggtgaa
     3481 actgaataga gaggacctgc tgaggaagca gcgcacattt gataacggtt ccatcccaca
     3541 ccagattcat ctgggagagc tgcacgctat cctgaggcgc caggaagact tctacccctt
     3601 tctgaaagat aaccgcgaga agatcgaaaa aattctgacc ttcagaatcc cttactatgt
     3661 gggtccactg gctcgcggaa acagcagatt tgcctggatg actcgcaaat ccgaggaaac
     3721 cattactcct tggaacttcg aggaagtggt cgataagggc gcctctgcac agtccttcat
     3781 cgagagaatg actaattttg acaaaaacct gcccaatgag aaagtgctgc ctaagcactc
     3841 cctgctgtac gagtatttca ctgtctataa cgaactgaca aaggtgaaat acgtcaccga
     3901 gggcatgaga aagccagcct tcctgtcagg agagcagaag aaagcaatcg tggatctgct
     3961 gtttaaaacc aacaggaaag tgactgtcaa gcagctgaag gaggactact tcaagaaaat
     4021 tgaatgcttc gattccgtgg agatcagcgg agtcgaagac agatttaacg caagcctggg
     4081 cgcttaccac gatctgctga agatcattaa ggataaagac ttcctggaca acgaggaaaa
     4141 tgaggatatc ctggaagaca ttgtgctgac actgaccctg tttgaggaca gaggaatgat
     4201 cgaggaaaga ctgaaaacct atgctcatct gttcgatgac aaggtgatga aacagctgaa
     4261 gagaaggcgc tacactggct ggggtagact gagcaggaag ctgatcaacg gcattaggga
     4321 taaacagtca ggaaagacaa tcctggactt tctgaaaagt gatggcttcg ccaaccgcaa
     4381 ttttatgcag ctgattcacg atgacagtct gaccttcaaa gaggacatcc agaaggctca
     4441 ggtgtctgga cagggccact ccctgcatga gcagattgca aacctggctg gaagcccagc
     4501 catcaagaaa ggcattctgc agacagtgaa aatcgtcgat gagctggtga aagtcatggg
     4561 ccataagccc gaaaacatcg tgattgagat ggctcgcgaa aatcagacaa cccagaaggg
     4621 tcagaagaac agtagagaga ggatgaaaag aatcgaggaa ggcattaagg agctgggtag
     4681 ccagatcctg aaagagcacc cagtggaaaa cacacagctg cagaatgaga agctgtatct
     4741 gtactatctg cagaatggaa gagatatgta cgtggaccag gagctggata ttaacaggct
     4801 gtctgattac gacgtggatc atatcgtccc ccagagtttc atcaaagatg acagcattga
     4861 caacaaggtg ctgaccaggt ccgacaaaaa cagaggaaaa tcagataatg tccctagtga
     4921 ggaagtggtc aagaaaatga agaactactg gagacagctg ctgaatgcca aactgatcac
     4981 tcagaggaag tttgataacc tgacaaaagc agagcgcggt ggactgtcag aactggacaa
     5041 agctggattc atcaagaggc agctggtgga aacacgccag atcactaaac acgtcgcaca
     5101 gattctggat agtcgcatga acacaaagta cgatgagaat gacaaactga tcagagaagt
     5161 gaaggtcatt accctgaaga gtaaactggt cagcgacttt aggaaagatt tccagtttta
     5221 taaggtccgc gagattaaca attatcacca tgcccatgac gcatacctga acgccgtggt
     5281 cggtaccgca ctgatcaaga aatacccaaa actggagagc gaattcgtgt acggagacta
     5341 taaggtgtac gatgtcagaa aaatgatcgc caagtccgag caggaaattg gaaaagctac
     5401 tgccaagtat ttcttttact caaacatcat gaatttcttt aagacagaga tcaccctggc
     5461 caatggagaa atccgcaaaa ggcccctgat tgagacaaac ggagagacag gcgaaatcgt
     5521 gtgggacaaa ggcagagatt ttgcaaccgt gaggaaggtc ctgagcatgc ctcaagtgaa
     5581 tatcgtcaag aaaactgagg tgcagacagg cggtttctca aaagaaagta ttctgccaaa
     5641 acgcaactct gataagctga tcgctagaaa gaaagactgg gaccctaaga agtatggagg
     5701 ctttgactct cccactgtgg catactccgt cctggtggtc gctaaggtgg agaagggcaa
     5761 aagcaagaaa ctgaaatctg tcaaggagct gctgggtatc acaattatgg agagaagctc
     5821 tttcgagaag aacccaatcg attttctgga ggccaaaggt tataaggaag tgaagaaaga
     5881 cctgatcatt aaactgccca agtacagtct gtttgagctg gaaaacggca ggaaacgcat
     5941 gctggcaagc gctggagagc tgcagaaagg caatgaactg gccctgcctt ctaagtacgt
     6001 gaacttcctg tatctggcaa gccactacga gaagctgaaa ggatctccag aggataacga
     6061 acagaaacag ctgtttgtgg agcagcacaa gcattatctg gacgagatca ttgaacagat
     6121 tagcgagttc tctaaaagag tgatcctggc cgacgcaaat ctggataagg tcctgtctgc
     6181 ttacaacaaa cacagagata agcccatcag ggagcaggcc gaaaatatca ttcatctgtt
     6241 cactctgaca aacctgggcg cacctgcagc tttcaagtac ttcgacacta caatcgatag
     6301 aaagaggtac acctccacta aggaggtgct ggacgctaca ctgatccatc agagtattac
     6361 cggcctgtac gaaacaagga ttgacctgtc tcagctgggt ggcgactcac cggtgagatc
     6421 tcctaagaag aagagaaagg tgaggtccgg cggcggagag ggcagaggaa gtcttctaac
     6481 atgcggtgac gtggaggaga atcccggccc aatggtgagc aagggcgagg agctgttcac
     6541 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6601 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6661 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6721 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     6781 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     6841 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6901 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     6961 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7021 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     7081 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7141 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     7201 cactctcggc atggacgagc tgtacaagta atctagaact atagtgagtc gtattacgta
     7261 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     7321 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     7381 tgcaataaac cttgtacaaa gtgggggatc ctacgcttac agctccaaaa ctctacggta
     7441 aacctacata aactgctggt tttcaaattt taaagaattt aagggtttac aggtttacta
     7501 ctacacagtg atttactgac acatgtaagt gtaaatgagt tgaataagta agtaagccat
     7561 ataccacaca tgaaacacat acccagaagt cactggtata tatagccgtc ctccagactc
     7621 ccagctcgag acgattagat ctaatgcgtc tcgtttaaga gctatgctgg aaacagcata
     7681 gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     7741 tttttcaatg atgaattaat tcgagctcca ccgcggaagc ttaaacaaga atctctagtt
     7801 ttctttcttg cttttacttt tacttcctta atactcaagt acaattttaa tggagtactt
     7861 ttttactttt actcaagtaa gattctagcc agatactttt acttttaatt gagtaaaatt
     7921 ttccctaagt acttgtactt tcacttgagt aaaatttttg agtacttttt acacctctgc
     7981 tcgaccatat gggagagctc ccaacgcgtt ggatgcatag cttgagtatt ctatagtgtc
     8041 acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     8101 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     8161 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     8221 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     8281 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     8341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     8401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     8701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     9001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     9061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggca
     9121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     9181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     9241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     9301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9361 taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9661 gtcctccgat c
//
