LOCUS       sequence-211937-        9760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-GFP fusion protein under the regulation of chicken beta-actin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9760)
  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 9760)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..9760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..654
                     /label=chicken beta-actin promoter
     intron          655..1671
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1595..1617
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1679..1698
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1777..1797
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1798..5898
                     /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             1798..5898
                     /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             5908..5928
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5938..5940,5941..6654)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5938..5940,5941..6651)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5980..6001)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6241..6260)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6588..6609
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6748..6767)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6813..6868
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6867..6886)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7259..7275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7259..7275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7272..7294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7283..7299
                     /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(7307..7313,7314..7331,7332..7337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7352..7373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7432..7627
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7478..7613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7633..7767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7670..7689)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7724..7743
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7835..7852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8006..8594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8086..8105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8765..9556,9557..9625))
                     /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     9388..9407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9626..9730)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcgg gggggggggg
      541 ggggggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      601 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      661 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      781 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
      841 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
      901 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
      961 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1081 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1321 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1381 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1441 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1621 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattgctcga gagttcgaag
     1741 ccttaagtac gcgtaggcgc gccaaccatg gctagcccca aaaagaagag gaaagtggac
     1801 aagaagtatt ctatcggact ggacatcggg actaatagcg tcgggtgggc cgtgatcact
     1861 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc
     1921 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc
     1981 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt
     2041 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa
     2101 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc
     2161 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg
     2221 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc
     2281 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat
     2341 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat
     2401 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt
     2461 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt
     2521 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc
     2581 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc
     2641 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg
     2701 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt
     2761 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag
     2821 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac
     2881 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag
     2941 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag
     3001 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc
     3061 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa
     3121 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt
     3181 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg
     3241 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc
     3301 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac
     3361 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc
     3421 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag
     3481 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg
     3541 gtcgaagatc ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag
     3601 gacaaggatt ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact
     3661 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg
     3721 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt
     3781 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc
     3841 cttaagagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg
     3901 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag
     3961 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa
     4021 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag
     4081 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa
     4141 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa
     4201 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg
     4261 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga ccacatcgtc
     4321 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag
     4381 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat
     4441 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag
     4501 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg
     4561 gagactcggc agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag
     4621 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg
     4681 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat
     4741 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca
     4801 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt
     4861 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc
     4921 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc
     4981 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc
     5041 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc
     5101 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg
     5161 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct
     5221 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa
     5281 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc
     5341 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc
     5401 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag
     5461 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac
     5521 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat
     5581 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc
     5641 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt
     5701 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc
     5761 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg
     5821 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc
     5881 tcacagctcg gaggggatga gggagctccc aagaaaaagc gcaaggtagg tagttccgtg
     5941 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     6001 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     6061 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     6121 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     6181 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     6241 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     6301 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     6361 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     6421 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     6481 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     6541 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     6601 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgagcg
     6661 gccgcaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     6721 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     6781 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     6841 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     6901 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     6961 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     7021 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     7081 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     7141 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     7201 cctgcaccgt cgaccagctg gtcgacggtg caccgtcgac cagcttggcg taatcatggt
     7261 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7321 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     7441 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7501 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7561 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     7621 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7681 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7741 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     7801 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7861 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7921 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7981 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8041 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     8221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8341 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8401 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     8461 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8521 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8581 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8641 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8701 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8761 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8821 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8881 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8941 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9001 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9061 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     9121 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     9181 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     9241 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9301 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9361 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9421 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9481 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9541 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9601 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9661 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9721 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
//
