LOCUS       sequence-204469-        9922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-G9a fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9922)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9922)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p-dCas9-G9a-Hygro
FEATURES             Location/Qualifiers
     source          1..9922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..243
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        283..662
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        663..866
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     817..837
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     908..927
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        908..926
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      934..943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(940..4341,4342..4344,4345..4941,4942,4943..4945,
                     4946..4947,4948..4950,4951..5043)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(940..966,967..969,970..3456,3457..3459,3460..5043)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5050..5070
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5077..5142
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6028..6045)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6034..6258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6329..6345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6329..6345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6342..6364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6353..6369
                     /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(6377..6383,6384..6401,6402..6407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6422..6443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6560..6577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6731..7319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6811..6830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7490..8281,8282..8350))
                     /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     8113..8132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8351..8455)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8506..8627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8517..8536)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8571..8590
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8658..9683)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8658..9683)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(join(9701..9922,1..278))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9749..9768)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 ctcagcggtg ctgtccatct gcacgagact agtgagacgt gctacttcca tttgtcacgt
       61 cctgcacgac gcgagctgcg gggcgggggg gaacttcctg actaggggag gagtagaagg
      121 tggcgcgaag gggccaccaa agaacggagc cggttggcgc ctaccggtgg atgtggaatg
      181 tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc agcggggctg ctaaagcgca
      241 tgctccagac tgccttggga aaagcgcctc ccctacccgg ttgacattga ttattgacta
      301 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      361 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      421 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      481 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      541 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      601 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      661 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      721 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      781 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      841 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
      901 cggccgctaa tacgactcac tatagggaga gccgccacca tggataaaaa gtattctatt
      961 ggtttagcca tcggcactaa ttccgttgga tgggctgtca taaccgatga atacaaagta
     1021 ccttcaaaga aatttaaggt gttggggaac acagaccgtc attcgattaa aaagaatctt
     1081 atcggtgccc tcctattcga tagtggcgaa acggcagagg cgactcgcct gaaacgaacc
     1141 gctcggagaa ggtatacacg tcgcaagaac cgaatatgtt acttacaaga aatttttagc
     1201 aatgagatgg ccaaagttga cgattctttc tttcaccgtt tggaagagtc cttccttgtc
     1261 gaagaggaca agaaacatga acggcacccc atctttggaa acatagtaga tgaggtggca
     1321 tatcatgaaa agtacccaac gatttatcac ctcagaaaaa agctagttga ctcaactgat
     1381 aaagcggacc tgaggttaat ctacttggct cttgcccata tgataaagtt ccgtgggcac
     1441 tttctcattg agggtgatct aaatccggac aactcggatg tcgacaaact gttcatccag
     1501 ttagtacaaa cctataatca gttgtttgaa gagaacccta taaatgcaag tggcgtggat
     1561 gcgaaggcta ttcttagcgc ccgcctctct aaatcccgac ggctagaaaa cctgatcgca
     1621 caattacccg gagagaagaa aaatgggttg ttcggtaacc ttatagcgct ctcactaggc
     1681 ctgacaccaa attttaagtc gaacttcgac ttagctgaag atgccaaatt gcagcttagt
     1741 aaggacacgt acgatgacga tctcgacaat ctactggcac aaattggaga tcagtatgcg
     1801 gacttatttt tggctgccaa aaaccttagc gatgcaatcc tcctatctga catactgaga
     1861 gttaatactg agattaccaa ggcgccgtta tccgcttcaa tgatcaaaag gtacgatgaa
     1921 catcaccaag acttgacact tctcaaggcc ctagtccgtc agcaactgcc tgagaaatat
     1981 aaggaaatat tctttgatca gtcgaaaaac gggtacgcag gttatattga cggcggagcg
     2041 agtcaagagg aattctacaa gtttatcaaa cccatattag agaagatgga tgggacggaa
     2101 gagttgcttg taaaactcaa tcgcgaagat ctactgcgaa agcagcggac tttcgacaac
     2161 ggtagcattc cacatcaaat ccacttaggc gaattgcatg ctatacttag aaggcaggag
     2221 gatttttatc cgttcctcaa agacaatcgt gaaaagattg agaaaatcct aacctttcgc
     2281 ataccttact atgtgggacc cctggcccga gggaactctc ggttcgcatg gatgacaaga
     2341 aagtccgaag aaacgattac tccatggaat tttgaggaag ttgtcgataa aggtgcgtca
     2401 gctcaatcgt tcatcgagag gatgaccaac tttgacaaga atttaccgaa cgaaaaagta
     2461 ttgcctaagc acagtttact ttacgagtat ttcacagtgt acaatgaact cacgaaagtt
     2521 aagtatgtca ctgagggcat gcgtaaaccc gcctttctaa gcggagaaca gaagaaagca
     2581 atagtagatc tgttattcaa gaccaaccgc aaagtgacag ttaagcaatt gaaagaggac
     2641 tactttaaga aaattgaatg cttcgattct gtcgagatct ccggggtaga agatcgattt
     2701 aatgcgtcac ttggtacgta tcatgacctc ctaaagataa ttaaagataa ggacttcctg
     2761 gataacgaag agaatgaaga tatcttagaa gatatagtgt tgactcttac cctctttgaa
     2821 gatcgggaaa tgattgagga aagactaaaa acatacgctc acctgttcga cgataaggtt
     2881 atgaaacagt taaagaggcg tcgctatacg ggctggggac gattgtcgcg gaaacttatc
     2941 aacgggataa gagacaagca aagtggtaaa actattctcg attttctaaa gagcgacggc
     3001 ttcgccaata ggaactttat gcagctgatc catgatgact ctttaacctt caaagaggat
     3061 atacaaaagg cacaggtttc cggacaaggg gactcattgc acgaacatat tgcgaatctt
     3121 gctggttcgc cagccatcaa aaagggcata ctccagacag tcaaagtagt ggatgagcta
     3181 gttaaggtca tgggacgtca caaaccggaa aacattgtaa tcgagatggc acgcgaaaat
     3241 caaacgactc agaaggggca aaaaaacagt cgagagcgga tgaagagaat agaagagggt
     3301 attaaagaac tgggcagcca gatcttaaag gagcatcctg tggaaaatac ccaattgcag
     3361 aacgagaaac tttacctcta ttacctacaa aatggaaggg acatgtatgt tgatcaggaa
     3421 ctggacataa accgtttatc tgattacgac gtcgatgcca ttgtacccca atcctttttg
     3481 aaggacgatt caatcgacaa taaagtgctt acacgctcgg ataagaaccg agggaaaagt
     3541 gacaatgttc caagcgagga agtcgtaaag aaaatgaaga actattggcg gcagctccta
     3601 aatgcgaaac tgataacgca aagaaagttc gataacttaa ctaaagctga gaggggtggc
     3661 ttgtctgaac ttgacaaggc cggatttatt aaacgtcagc tcgtggaaac ccgccaaatc
     3721 acaaagcatg ttgcacagat actagattcc cgaatgaata cgaaatacga cgagaacgat
     3781 aagctgattc gggaagtcaa agtaatcact ttaaagtcaa aattggtgtc ggacttcaga
     3841 aaggattttc aattctataa agttagggag ataaataact accaccatgc gcacgacgct
     3901 tatcttaatg ccgtcgtagg gaccgcactc attaagaaat acccgaagct agaaagtgag
     3961 tttgtgtatg gtgattacaa agtttatgac gtccgtaaga tgatcgcgaa aagcgaacag
     4021 gagataggca aggctacagc caaatacttc ttttattcta acattatgaa tttctttaag
     4081 acggaaatca ctctggcaaa cggagagata cgcaaacgac ctttaattga aaccaatggg
     4141 gagacaggtg aaatcgtatg ggataagggc cgggacttcg cgacggtgag aaaagttttg
     4201 tccatgcccc aagtcaacat agtaaagaaa actgaggtgc agaccggagg gttttcaaag
     4261 gaatcgattc ttccaaaaag gaatagtgat aagctcatcg ctcgtaaaaa ggactgggac
     4321 ccgaaaaagt acggtggctt cgatagccct acagttgcct attctgtcct agtagtggca
     4381 aaagttgaga agggaaaatc caagaaactg aagtcagtca aagaattatt ggggataacg
     4441 attatggagc gctcgtcttt tgaaaagaac cccatcgact tccttgaggc gaaaggttac
     4501 aaggaagtaa aaaaggatct cataattaaa ctaccaaagt atagtctgtt tgagttagaa
     4561 aatggccgaa aacggatgtt ggctagcgcc ggagagcttc aaaaggggaa cgaactcgca
     4621 ctaccgtcta aatacgtgaa tttcctgtat ttagcgtccc attacgagaa gttgaaaggt
     4681 tcacctgaag ataacgaaca gaagcaactt tttgttgagc agcacaaaca ttatctcgac
     4741 gaaatcatag agcaaatttc ggaattcagt aagagagtca tcctagctga tgccaatctg
     4801 gacaaagtat taagcgcata caacaagcac agggataaac ccatacgtga gcaggcggaa
     4861 aatattatcc atttgtttac tcttaccaac ctcggcgctc cagccgcatt caagtatttt
     4921 gacacaacga tagatcgcaa acgatacact tctaccaagg aggtgctaga cgcgacactg
     4981 attcaccaat ccatcacggg attatatgaa actcggatag atttgtcaca gcttgggggt
     5041 gacggatccc ccaagaagaa gaggaaagtc tcgagcgact acaaagacca tgacggtgat
     5101 tataaagatc atgacatcga ttacaaggat gacgatgaca aggctgcagg aggcggaggt
     5161 agcggctatg agaacgtgcc cattccctgt gtcaacggtg tggatgggga gccctgccct
     5221 gaggattaca agtacatctc agagaactgc gagacgtcca ccatgaacat cgatcgcaac
     5281 atcacccacc tgcagcactg cacgtgtgtg gacgactgct ctagctccaa ctgcctgtgc
     5341 ggccagctca gcatccggtg ctggtatgac aaggatgggc gattgctcca ggaatttaac
     5401 aagattgagc ctccgctgat tttcgagtgt aaccaggcgt gctcatgctg gagaaactgc
     5461 aagaaccggg tcgtacagag tggcatcaag gtgcggctac agctctaccg aacagccaag
     5521 atgggctggg gggtccgcgc cctgcagacc atcccacagg ggaccttcat ctgcgagtat
     5581 gtcggggagc tgatctctga tgctgaggct gatgtgagag aggatgattc ttacctcttc
     5641 gacttagaca acaaggatgg agaggtgtac tgcatagatg cccgttacta tggcaacatc
     5701 agccgcttca tcaaccacct gtgtgacccc aacatcattc ccgtccgggt cttcatgctg
     5761 caccaagacc tgcgatttcc acgcatcgcc ttcttcagtt cccgagacat ccggactggg
     5821 gaggagctag ggtttgacta tggcgaccgc ttctgggaca tcaaaagcaa atatttcacc
     5881 tgccaatgtg gctctgagaa gtgcaagcac tcagccgaag ccattgccct ggagcagagc
     5941 cgtctggccc gcctggaccc acaccctgag ctgctgcccg agctcggctc cctgccccct
     6001 gtcaacacat gaaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6061 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6121 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6181 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6241 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     6301 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     6421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6481 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6541 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7141 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7801 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8461 ttccccgaaa agtgccacct gacgtcgacg ggtatacaga catgataaga tacattgatg
     8521 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     8581 atgctattgc tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag
     8641 aactctcggc atctactcta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca
     8701 ctatcggcga gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt
     8761 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc
     8821 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg
     8881 cggagcatat acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag
     8941 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc
     9001 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca
     9061 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg
     9121 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg
     9181 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca
     9241 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg
     9301 ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc
     9361 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag
     9421 gtcaggctct cgctgaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat
     9481 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
     9541 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
     9601 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
     9661 gcggtgagtt caggcttttt catggtggcg ggatgcaggt cgaaaggccc ggagatgagg
     9721 aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg
     9781 aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac
     9841 cccttcccag cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc
     9901 aaaggcctac ccgcttccat tg
//
