LOCUS       sequence-211939-        9040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 via the chicken beta actin promoter for 
            electroporation in chicken embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9040)
  AUTHORS   Gandhi S, Piacentino ML, Vieceli FM, Bronner ME
  TITLE     Optimization of CRISPR/Cas9 genome editing for loss-of-function in 
            the early chick embryo.
  JOURNAL   Dev Biol. 2017 Dec 1;432(1):86-97. doi: 10.1016/j.ydbio.2017.08.036.
  PUBMED    29150011
REFERENCE   2  (bases 1 to 9040)
  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"; doi: 
            "10.1016/j.ydbio.2017.08"; journalName: "Dev Biol"; date: 
            "2017-12-1- 1"; volume: "432"; issue: "1"; pages: "86-97"
COMMENT     Sequence Label: pCAGG>nls-hCas9-nls
FEATURES             Location/Qualifiers
     source          1..9040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1784..1804
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1805..5905
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1805..5905
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5915..5935
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6032..6051)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6097..6152
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6151..6170)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6543..6559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6543..6559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6556..6578)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6567..6583
                     /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(6591..6597,6598..6615,6616..6621))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6636..6657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6716..6911
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6762..6897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6917..7051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6954..6973)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7008..7027
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7119..7136)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7290..7878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7370..7389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8049..8840,8841..8909))
                     /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     8672..8691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8910..9014)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgggggggg
      541 gggggggggg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgagag
     1741 ttcgaagcct taagtacgcg taggcgcgcc aaccatggct agccccaaaa agaagaggaa
     1801 agtggacaag aagtattcta tcggactgga catcgggact aatagcgtcg ggtgggccgt
     1861 gatcactgac gagtacaagg tgccctctaa gaagttcaag gtgctcggga acaccgaccg
     1921 gcattccatc aagaaaaatc tgatcggagc tctcctcttt gattcagggg agaccgctga
     1981 agcaacccgc ctcaagcgga ctgctagacg gcggtacacc aggaggaaga accggatttg
     2041 ttaccttcaa gagatattct ccaacgaaat ggcaaaggtc gacgacagct tcttccatag
     2101 gctggaagaa tcattcctcg tggaagagga taagaagcat gaacggcatc ccatcttcgg
     2161 taatatcgtc gacgaggtgg cctatcacga gaaataccca accatctacc atcttcgcaa
     2221 aaagctggtg gactcaaccg acaaggcaga cctccggctt atctacctgg ccctggccca
     2281 catgatcaag ttcagaggcc acttcctgat cgagggcgac ctcaatcctg acaatagcga
     2341 tgtggataaa ctgttcatcc agctggtgca gacttacaac cagctctttg aagagaaccc
     2401 catcaatgca agcggagtcg atgccaaggc cattctgtca gcccggctgt caaagagccg
     2461 cagacttgag aatcttatcg ctcagctgcc gggtgaaaag aaaaatggac tgttcgggaa
     2521 cctgattgct ctttcacttg ggctgactcc caatttcaag tctaatttcg acctggcaga
     2581 ggatgccaag ctgcaactgt ccaaggacac ctatgatgac gatctcgaca acctcctggc
     2641 ccagatcggt gaccaatacg ccgacctttt ccttgctgct aagaatcttt ctgacgccat
     2701 cctgctgtct gacattctcc gcgtgaacac tgaaatcacc aaggcccctc tttcagcttc
     2761 aatgattaag cggtatgatg agcaccacca ggacctgacc ctgcttaagg cactcgtccg
     2821 gcagcagctt ccggagaagt acaaggaaat cttctttgac cagtcaaaga atggatacgc
     2881 cggctacatc gacggaggtg cctcccaaga ggaattttat aagtttatca aacctatcct
     2941 tgagaagatg gacggcaccg aagagctcct cgtgaaactg aatcgggagg atctgctgcg
     3001 gaagcagcgc actttcgaca atgggagcat tccccaccag atccatcttg gggagcttca
     3061 cgccatcctt cggcgccaag aggacttcta cccctttctt aaggacaaca gggagaagat
     3121 tgagaaaatt ctcactttcc gcatccccta ctacgtggga cccctcgcca gaggaaatag
     3181 ccggtttgct tggatgacca gaaagtcaga agaaactatc actccctgga acttcgaaga
     3241 ggtggtggac aagggagcca gcgctcagtc attcatcgaa cggatgacta acttcgataa
     3301 gaacctcccc aatgagaagg tcctgccgaa acattccctg ctctacgagt actttaccgt
     3361 gtacaacgag ctgaccaagg tgaaatatgt caccgaaggg atgaggaagc ccgcattcct
     3421 gtcaggcgaa caaaagaagg caattgtgga ccttctgttc aagaccaata gaaaggtgac
     3481 cgtgaagcag ctgaaggagg actatttcaa gaaaattgaa tgcttcgact ctgtggagat
     3541 tagcggggtc gaagatcggt tcaacgcaag cctgggtacc taccatgatc tgcttaagat
     3601 catcaaggac aaggattttc tggacaatga ggagaacgag gacatccttg aggacattgt
     3661 cctgactctc actctgttcg aggaccggga aatgatcgag gagaggctta agacctacgc
     3721 ccatctgttc gacgataaag tgatgaagca acttaaacgg agaagatata ccggatgggg
     3781 acgccttagc cgcaaactca tcaacggaat ccgggacaaa cagagcggaa agaccattct
     3841 tgatttcctt aagagcgacg gattcgctaa tcgcaacttc atgcaactta tccatgatga
     3901 ttccctgacc tttaaggagg acatccagaa ggcccaagtg tctggacaag gtgactcact
     3961 gcacgagcat atcgcaaatc tggctggttc acccgctatt aagaagggta ttctccagac
     4021 cgtgaaagtc gtggacgagc tggtcaaggt gatgggtcgc cataaaccag agaacattgt
     4081 catcgagatg gccagggaaa accagactac ccagaaggga cagaagaaca gcagggagcg
     4141 gatgaaaaga attgaggaag ggattaagga gctcgggtca cagatcctta aagagcaccc
     4201 ggtggaaaac acccagcttc agaatgagaa gctctatctg tactaccttc aaaatggacg
     4261 cgatatgtat gtggaccaag agcttgatat caacaggctc tcagactacg acgtggacca
     4321 catcgtccct cagagcttcc tcaaagacga ctcaattgac aataaggtgc tgactcgctc
     4381 agacaagaac cggggaaagt cagataacgt gccctcagag gaagtcgtga aaaagatgaa
     4441 gaactattgg cgccagcttc tgaacgcaaa gctgatcact cagcggaagt tcgacaatct
     4501 cactaaggct gagaggggcg gactgagcga actggacaaa gcaggattca ttaaacggca
     4561 acttgtggag actcggcaga ttactaaaca tgtcgcccaa atccttgact cacgcatgaa
     4621 taccaagtac gacgaaaacg acaaacttat ccgcgaggtg aaggtgatta ccctgaagtc
     4681 caagctggtc agcgatttca gaaaggactt tcaattctac aaagtgcggg agatcaataa
     4741 ctatcatcat gctcatgacg catatctgaa tgccgtggtg ggaaccgccc tgatcaagaa
     4801 gtacccaaag ctggaaagcg agttcgtgta cggagactac aaggtctacg acgtgcgcaa
     4861 gatgattgcc aaatctgagc aggagatcgg aaaggccacc gcaaagtact tcttctacag
     4921 caacatcatg aatttcttca agaccgaaat cacccttgca aacggtgaga tccggaagag
     4981 gccgctcatc gagactaatg gggagactgg cgaaatcgtg tgggacaagg gcagagattt
     5041 cgctaccgtg cgcaaagtgc tttctatgcc tcaagtgaac atcgtgaaga aaaccgaggt
     5101 gcaaaccgga ggcttttcta aggaatcaat cctccccaag cgcaactccg acaagctcat
     5161 tgcaaggaag aaggattggg accctaagaa gtacggcgga ttcgattcac caactgtggc
     5221 ttattctgtc ctggtcgtgg ctaaggtgga aaaaggaaag tctaagaagc tcaagagcgt
     5281 gaaggaactg ctgggtatca ccattatgga gcgcagctcc ttcgagaaga acccaattga
     5341 ctttctcgaa gccaaaggtt acaaggaagt caagaaggac cttatcatca agctcccaaa
     5401 gtatagcctg ttcgaactgg agaatgggcg gaagcggatg ctcgcctccg ctggcgaact
     5461 tcagaagggt aatgagctgg ctctcccctc caagtacgtg aatttcctct accttgcaag
     5521 ccattacgag aagctgaagg ggagccccga ggacaacgag caaaagcaac tgtttgtgga
     5581 gcagcataag cattatctgg acgagatcat tgagcagatt tccgagtttt ctaaacgcgt
     5641 cattctcgct gatgccaacc tcgataaagt ccttagcgca tacaataagc acagagacaa
     5701 accaattcgg gagcaggctg agaatatcat ccacctgttc accctcacca atcttggtgc
     5761 ccctgccgca ttcaagtact tcgacaccac catcgaccgg aaacgctata cctccaccaa
     5821 agaagtgctg gacgccaccc tcatccacca gagcatcacc ggactttacg aaactcggat
     5881 tgacctctca cagctcggag gggatgaggg agctcccaag aaaaagcgca aggtataatg
     5941 agcggccgca attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     6001 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     6061 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     6121 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     6181 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     6241 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     6301 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     6361 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     6421 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     6481 tcgacctgca ccgtcgacca gctggtcgac ggtgcaccgt cgaccagctt ggcgtaatca
     6541 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6601 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6661 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     6721 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6781 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6841 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6901 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6961 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7021 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     7081 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8161 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8281 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8341 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8401 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8461 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8521 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8581 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8641 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8701 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8761 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8821 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8881 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9001 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
