LOCUS       sequence-307166-       10010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CAS9-eGFP fusion protein in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10010)
  AUTHORS   Fujii W, Kawasaki K, Sugiura K, Naito K
  TITLE     Efficient generation of large-scale genome-modified mice using gRNA 
            and CAS9 endonuclease.
  JOURNAL   Nucleic Acids Res. 2013 Aug 30.
  PUBMED    23997119
REFERENCE   2  (bases 1 to 10010)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2013 Aug 30."
COMMENT     Sequence Label: pCAG-T3-hCASeGFP-pA
FEATURES             Location/Qualifiers
     source          1..10010
                     /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"
     promoter        1764..1782
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1843..1866
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1876..1896
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1921..6021
                     /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             1921..6021
                     /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             1921..6021
                     /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             join(6028..6030,6031..6741)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6028..6030,6031..6741)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6070..6091)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6331..6350)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6678..6699
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6757..6777
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7032..7051)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7097..7152
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7151..7170)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7513..7529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7513..7529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7526..7548)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7537..7553
                     /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(7561..7567,7568..7585,7586..7591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7606..7627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7686..7881
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7732..7867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7887..8021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7924..7943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7978..7997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8089..8106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8260..8848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8340..8359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9019..9810,9811..9879))
                     /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     9642..9661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9880..9984)
                     /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 gggggggcgc gccggggggg gggggggggg gcggggggcg
      541 gggcggggcg 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 ggggctcggg gggggggcgc 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 tcgggcaacg tgctggttat tgtgctgtct
     1741 catcattttg gcaaagaatt cgaaattaac cctcactaaa gggaacaaaa gctggagctc
     1801 caccgcggtg gcggccgctc tagcccgggc ggatccacca tggattacaa ggatgacgac
     1861 gataagatca tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
     1921 gacaagaagt actccattgg gctcgatatc ggcacaaaca gcgtcggctg ggccgtcatt
     1981 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
     2041 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
     2101 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
     2161 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     2221 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     2281 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     2341 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     2401 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     2461 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     2521 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     2581 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     2641 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     2701 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     2761 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     2821 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     2881 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     2941 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     3001 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     3061 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     3121 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     3181 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     3241 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     3301 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     3361 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     3421 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     3481 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     3541 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     3601 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     3661 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     3721 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     3781 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     3841 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     3901 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     3961 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     4021 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     4081 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     4141 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     4201 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     4261 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     4321 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     4381 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc
     4441 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     4501 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     4561 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     4621 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     4681 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     4741 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     4801 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     4861 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     4921 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     4981 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     5041 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     5101 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     5161 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     5221 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     5281 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     5341 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     5401 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     5461 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     5521 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     5581 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     5641 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     5701 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     5761 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     5821 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     5881 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     5941 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     6001 ctctctcagc tcggtggaga cgctagagtg agcaagggcg aggagctgtt caccggggtg
     6061 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     6121 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     6181 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     6241 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     6301 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     6361 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     6421 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     6481 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     6541 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     6601 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     6661 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     6721 ggcatggacg agctgtacaa gtctagcagg gctgacccca agaagaagag gaaggtgtga
     6781 atcgatggta cctatgggca ccaaagaacc tgtaaacgtt atctttttaa attgaatgtg
     6841 cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6901 aaaaaaaaaa aaaaaaaaaa aaaactcgag gtagcatgca agtgtacgga attcactcct
     6961 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     7021 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     7081 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     7141 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     7201 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     7261 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     7321 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     7381 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     7441 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt
     7501 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7561 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7621 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     7681 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     7741 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     7801 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     7861 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     7921 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7981 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     8041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     8401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     8461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     8521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     8581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     8641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     8701 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     8761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     8821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     8881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     9001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     9061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     9121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     9181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     9241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     9301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     9361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     9421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     9481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     9541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     9601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     9661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     9721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     9781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     9841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9961 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
