LOCUS       sequence-410553-       11727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing both SpCas9 and SaCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11727)
  AUTHORS   Kim J, Kratz AF, Chen S, Sheng J, Kim HK, Zhang L, Singh BK, Chavez 
            A
  TITLE     High-throughput tagging of endogenous loci for rapid 
            characterization of protein function.
  JOURNAL   Sci Adv. 2024 May 3;10(18):eadg8771. doi: 10.1126/sciadv.adg8771. 
            Epub 2024 May 1.
  PUBMED    38691600
REFERENCE   2  (bases 1 to 11727)
  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: "Sci Adv."; 
            date: "2024-05-3"; volume: "10(18):eadg8771. doi"; pages: " 
            10.1126/sciadv.adg8771. Epub 2024 May 1"
COMMENT     Sequence Label: pCAS
FEATURES             Location/Qualifiers
     source          1..11727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        220..238
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     220..237
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             288..335
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             354..374
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             381..4481
                     /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             381..4481
                     /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             381..4481
                     /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             4500..4520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4530..4586
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4599..4625
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             4668..7823
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             7830..7877
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7896..7922
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     polyA_signal    7964..8098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7988..8007)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8042..8061
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8211..8231)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8211..8229)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8250..8266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8250..8266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8263..8285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8274..8290
                     /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(8298..8304,8305..8322,8323..8328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8343..8364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8481..8498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8652..9240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8732..8751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9411..10202,10203..10271))
                     /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     10034..10053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10272..10376)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10402..10857)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10539..10560)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10751..10770
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(10928..11727,1..185)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
       61 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
      121 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
      181 ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat
      241 acaagctact tgttcttttt gcaggatccc atcgattcga attcatgaaa aggccggcgg
      301 ccacgaaaaa ggccggccag gcaaaaaaga aaaagctgga cggcagcgga agccccaaga
      361 agaagaggaa ggtaccatgg gacaagaagt actccattgg gctcgatatc ggtaccaaca
      421 gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
      481 tgggcaatac cgatcgccac agcataaaga agaacctcat tggagccctc ctgttcgact
      541 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
      601 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
      661 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
      721 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
      781 tatatcatct gaggaagaag ctggtagaca gtactgataa ggctgacttg cggttgatct
      841 atctcgcgct ggcgcacatg atcaaatttc ggggacactt cctcatcgag ggggacctga
      901 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
      961 ttttcgagga gaacccgatc aacgcatccg gcgttgacgc caaagcaatc ctgagcgcta
     1021 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     1081 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     1141 acttcgacct ggccgaagat gccaagctgc aactgagcaa agacacctac gatgatgatc
     1201 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     1261 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     1321 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     1381 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     1441 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     1501 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagctgaaca
     1561 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     1621 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     1681 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     1741 tcgctcgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     1801 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     1861 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     1921 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     1981 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     2041 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     2101 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     2161 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     2221 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     2281 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagagacgcc
     2341 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tggcatccga gacaagcaga
     2401 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     2461 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     2521 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     2581 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     2641 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     2701 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     2761 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     2821 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaac cggttgtccg
     2881 actacgacgt ggatcatatc gtgccccaaa gctttctcaa agatgattct attgataata
     2941 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     3001 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     3061 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     3121 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     3181 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     3241 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     3301 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     3361 ctgcacttat caaaaaatat cccaagctgg aatctgaatt tgtttacgga gactataaag
     3421 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     3481 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     3541 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     3601 acaagggtag ggatttcgcg acagtccgca aggtcctgtc catgccgcag gtgaacatcg
     3661 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     3721 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     3781 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     3841 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tccagcttcg
     3901 agaaaaaccc catcgacttt ctcgaagcga aaggatataa agaggtcaaa aaagacctca
     3961 tcattaagct gcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     4021 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     4081 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     4141 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     4201 agttctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     4261 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     4321 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     4381 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     4441 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cgctagcggc accggcgggc
     4501 ccaagaagaa gaggaaggta ggatccggag ccacgaactt ctctctgtta aagcaagcag
     4561 gagatgttga agaaaacccc gggcctggat ccgccgctcc ggcagctaag aaaaagaaac
     4621 tggatttcga atccggaact agtggtatcc acggagtccc agcagccaag cggaactaca
     4681 tcctgggcct ggacatcggc atcaccagcg tgggctacgg catcatcgac tacgagacac
     4741 gggacgtgat cgatgccggc gtgcggctgt tcaaagaggc caacgtggaa aacaacgagg
     4801 gcaggcggag caagagaggc gccagaaggc tgaagcggcg gaggcggcat agaatccaga
     4861 gagtgaagaa gctgctgttc gactacaacc tgctgaccga ccacagcgag ctgagcggca
     4921 tcaaccccta cgaggccaga gtgaagggcc tgagccagaa gctgagcgag gaagagttct
     4981 ctgccgccct gctgcacctg gccaagagaa gaggcgtgca caacgtgaac gaggtggaag
     5041 aggacaccgg caacgagctg tccaccaaag agcagatcag ccggaacagc aaggccctgg
     5101 aagagaaata cgtggccgaa ctgcagctgg aacggctgaa gaaagacggc gaagtgcggg
     5161 gcagcatcaa cagattcaag accagcgact acgtgaaaga agccaaacag ctgctgaagg
     5221 tgcagaaggc ctaccaccag ctggaccaga gcttcatcga cacctacatc gacctgctgg
     5281 aaacccggcg gacctactat gagggacctg gcgagggcag ccccttcggc tggaaggaca
     5341 tcaaagaatg gtacgagatg ctgatgggcc actgcaccta cttccccgag gaactgcgga
     5401 gcgtgaagta cgcctacaac gccgacctgt acaacgccct gaacgacctg aacaatctcg
     5461 tgatcaccag ggacgagaac gagaagctgg aatattacga gaagttccag atcatcgaga
     5521 acgtgttcaa gcagaagaag aagcccaccc tgaagcagat cgccaaagaa atcctcgtga
     5581 acgaagagga tattaagggc tacagagtga ccagcaccgg caagcccgag ttcaccaacc
     5641 tgaaggtgta ccacgacatc aaggacatta ccgcccggaa agagattatt gagaacgccg
     5701 agctgctgga tcagattgcc aagatcctga ccatctacca gagcagcgag gacatccagg
     5761 aagaactgac caatctgaac tccgagctga cccaggaaga gatcgagcag atctctaatc
     5821 tgaagggcta taccggcacc cacaacctga gcctgaaggc catcaacctg atcctggacg
     5881 agctgtggca caccaacgac aaccagatcg ctatcttcaa ccggctgaag ctggtgccca
     5941 agaaggtgga cctgtcccag cagaaagaga tccccaccac cctggtggac gacttcatcc
     6001 tgagccccgt cgtgaagaga agcttcatcc agagcatcaa agtgatcaac gccatcatca
     6061 agaagtacgg cctgcccaac gacatcatta tcgagctggc ccgcgagaag aactccaagg
     6121 acgcccagaa aatgatcaac gagatgcaga agcggaaccg gcagaccaac gagcggatcg
     6181 aggaaatcat ccggaccacc ggcaaagaga acgccaagta cctgatcgag aagatcaagc
     6241 tgcacgacat gcaggaaggc aagtgcctgt acagcctgga agccatccct ctggaagatc
     6301 tgctgaacaa ccccttcaac tatgaggtgg accacatcat ccccagaagc gtgtccttcg
     6361 acaacagctt caacaacaag gtgctcgtga agcaggaaga aaacagcaag aagggcaacc
     6421 ggaccccatt ccagtacctg agcagcagcg acagcaagat cagctacgaa accttcaaga
     6481 agcacatcct gaatctggcc aagggcaagg gcagaatcag caagaccaag aaagagtatc
     6541 tgctggaaga acgggacatc aacaggttct ccgtgcagaa agacttcatc aaccggaacc
     6601 tggtggatac cagatacgcc accagaggcc tgatgaacct gctgcggagc tacttcagag
     6661 tgaacaacct ggacgtgaaa gtgaagtcca tcaatggcgg cttcaccagc tttctgcggc
     6721 ggaagtggaa gtttaagaaa gagcggaaca aggggtacaa gcaccacgcc gaggacgccc
     6781 tgatcattgc caacgccgat ttcatcttca aagagtggaa gaaactggac aaggccaaaa
     6841 aagtgatgga aaaccagatg ttcgaggaaa agcaggccga gagcatgccc gagatcgaaa
     6901 ccgagcagga gtacaaagag atcttcatca ccccccacca gatcaagcac attaaggact
     6961 tcaaggacta caagtacagc caccgggtgg acaagaagcc taatagagag ctgattaacg
     7021 acaccctgta ctccacccgg aaggacgaca agggcaacac cctgatcgtg aacaatctga
     7081 acggcctgta cgacaaggac aatgacaagc tgaaaaagct gatcaacaag agccccgaaa
     7141 agctgctgat gtaccaccac gacccccaga cctaccagaa actgaagctg attatggaac
     7201 agtacggcga cgagaagaat cccctgtaca agtactacga ggaaaccggg aactacctga
     7261 ccaagtactc caaaaaggac aacggccccg tgatcaagaa gattaagtat tacggcaaca
     7321 aactgaacgc ccatctggac atcaccgacg actaccccaa cagcagaaac aaggtcgtga
     7381 agctgtccct gaagccctac agattcgacg tgtacctgga caatggcgtg tacaagttcg
     7441 tgaccgtgaa gaatctggat gtgatcaaaa aagaaaacta ctacgaagtg aatagcaagt
     7501 gctatgagga agctaagaag ctgaagaaga tcagcaacca ggccgagttt atcgcctcct
     7561 tctacaacaa cgatctgatc aagatcaacg gcgagctgta tagagtgatc ggcgtgaaca
     7621 acgacctgct gaaccggatc gaagtgaaca tgatcgacat cacctaccgc gagtacctgg
     7681 aaaacatgaa cgacaagagg ccccccagga tcattaagac aatcgcctcc aagacccaga
     7741 gcattaagaa gtacagcaca gacattctgg gcaacctgta tgaagtgaaa tctaagaagc
     7801 accctcagat catcaaaaag ggccccggga aaaggccggc ggccacgaaa aaggccggcc
     7861 aggcaaaaaa gaaaaagctg gacggcagcg gaagcccggc agctaagaaa aagaaactgg
     7921 atctcgagta ataatctaga actatagtga gtcgtattac gtagatccag acatgataag
     7981 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     8041 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     8101 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     8161 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     8221 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     8281 ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc
     8341 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     8401 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     8461 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     8521 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8581 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     9001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     9061 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     9121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     9181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     9241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     9301 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    10021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    10081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    10141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    10201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    10261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10381 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
    10441 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
    10501 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
    10561 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
    10621 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
    10681 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
    10741 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
    10801 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
    10861 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
    10921 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
    10981 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
    11041 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
    11101 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
    11161 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
    11221 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
    11281 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
    11341 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
    11401 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
    11461 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
    11521 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
    11581 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
    11641 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
    11701 ggcaagtaca ttactattgg aagtacg
//
