LOCUS       sequence-472945-        9302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of human codon-optimized Cas9 nuclease and GFP, 
            plasmid optimized for expression in human pluripotent stem cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9302)
  AUTHORS   Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA, Musunuru K
  TITLE     Enhanced efficiency of human pluripotent stem cell genome editing 
            through replacing TALENs with CRISPRs.
  JOURNAL   Cell Stem Cell. 2013 Apr 4;12(4):393-4. doi: 
            10.1016/j.stem.2013.03.006.
  PUBMED    23561441
REFERENCE   2  (bases 1 to 9302)
  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.stem.2013.03"; journalName: "Cell Stem Cell"; date: 
            "2013-04-4- 4"; volume: "12"; issue: "4"; pages: "393-4"
COMMENT     Sequence Label: pCas9_GFP
FEATURES             Location/Qualifiers
     source          1..9302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(79..98)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        270..649
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        651..928
                     /label=chicken beta-actin promoter
     primer_bind     1895..1917
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1979..1998
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2032..2041
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2038..6141
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2038..6141
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2038..6141
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6154..6174
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6190..6243
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6244..6246,6247..6249,6250..6963)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6244..6246,6247..6249,6250..6960)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6289..6310)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6550..6569)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6897..6918
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7006..7127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7043..7062)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7097..7116
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7176..7192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7176..7192)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7189..7211)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7200..7216
                     /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(7224..7230,7231..7248,7249..7254))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7269..7290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7407..7424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7578..8166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7658..7677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8337..9128,9129..9197))
                     /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     8960..8979
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9198..9302)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcgat ctcccgatcc
       61 cctagggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
      121 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
      181 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
      241 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
      301 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      361 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      421 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      481 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      541 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      601 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      661 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      721 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
      781 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
      841 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
      901 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      961 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     1021 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     1081 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     1141 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1201 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     1261 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggttgtcg
     1321 ggcagcagca cggggccgtc gccgatgggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1381 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1441 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1501 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1561 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1621 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1681 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1741 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1801 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1861 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1921 ggctctagcg cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1981 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcgcggc cgccaccatg
     2041 gacaagaagt actccattgg gctcgatatc ggcacaaaca gcgtcggctg ggccgtcatt
     2101 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
     2161 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
     2221 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
     2281 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     2341 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     2401 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     2461 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     2521 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     2581 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     2641 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     2701 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     2761 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     2821 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     2881 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     2941 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     3001 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     3061 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     3121 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     3181 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     3241 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     3301 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     3361 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     3421 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     3481 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     3541 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     3601 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     3661 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     3721 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     3781 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     3841 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     3901 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     3961 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     4021 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     4081 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     4141 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     4201 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     4261 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     4321 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     4381 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     4441 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     4501 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc
     4561 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     4621 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     4681 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     4741 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     4801 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     4861 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     4921 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     4981 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     5041 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     5101 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     5161 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     5221 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     5281 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     5341 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     5401 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     5461 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     5521 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     5581 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     5641 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     5701 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     5761 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     5821 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     5881 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     5941 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     6001 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     6061 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     6121 ctctctcagc tcggtggaga cagcagggct gaccccaaga agaagaggaa ggtgaggtcc
     6181 ggcggcggag agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     6241 ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     6301 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     6361 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     6421 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     6481 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     6541 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     6601 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     6661 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     6721 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     6781 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     6841 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     6901 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     6961 taactgcagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     7021 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     7081 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     7141 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7201 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctag
     7261 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7321 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7381 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7741 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7981 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     8041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8221 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8281 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8341 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8401 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8461 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8521 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8581 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8641 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8701 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8761 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8821 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8881 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8941 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9001 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9061 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9121 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9181 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9241 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9301 cg
//
