LOCUS       sequence-204449-       10021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-SET7 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10021)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10021)
  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            "Unpublished"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-SET7-Hygro
FEATURES             Location/Qualifiers
     source          1..10021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     204..227
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        267..646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        647..850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     801..821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     892..911
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        892..910
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(924..4325,4326..4328,4329..4925,4926,4927..4929,
                     4930..4931,4932..4934,4935..5027)
                     /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(924..950,951..953,954..3440,3441..3443,3444..5027)
                     /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             5034..5054
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5061..5126
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6111..6128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6117..6341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6412..6428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6412..6428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6425..6447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6436..6452
                     /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(6460..6466,6467..6484,6485..6490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6505..6526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6643..6660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6814..7402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6894..6913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7573..8364,8365..8433))
                     /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     8196..8215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8434..8538)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8589..8710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8600..8619)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8654..8673
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8741..9766)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8741..9766)
                     /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(9784..10021,1..262))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9832..9851)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 atctgcacga gactagtgag acgtgctact tccatttgtc acgtcctgca cgacgcgagc
       61 tgcggggcgg gggggaactt cctgactagg ggaggagtag aaggtggcgc gaaggggcca
      121 ccaaagaacg gagccggttg gcgcctaccg gtggatgtgg aatgtgtgcg aggccagagg
      181 ccacttgtgt agcgccaagt gcccagcggg gctgctaaag cgcatgctcc agactgcctt
      241 gggaaaagcg cctcccctac ccggttgaca ttgattattg acttgttatt aatagtaatc
      301 aattacgggg tcattagttc atagcccata tgtggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct ggtttagtga accgtcagat ccgctagaga tccgcggccg ctaatacgac
      901 tcactatagg gagagccgcc accatggata aaaagtattc tattggttta gccatcggca
      961 ctaattccgt tggatgggct gtcataaccg atgaatacaa agtaccttca aagaaattta
     1021 aggtgttggg gaacacagac cgtcattcga ttaaaaagaa tcttatcggt gccctcctat
     1081 tcgatagtgg cgaaacggca gaggcgactc gcctgaaacg aaccgctcgg agaaggtata
     1141 cacgtcgcaa gaaccgaata tgttacttac aagaaatttt tagcaatgag atggccaaag
     1201 ttgacgattc tttctttcac cgtttggaag agtccttcct tgtcgaagag gacaagaaac
     1261 atgaacggca ccccatcttt ggaaacatag tagatgaggt ggcatatcat gaaaagtacc
     1321 caacgattta tcacctcaga aaaaagctag ttgactcaac tgataaagcg gacctgaggt
     1381 taatctactt ggctcttgcc catatgataa agttccgtgg gcactttctc attgagggtg
     1441 atctaaatcc ggacaactcg gatgtcgaca aactgttcat ccagttagta caaacctata
     1501 atcagttgtt tgaagagaac cctataaatg caagtggcgt ggatgcgaag gctattctta
     1561 gcgcccgcct ctctaaatcc cgacggctag aaaacctgat cgcacaatta cccggagaga
     1621 agaaaaatgg gttgttcggt aaccttatag cgctctcact aggcctgaca ccaaatttta
     1681 agtcgaactt cgacttagct gaagatgcca aattgcagct tagtaaggac acgtacgatg
     1741 acgatctcga caatctactg gcacaaattg gagatcagta tgcggactta tttttggctg
     1801 ccaaaaacct tagcgatgca atcctcctat ctgacatact gagagttaat actgagatta
     1861 ccaaggcgcc gttatccgct tcaatgatca aaaggtacga tgaacatcac caagacttga
     1921 cacttctcaa ggccctagtc cgtcagcaac tgcctgagaa atataaggaa atattctttg
     1981 atcagtcgaa aaacgggtac gcaggttata ttgacggcgg agcgagtcaa gaggaattct
     2041 acaagtttat caaacccata ttagagaaga tggatgggac ggaagagttg cttgtaaaac
     2101 tcaatcgcga agatctactg cgaaagcagc ggactttcga caacggtagc attccacatc
     2161 aaatccactt aggcgaattg catgctatac ttagaaggca ggaggatttt tatccgttcc
     2221 tcaaagacaa tcgtgaaaag attgagaaaa tcctaacctt tcgcatacct tactatgtgg
     2281 gacccctggc ccgagggaac tctcggttcg catggatgac aagaaagtcc gaagaaacga
     2341 ttactccatg gaattttgag gaagttgtcg ataaaggtgc gtcagctcaa tcgttcatcg
     2401 agaggatgac caactttgac aagaatttac cgaacgaaaa agtattgcct aagcacagtt
     2461 tactttacga gtatttcaca gtgtacaatg aactcacgaa agttaagtat gtcactgagg
     2521 gcatgcgtaa acccgccttt ctaagcggag aacagaagaa agcaatagta gatctgttat
     2581 tcaagaccaa ccgcaaagtg acagttaagc aattgaaaga ggactacttt aagaaaattg
     2641 aatgcttcga ttctgtcgag atctccgggg tagaagatcg atttaatgcg tcacttggta
     2701 cgtatcatga cctcctaaag ataattaaag ataaggactt cctggataac gaagagaatg
     2761 aagatatctt agaagatata gtgttgactc ttaccctctt tgaagatcgg gaaatgattg
     2821 aggaaagact aaaaacatac gctcacctgt tcgacgataa ggttatgaaa cagttaaaga
     2881 ggcgtcgcta tacgggctgg ggacgattgt cgcggaaact tatcaacggg ataagagaca
     2941 agcaaagtgg taaaactatt ctcgattttc taaagagcga cggcttcgcc aataggaact
     3001 ttatgcagct gatccatgat gactctttaa ccttcaaaga ggatatacaa aaggcacagg
     3061 tttccggaca aggggactca ttgcacgaac atattgcgaa tcttgctggt tcgccagcca
     3121 tcaaaaaggg catactccag acagtcaaag tagtggatga gctagttaag gtcatgggac
     3181 gtcacaaacc ggaaaacatt gtaatcgaga tggcacgcga aaatcaaacg actcagaagg
     3241 ggcaaaaaaa cagtcgagag cggatgaaga gaatagaaga gggtattaaa gaactgggca
     3301 gccagatctt aaaggagcat cctgtggaaa atacccaatt gcagaacgag aaactttacc
     3361 tctattacct acaaaatgga agggacatgt atgttgatca ggaactggac ataaaccgtt
     3421 tatctgatta cgacgtcgat gccattgtac cccaatcctt tttgaaggac gattcaatcg
     3481 acaataaagt gcttacacgc tcggataaga accgagggaa aagtgacaat gttccaagcg
     3541 aggaagtcgt aaagaaaatg aagaactatt ggcggcagct cctaaatgcg aaactgataa
     3601 cgcaaagaaa gttcgataac ttaactaaag ctgagagggg tggcttgtct gaacttgaca
     3661 aggccggatt tattaaacgt cagctcgtgg aaacccgcca aatcacaaag catgttgcac
     3721 agatactaga ttcccgaatg aatacgaaat acgacgagaa cgataagctg attcgggaag
     3781 tcaaagtaat cactttaaag tcaaaattgg tgtcggactt cagaaaggat tttcaattct
     3841 ataaagttag ggagataaat aactaccacc atgcgcacga cgcttatctt aatgccgtcg
     3901 tagggaccgc actcattaag aaatacccga agctagaaag tgagtttgtg tatggtgatt
     3961 acaaagttta tgacgtccgt aagatgatcg cgaaaagcga acaggagata ggcaaggcta
     4021 cagccaaata cttcttttat tctaacatta tgaatttctt taagacggaa atcactctgg
     4081 caaacggaga gatacgcaaa cgacctttaa ttgaaaccaa tggggagaca ggtgaaatcg
     4141 tatgggataa gggccgggac ttcgcgacgg tgagaaaagt tttgtccatg ccccaagtca
     4201 acatagtaaa gaaaactgag gtgcagaccg gagggttttc aaaggaatcg attcttccaa
     4261 aaaggaatag tgataagctc atcgctcgta aaaaggactg ggacccgaaa aagtacggtg
     4321 gcttcgatag ccctacagtt gcctattctg tcctagtagt ggcaaaagtt gagaagggaa
     4381 aatccaagaa actgaagtca gtcaaagaat tattggggat aacgattatg gagcgctcgt
     4441 cttttgaaaa gaaccccatc gacttccttg aggcgaaagg ttacaaggaa gtaaaaaagg
     4501 atctcataat taaactacca aagtatagtc tgtttgagtt agaaaatggc cgaaaacgga
     4561 tgttggctag cgccggagag cttcaaaagg ggaacgaact cgcactaccg tctaaatacg
     4621 tgaatttcct gtatttagcg tcccattacg agaagttgaa aggttcacct gaagataacg
     4681 aacagaagca actttttgtt gagcagcaca aacattatct cgacgaaatc atagagcaaa
     4741 tttcggaatt cagtaagaga gtcatcctag ctgatgccaa tctggacaaa gtattaagcg
     4801 catacaacaa gcacagggat aaacccatac gtgagcaggc ggaaaatatt atccatttgt
     4861 ttactcttac caacctcggc gctccagccg cattcaagta ttttgacaca acgatagatc
     4921 gcaaacgata cacttctacc aaggaggtgc tagacgcgac actgattcac caatccatca
     4981 cgggattata tgaaactcgg atagatttgt cacagcttgg gggtgacgga tcccccaaga
     5041 agaagaggaa agtctcgagc gactacaaag accatgacgg tgattataaa gatcatgaca
     5101 tcgattacaa ggatgacgat gacaaggctg caggaggcgg aggtagcttc ttctttgatg
     5161 gcagcaccct ggaggggtat tatgtggatg atgccttgca gggccaggga gtttacactt
     5221 acgaagatgg gggagttctc cagggcacgt atgtagacgg agagctgaac ggtccagccc
     5281 aggaatatga cacagatggg agactgatct tcaaggggca gtataaagat aacattcgtc
     5341 atggagtgtg ctggatatat tacccagatg gaggaagcct tgtaggagaa gtaaatgaag
     5401 atggggagat gactggagag aagatagcct atgtgtaccc tgatgagagg accgcacttt
     5461 atgggaaatt tattgatgga gagatgatag aaggcaaact ggctaccctt atgtccactg
     5521 aagaagggag gcctcacttt gaactgatgc ctggaaattc agtgtaccac tttgataagt
     5581 cgacttcatc ttgcatttct accaatgctc ttcttccaga tccttatgaa tcagaaaggg
     5641 tttatgttgc tgaatctctt atttccagtg ctggagaagg acttttttca aaggtagctg
     5701 tgggacctaa tactgttatg tctttttata atggagttcg aattacacac caagaggttg
     5761 acagcaggga ctgggccctt aatgggaaca ccctctccct tgatgaagaa acggtcattg
     5821 atgtgcctga gccctataac cacgtatcca agtactgtgc ctccttggga cacaaggcaa
     5881 atcactcctt cactccaaac tgcatctacg atatgtttgt ccacccccgt tttgggccca
     5941 tcaaatgcat ccgcaccctg agagcagtgg aggccgatga agagctcacc gttgcctatg
     6001 gctatgacca cagccccccc gggaagagtg ggcctgaagc ccctgagtgg taccaggtgg
     6061 agctgaaggc cttccaggcc acccagcaaa agtgaaaacc cgctgatcag cctcgactgt
     6121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     6181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     6241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     6301 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     6361 cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6421 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6481 aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6541 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6601 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7561 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7741 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8101 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8401 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acgggtatac
     8581 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     8641 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     8701 taaacaagtt ggggtgggcg aagaactctc ggcatctact ctattccttt gccctcggac
     8761 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     8821 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     8881 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     8941 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa
     9001 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     9061 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     9121 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     9181 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     9241 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     9301 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     9361 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg
     9421 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     9481 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt
     9541 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     9601 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     9661 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     9721 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatggtg gcgggatgca
     9781 ggtcgaaagg cccggagatg aggaagagga gaacagcgcg gcagacgtgc gcttttgaag
     9841 cgtgcagaat gccgggcctc cggaggacct tcgggcgccc gccccgcccc tgagcccgcc
     9901 cctgagcccg cccccggacc caccccttcc cagcctctga gcccagaaag cgaaggagca
     9961 aagctgctat tggccgctgc cccaaaggcc tacccgcttc cattgctcag cggtgctgtc
    10021 c
//
