LOCUS       sequence-203185-       10021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-dSET7 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  JOURNAL   Unpublished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            "Unpublished"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-H297G-Hygro
FEATURES             Location/Qualifiers
     source          1..10021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     327..350
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        390..769
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        770..973
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     924..944
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1015..1034
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1015..1033
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1041..1050
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1047..4448,4449..4451,4452..5048,5049,5050..5052,
                     5053..5054,5055..5057,5058..5150)
                     /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(1047..1073,1074..1076,1077..3563,3564..3566,
                     3567..5150)
                     /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             5157..5177
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5184..5249
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6234..6251)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6240..6464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6535..6551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6535..6551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6548..6570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6559..6575
                     /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(6583..6589,6590..6607,6608..6613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6628..6649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6766..6783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6937..7525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7017..7036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7696..8487,8488..8556))
                     /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     8319..8338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8557..8661)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8712..8833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8723..8742)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8777..8796
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8864..9889)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8864..9889)
                     /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(9907..10021,1..385))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9955..9974)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 cccctgagcc cgcccccgga cccacccctt cccagcctct gagcccagaa agcgaaggag
       61 caaagctgct attggccgct gccccaaagg cctacccgct tccattgctc agcggtgctg
      121 tccatctgca cgagactagt gagacgtgct acttccattt gtcacgtcct gcacgacgcg
      181 agctgcgggg cgggggggaa cttcctgact aggggaggag tagaaggtgg cgcgaagggg
      241 ccaccaaaga acggagccgg ttggcgccta ccggtggatg tggaatgtgt gcgaggccag
      301 aggccacttg tgtagcgcca agtgcccagc ggggctgcta aagcgcatgc tccagactgc
      361 cttgggaaaa gcgcctcccc tacccggttg acattgatta ttgacttgtt attaatagta
      421 atcaattacg gggtcattag ttcatagccc atatgtggag ttccgcgtta cataacttac
      481 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      541 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      601 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      661 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      721 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      781 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      841 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      901 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      961 tataagcaga gctggtttag tgaaccgtca gatccgctag agatccgcgg ccgctaatac
     1021 gactcactat agggagagcc gccaccatgg ataaaaagta ttctattggt ttagccatcg
     1081 gcactaattc cgttggatgg gctgtcataa ccgatgaata caaagtacct tcaaagaaat
     1141 ttaaggtgtt ggggaacaca gaccgtcatt cgattaaaaa gaatcttatc ggtgccctcc
     1201 tattcgatag tggcgaaacg gcagaggcga ctcgcctgaa acgaaccgct cggagaaggt
     1261 atacacgtcg caagaaccga atatgttact tacaagaaat ttttagcaat gagatggcca
     1321 aagttgacga ttctttcttt caccgtttgg aagagtcctt ccttgtcgaa gaggacaaga
     1381 aacatgaacg gcaccccatc tttggaaaca tagtagatga ggtggcatat catgaaaagt
     1441 acccaacgat ttatcacctc agaaaaaagc tagttgactc aactgataaa gcggacctga
     1501 ggttaatcta cttggctctt gcccatatga taaagttccg tgggcacttt ctcattgagg
     1561 gtgatctaaa tccggacaac tcggatgtcg acaaactgtt catccagtta gtacaaacct
     1621 ataatcagtt gtttgaagag aaccctataa atgcaagtgg cgtggatgcg aaggctattc
     1681 ttagcgcccg cctctctaaa tcccgacggc tagaaaacct gatcgcacaa ttacccggag
     1741 agaagaaaaa tgggttgttc ggtaacctta tagcgctctc actaggcctg acaccaaatt
     1801 ttaagtcgaa cttcgactta gctgaagatg ccaaattgca gcttagtaag gacacgtacg
     1861 atgacgatct cgacaatcta ctggcacaaa ttggagatca gtatgcggac ttatttttgg
     1921 ctgccaaaaa ccttagcgat gcaatcctcc tatctgacat actgagagtt aatactgaga
     1981 ttaccaaggc gccgttatcc gcttcaatga tcaaaaggta cgatgaacat caccaagact
     2041 tgacacttct caaggcccta gtccgtcagc aactgcctga gaaatataag gaaatattct
     2101 ttgatcagtc gaaaaacggg tacgcaggtt atattgacgg cggagcgagt caagaggaat
     2161 tctacaagtt tatcaaaccc atattagaga agatggatgg gacggaagag ttgcttgtaa
     2221 aactcaatcg cgaagatcta ctgcgaaagc agcggacttt cgacaacggt agcattccac
     2281 atcaaatcca cttaggcgaa ttgcatgcta tacttagaag gcaggaggat ttttatccgt
     2341 tcctcaaaga caatcgtgaa aagattgaga aaatcctaac ctttcgcata ccttactatg
     2401 tgggacccct ggcccgaggg aactctcggt tcgcatggat gacaagaaag tccgaagaaa
     2461 cgattactcc atggaatttt gaggaagttg tcgataaagg tgcgtcagct caatcgttca
     2521 tcgagaggat gaccaacttt gacaagaatt taccgaacga aaaagtattg cctaagcaca
     2581 gtttacttta cgagtatttc acagtgtaca atgaactcac gaaagttaag tatgtcactg
     2641 agggcatgcg taaacccgcc tttctaagcg gagaacagaa gaaagcaata gtagatctgt
     2701 tattcaagac caaccgcaaa gtgacagtta agcaattgaa agaggactac tttaagaaaa
     2761 ttgaatgctt cgattctgtc gagatctccg gggtagaaga tcgatttaat gcgtcacttg
     2821 gtacgtatca tgacctccta aagataatta aagataagga cttcctggat aacgaagaga
     2881 atgaagatat cttagaagat atagtgttga ctcttaccct ctttgaagat cgggaaatga
     2941 ttgaggaaag actaaaaaca tacgctcacc tgttcgacga taaggttatg aaacagttaa
     3001 agaggcgtcg ctatacgggc tggggacgat tgtcgcggaa acttatcaac gggataagag
     3061 acaagcaaag tggtaaaact attctcgatt ttctaaagag cgacggcttc gccaatagga
     3121 actttatgca gctgatccat gatgactctt taaccttcaa agaggatata caaaaggcac
     3181 aggtttccgg acaaggggac tcattgcacg aacatattgc gaatcttgct ggttcgccag
     3241 ccatcaaaaa gggcatactc cagacagtca aagtagtgga tgagctagtt aaggtcatgg
     3301 gacgtcacaa accggaaaac attgtaatcg agatggcacg cgaaaatcaa acgactcaga
     3361 aggggcaaaa aaacagtcga gagcggatga agagaataga agagggtatt aaagaactgg
     3421 gcagccagat cttaaaggag catcctgtgg aaaataccca attgcagaac gagaaacttt
     3481 acctctatta cctacaaaat ggaagggaca tgtatgttga tcaggaactg gacataaacc
     3541 gtttatctga ttacgacgtc gatgccattg taccccaatc ctttttgaag gacgattcaa
     3601 tcgacaataa agtgcttaca cgctcggata agaaccgagg gaaaagtgac aatgttccaa
     3661 gcgaggaagt cgtaaagaaa atgaagaact attggcggca gctcctaaat gcgaaactga
     3721 taacgcaaag aaagttcgat aacttaacta aagctgagag gggtggcttg tctgaacttg
     3781 acaaggccgg atttattaaa cgtcagctcg tggaaacccg ccaaatcaca aagcatgttg
     3841 cacagatact agattcccga atgaatacga aatacgacga gaacgataag ctgattcggg
     3901 aagtcaaagt aatcacttta aagtcaaaat tggtgtcgga cttcagaaag gattttcaat
     3961 tctataaagt tagggagata aataactacc accatgcgca cgacgcttat cttaatgccg
     4021 tcgtagggac cgcactcatt aagaaatacc cgaagctaga aagtgagttt gtgtatggtg
     4081 attacaaagt ttatgacgtc cgtaagatga tcgcgaaaag cgaacaggag ataggcaagg
     4141 ctacagccaa atacttcttt tattctaaca ttatgaattt ctttaagacg gaaatcactc
     4201 tggcaaacgg agagatacgc aaacgacctt taattgaaac caatggggag acaggtgaaa
     4261 tcgtatggga taagggccgg gacttcgcga cggtgagaaa agttttgtcc atgccccaag
     4321 tcaacatagt aaagaaaact gaggtgcaga ccggagggtt ttcaaaggaa tcgattcttc
     4381 caaaaaggaa tagtgataag ctcatcgctc gtaaaaagga ctgggacccg aaaaagtacg
     4441 gtggcttcga tagccctaca gttgcctatt ctgtcctagt agtggcaaaa gttgagaagg
     4501 gaaaatccaa gaaactgaag tcagtcaaag aattattggg gataacgatt atggagcgct
     4561 cgtcttttga aaagaacccc atcgacttcc ttgaggcgaa aggttacaag gaagtaaaaa
     4621 aggatctcat aattaaacta ccaaagtata gtctgtttga gttagaaaat ggccgaaaac
     4681 ggatgttggc tagcgccgga gagcttcaaa aggggaacga actcgcacta ccgtctaaat
     4741 acgtgaattt cctgtattta gcgtcccatt acgagaagtt gaaaggttca cctgaagata
     4801 acgaacagaa gcaacttttt gttgagcagc acaaacatta tctcgacgaa atcatagagc
     4861 aaatttcgga attcagtaag agagtcatcc tagctgatgc caatctggac aaagtattaa
     4921 gcgcatacaa caagcacagg gataaaccca tacgtgagca ggcggaaaat attatccatt
     4981 tgtttactct taccaacctc ggcgctccag ccgcattcaa gtattttgac acaacgatag
     5041 atcgcaaacg atacacttct accaaggagg tgctagacgc gacactgatt caccaatcca
     5101 tcacgggatt atatgaaact cggatagatt tgtcacagct tgggggtgac ggatccccca
     5161 agaagaagag gaaagtctcg agcgactaca aagaccatga cggtgattat aaagatcatg
     5221 acatcgatta caaggatgac gatgacaagg ctgcaggagg cggaggtagc ttcttctttg
     5281 atggcagcac cctggagggg tattatgtgg atgatgcctt gcagggccag ggagtttaca
     5341 cttacgaaga tgggggagtt ctccagggca cgtatgtaga cggagagctg aacggtccag
     5401 cccaggaata tgacacagat gggagactga tcttcaaggg gcagtataaa gataacattc
     5461 gtcatggagt gtgctggata tattacccag atggaggaag ccttgtagga gaagtaaatg
     5521 aagatgggga gatgactgga gagaagatag cctatgtgta ccctgatgag aggaccgcac
     5581 tttatgggaa atttattgat ggagagatga tagaaggcaa actggctacc cttatgtcca
     5641 ctgaagaagg gaggcctcac tttgaactga tgcctggaaa ttcagtgtac cactttgata
     5701 agtcgacttc atcttgcatt tctaccaatg ctcttcttcc agatccttat gaatcagaaa
     5761 gggtttatgt tgctgaatct cttatttcca gtgctggaga aggacttttt tcaaaggtag
     5821 ctgtgggacc taatactgtt atgtcttttt ataatggagt tcgaattaca caccaagagg
     5881 ttgacagcag ggactgggcc cttaatggga acaccctctc ccttgatgaa gaaacggtca
     5941 ttgatgtgcc tgagccctat aaccacgtat ccaagtactg tgcctccttg ggacacaagg
     6001 caaatggatc cttcactcca aactgcatct acgatatgtt tgtccacccc cgttttgggc
     6061 ccatcaaatg catccgcacc ctgagagcag tggaggccga tgaagagctc accgttgcct
     6121 atggctatga ccacagcccc cccgggaaga gtgggcctga agcccctgag tggtaccagg
     6181 tggagctgaa ggccttccag gccacccagc aaaagtgaaa acccgctgat cagcctcgac
     6241 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6301 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6361 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6421 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6481 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6541 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6601 cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacgggta
     8701 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     8761 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     8821 caataaacaa gttggggtgg gcgaagaact ctcggcatct actctattcc tttgccctcg
     8881 gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc
     8941 agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct ccggatcgga
     9001 cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc
     9061 tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg
     9121 caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg
     9181 gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc gcctcgctcc
     9241 agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag ccgaaatccg
     9301 cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc tcatcgagag
     9361 cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga tacacatggg
     9421 gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc
     9481 cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca tccatggcct
     9541 ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc aacgtgacac
     9601 cctgtgcacg gcgggagatg caataggtca ggctctcgct gaattcccca atgtcaagca
     9661 cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga tctttgtaga
     9721 aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca tcgaagctga
     9781 aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt
     9841 cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttcatg gtggcgggat
     9901 gcaggtcgaa aggcccggag atgaggaaga ggagaacagc gcggcagacg tgcgcttttg
     9961 aagcgtgcag aatgccgggc ctccggagga ccttcgggcg cccgccccgc ccctgagccc
    10021 g
//
