LOCUS       sequence-204223-       10219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-dHDAC1 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10219)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10219)
  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            "Unpublished"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-HDAC1-H141A-Hygro
FEATURES             Location/Qualifiers
     source          1..10219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     195..218
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        258..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        638..841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     792..812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     883..902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        883..901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      909..918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(915..4316,4317..4319,4320..4916,4917,4918..4920,
                     4921..4922,4923..4925,4926..5018)
                     /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(915..941,942..944,945..3431,3432..3434,3435..5018)
                     /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             5025..5045
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5052..5117
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6300..6317)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6306..6530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6601..6617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6601..6617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6614..6636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6625..6641
                     /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(6649..6655,6656..6673,6674..6679))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6694..6715
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6832..6849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7003..7591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7083..7102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7762..8553,8554..8622))
                     /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     8385..8404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8623..8727)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8778..8899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8789..8808)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8843..8862
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8930..9955)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8930..9955)
                     /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(9973..10219,1..253))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10021..10040)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 agactagtga gacgtgctac ttccatttgt cacgtcctgc acgacgcgag ctgcggggcg
       61 ggggggaact tcctgactag gggaggagta gaaggtggcg cgaaggggcc accaaagaac
      121 ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc gaggccagag gccacttgtg
      181 tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc
      241 gcctccccta cccggttgac attgattatt gactagttat taatagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      841 tggtttagtg aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag
      901 ggagagccgc caccatggat aaaaagtatt ctattggttt agccatcggc actaattccg
      961 ttggatgggc tgtcataacc gatgaataca aagtaccttc aaagaaattt aaggtgttgg
     1021 ggaacacaga ccgtcattcg attaaaaaga atcttatcgg tgccctccta ttcgatagtg
     1081 gcgaaacggc agaggcgact cgcctgaaac gaaccgctcg gagaaggtat acacgtcgca
     1141 agaaccgaat atgttactta caagaaattt ttagcaatga gatggccaaa gttgacgatt
     1201 ctttctttca ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa catgaacggc
     1261 accccatctt tggaaacata gtagatgagg tggcatatca tgaaaagtac ccaacgattt
     1321 atcacctcag aaaaaagcta gttgactcaa ctgataaagc ggacctgagg ttaatctact
     1381 tggctcttgc ccatatgata aagttccgtg ggcactttct cattgagggt gatctaaatc
     1441 cggacaactc ggatgtcgac aaactgttca tccagttagt acaaacctat aatcagttgt
     1501 ttgaagagaa ccctataaat gcaagtggcg tggatgcgaa ggctattctt agcgcccgcc
     1561 tctctaaatc ccgacggcta gaaaacctga tcgcacaatt acccggagag aagaaaaatg
     1621 ggttgttcgg taaccttata gcgctctcac taggcctgac accaaatttt aagtcgaact
     1681 tcgacttagc tgaagatgcc aaattgcagc ttagtaagga cacgtacgat gacgatctcg
     1741 acaatctact ggcacaaatt ggagatcagt atgcggactt atttttggct gccaaaaacc
     1801 ttagcgatgc aatcctccta tctgacatac tgagagttaa tactgagatt accaaggcgc
     1861 cgttatccgc ttcaatgatc aaaaggtacg atgaacatca ccaagacttg acacttctca
     1921 aggccctagt ccgtcagcaa ctgcctgaga aatataagga aatattcttt gatcagtcga
     1981 aaaacgggta cgcaggttat attgacggcg gagcgagtca agaggaattc tacaagttta
     2041 tcaaacccat attagagaag atggatggga cggaagagtt gcttgtaaaa ctcaatcgcg
     2101 aagatctact gcgaaagcag cggactttcg acaacggtag cattccacat caaatccact
     2161 taggcgaatt gcatgctata cttagaaggc aggaggattt ttatccgttc ctcaaagaca
     2221 atcgtgaaaa gattgagaaa atcctaacct ttcgcatacc ttactatgtg ggacccctgg
     2281 cccgagggaa ctctcggttc gcatggatga caagaaagtc cgaagaaacg attactccat
     2341 ggaattttga ggaagttgtc gataaaggtg cgtcagctca atcgttcatc gagaggatga
     2401 ccaactttga caagaattta ccgaacgaaa aagtattgcc taagcacagt ttactttacg
     2461 agtatttcac agtgtacaat gaactcacga aagttaagta tgtcactgag ggcatgcgta
     2521 aacccgcctt tctaagcgga gaacagaaga aagcaatagt agatctgtta ttcaagacca
     2581 accgcaaagt gacagttaag caattgaaag aggactactt taagaaaatt gaatgcttcg
     2641 attctgtcga gatctccggg gtagaagatc gatttaatgc gtcacttggt acgtatcatg
     2701 acctcctaaa gataattaaa gataaggact tcctggataa cgaagagaat gaagatatct
     2761 tagaagatat agtgttgact cttaccctct ttgaagatcg ggaaatgatt gaggaaagac
     2821 taaaaacata cgctcacctg ttcgacgata aggttatgaa acagttaaag aggcgtcgct
     2881 atacgggctg gggacgattg tcgcggaaac ttatcaacgg gataagagac aagcaaagtg
     2941 gtaaaactat tctcgatttt ctaaagagcg acggcttcgc caataggaac tttatgcagc
     3001 tgatccatga tgactcttta accttcaaag aggatataca aaaggcacag gtttccggac
     3061 aaggggactc attgcacgaa catattgcga atcttgctgg ttcgccagcc atcaaaaagg
     3121 gcatactcca gacagtcaaa gtagtggatg agctagttaa ggtcatggga cgtcacaaac
     3181 cggaaaacat tgtaatcgag atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa
     3241 acagtcgaga gcggatgaag agaatagaag agggtattaa agaactgggc agccagatct
     3301 taaaggagca tcctgtggaa aatacccaat tgcagaacga gaaactttac ctctattacc
     3361 tacaaaatgg aagggacatg tatgttgatc aggaactgga cataaaccgt ttatctgatt
     3421 acgacgtcga tgccattgta ccccaatcct ttttgaagga cgattcaatc gacaataaag
     3481 tgcttacacg ctcggataag aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg
     3541 taaagaaaat gaagaactat tggcggcagc tcctaaatgc gaaactgata acgcaaagaa
     3601 agttcgataa cttaactaaa gctgagaggg gtggcttgtc tgaacttgac aaggccggat
     3661 ttattaaacg tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca cagatactag
     3721 attcccgaat gaatacgaaa tacgacgaga acgataagct gattcgggaa gtcaaagtaa
     3781 tcactttaaa gtcaaaattg gtgtcggact tcagaaagga ttttcaattc tataaagtta
     3841 gggagataaa taactaccac catgcgcacg acgcttatct taatgccgtc gtagggaccg
     3901 cactcattaa gaaatacccg aagctagaaa gtgagtttgt gtatggtgat tacaaagttt
     3961 atgacgtccg taagatgatc gcgaaaagcg aacaggagat aggcaaggct acagccaaat
     4021 acttctttta ttctaacatt atgaatttct ttaagacgga aatcactctg gcaaacggag
     4081 agatacgcaa acgaccttta attgaaacca atggggagac aggtgaaatc gtatgggata
     4141 agggccggga cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc aacatagtaa
     4201 agaaaactga ggtgcagacc ggagggtttt caaaggaatc gattcttcca aaaaggaata
     4261 gtgataagct catcgctcgt aaaaaggact gggacccgaa aaagtacggt ggcttcgata
     4321 gccctacagt tgcctattct gtcctagtag tggcaaaagt tgagaaggga aaatccaaga
     4381 aactgaagtc agtcaaagaa ttattgggga taacgattat ggagcgctcg tcttttgaaa
     4441 agaaccccat cgacttcctt gaggcgaaag gttacaagga agtaaaaaag gatctcataa
     4501 ttaaactacc aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta
     4561 gcgccggaga gcttcaaaag gggaacgaac tcgcactacc gtctaaatac gtgaatttcc
     4621 tgtatttagc gtcccattac gagaagttga aaggttcacc tgaagataac gaacagaagc
     4681 aactttttgt tgagcagcac aaacattatc tcgacgaaat catagagcaa atttcggaat
     4741 tcagtaagag agtcatccta gctgatgcca atctggacaa agtattaagc gcatacaaca
     4801 agcacaggga taaacccata cgtgagcagg cggaaaatat tatccatttg tttactctta
     4861 ccaacctcgg cgctccagcc gcattcaagt attttgacac aacgatagat cgcaaacgat
     4921 acacttctac caaggaggtg ctagacgcga cactgattca ccaatccatc acgggattat
     4981 atgaaactcg gatagatttg tcacagcttg ggggtgacgg atcccccaag aagaagagga
     5041 aagtctcgag cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca
     5101 aggatgacga tgacaaggct gcaggaggcg gaggtagcac gcagggcacc cggaggaaag
     5161 tctgttacta ctacgacggg gatgttggaa attactatta tggacaaggc cacccaatga
     5221 agcctcaccg aatccgcatg actcataatt tgctgctcaa ctatggtctc taccgaaaaa
     5281 tggaaatcta tcgccctcac aaagccaatg ctgaggagat gaccaagtac cacagcgatg
     5341 actacattaa attcttgcgc tccatccgtc cagataacat gtcggagtac agcaagcaga
     5401 tgcagagatt caacgttggt gaggactgtc cagtattcga tggcctgttt gagttctgtc
     5461 agttgtctac tggtggttct gtggcaagtg ctgtgaaact taataagcag cagacggaca
     5521 tcgctgtgaa ttgggctggg ggcctgcatg ctgcaaagaa gtccgaggca tctggcttct
     5581 gttacgtcaa tgatatcgtc ttggccatcc tggaactgct aaagtatcac cagagggtgc
     5641 tgtacattga cattgatatt caccatggtg acggcgtgga agaggccttc tacaccacgg
     5701 accgggtcat gactgtgtcc tttcataagt atggagagta cttcccagga actggggacc
     5761 tacgggatat cggggctggc aaaggcaagt attatgctgt taactacccg ctccgagacg
     5821 ggattgatga cgagtcctat gaggccattt tcaagccggt catgtccaaa gtaatggaga
     5881 tgttccagcc tagtgcggtg gtcttacagt gtggctcaga ctccctatct ggggatcggt
     5941 taggttgctt caatctaact atcaaaggac acgccaagtg tgtggaattt gtcaagagct
     6001 ttaacctgcc tatgctgatg ctgggaggcg gtggttacac cattcgtaac gttgcccggt
     6061 gctggacata tgagacagct gtggccctgg atacggagat ccctaatgag cttccataca
     6121 atgactactt tgaatacttt ggaccagatt tcaagctcca catcagtcct tccaatatga
     6181 ctaaccagaa cacgaatgag tacctggaga agatcaaaca gcgactgttt gagaacctta
     6241 gaatgctgcc gcacgcacct ggggtccaaa tgcaggcgat ttgaaaaccc gctgatcagc
     6301 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6361 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     6421 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     6481 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     6541 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     6601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6661 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     6721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8761 cgggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     8821 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     8881 aagctgcaat aaacaagttg gggtgggcga agaactctcg gcatctactc tattcctttg
     8941 ccctcggacg agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat
     9001 cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg
     9061 atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa
     9121 ccaagctctg atagagttgg tcaagaccaa tgcggagcat atacgcccgg agccgcggcg
     9181 atcctgcaag ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca
     9241 accacggcct ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct
     9301 cgctccagtc aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga
     9361 aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat
     9421 cgagagcctg cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca
     9481 catggggatc agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt
     9541 gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca
     9601 tggcctccgc gaccggctgc agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg
     9661 tgacaccctg tgcacggcgg gagatgcaat aggtcaggct ctcgctgaat tccccaatgt
     9721 caagcacttc cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt
     9781 tgtagaaacc atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga
     9841 agctgaaagc acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga
     9901 acttttcgat cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatggtgg
     9961 cgggatgcag gtcgaaaggc ccggagatga ggaagaggag aacagcgcgg cagacgtgcg
    10021 cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt cgggcgcccg ccccgcccct
    10081 gagcccgccc ctgagcccgc ccccggaccc accccttccc agcctctgag cccagaaagc
    10141 gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct acccgcttcc attgctcagc
    10201 ggtgctgtcc atctgcacg
//
