LOCUS       sequence-204405-       11122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-dLSD1 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11122)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11122)
  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            "Unpublished"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-LSD1-K661A-Hygro
FEATURES             Location/Qualifiers
     source          1..11122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(147..646)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(195..214)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     588..611
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        651..1030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1031..1234
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1185..1205
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1276..1295
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1276..1294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1302..1311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1308..4709,4710..4712,4713..5309,5310,5311..5313,
                     5314..5315,5316..5318,5319..5411)
                     /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(1308..1334,1335..1337,1338..3824,3825..3827,
                     3828..5411)
                     /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             5418..5438
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5445..5510
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7596..7613)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7602..7826
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7897..7913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7897..7913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7910..7932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7921..7937
                     /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(7945..7951,7952..7969,7970..7975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7990..8011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8128..8145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8299..8887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8379..8398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9058..9849,9850..9918))
                     /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     9681..9700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9919..10023)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10074..10195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10085..10104)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10139..10158
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(10226..11122,1..129))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(join(10226..11122,1..129))
                     /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"
ORIGIN
        1 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc
       61 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg
      121 ctttttcatg gtggcgggat gcaggtcgaa aggcccggag atgaggaaga ggagaacagc
      181 gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc ctccggagga ccttcgggcg
      241 cccgccccgc ccctgagccc gcccctgagc ccgcccccgg acccacccct tcccagcctc
      301 tgagcccaga aagcgaagga gcaaagctgc tattggccgc tgccccaaag gcctacccgc
      361 ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc tacttccatt
      421 tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac taggggagga
      481 gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct accggtggat
      541 gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag cggggctgct
      601 aaagcgcatg ctccagactg ccttgggaaa agcgcctccc ctacccggtt gacattgatt
      661 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      721 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      781 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      841 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      901 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      961 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1021 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     1081 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     1141 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     1201 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     1261 gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg gataaaaagt
     1321 attctattgg tttagccatc ggcactaatt ccgttggatg ggctgtcata accgatgaat
     1381 acaaagtacc ttcaaagaaa tttaaggtgt tggggaacac agaccgtcat tcgattaaaa
     1441 agaatcttat cggtgccctc ctattcgata gtggcgaaac ggcagaggcg actcgcctga
     1501 aacgaaccgc tcggagaagg tatacacgtc gcaagaaccg aatatgttac ttacaagaaa
     1561 tttttagcaa tgagatggcc aaagttgacg attctttctt tcaccgtttg gaagagtcct
     1621 tccttgtcga agaggacaag aaacatgaac ggcaccccat ctttggaaac atagtagatg
     1681 aggtggcata tcatgaaaag tacccaacga tttatcacct cagaaaaaag ctagttgact
     1741 caactgataa agcggacctg aggttaatct acttggctct tgcccatatg ataaagttcc
     1801 gtgggcactt tctcattgag ggtgatctaa atccggacaa ctcggatgtc gacaaactgt
     1861 tcatccagtt agtacaaacc tataatcagt tgtttgaaga gaaccctata aatgcaagtg
     1921 gcgtggatgc gaaggctatt cttagcgccc gcctctctaa atcccgacgg ctagaaaacc
     1981 tgatcgcaca attacccgga gagaagaaaa atgggttgtt cggtaacctt atagcgctct
     2041 cactaggcct gacaccaaat tttaagtcga acttcgactt agctgaagat gccaaattgc
     2101 agcttagtaa ggacacgtac gatgacgatc tcgacaatct actggcacaa attggagatc
     2161 agtatgcgga cttatttttg gctgccaaaa accttagcga tgcaatcctc ctatctgaca
     2221 tactgagagt taatactgag attaccaagg cgccgttatc cgcttcaatg atcaaaaggt
     2281 acgatgaaca tcaccaagac ttgacacttc tcaaggccct agtccgtcag caactgcctg
     2341 agaaatataa ggaaatattc tttgatcagt cgaaaaacgg gtacgcaggt tatattgacg
     2401 gcggagcgag tcaagaggaa ttctacaagt ttatcaaacc catattagag aagatggatg
     2461 ggacggaaga gttgcttgta aaactcaatc gcgaagatct actgcgaaag cagcggactt
     2521 tcgacaacgg tagcattcca catcaaatcc acttaggcga attgcatgct atacttagaa
     2581 ggcaggagga tttttatccg ttcctcaaag acaatcgtga aaagattgag aaaatcctaa
     2641 cctttcgcat accttactat gtgggacccc tggcccgagg gaactctcgg ttcgcatgga
     2701 tgacaagaaa gtccgaagaa acgattactc catggaattt tgaggaagtt gtcgataaag
     2761 gtgcgtcagc tcaatcgttc atcgagagga tgaccaactt tgacaagaat ttaccgaacg
     2821 aaaaagtatt gcctaagcac agtttacttt acgagtattt cacagtgtac aatgaactca
     2881 cgaaagttaa gtatgtcact gagggcatgc gtaaacccgc ctttctaagc ggagaacaga
     2941 agaaagcaat agtagatctg ttattcaaga ccaaccgcaa agtgacagtt aagcaattga
     3001 aagaggacta ctttaagaaa attgaatgct tcgattctgt cgagatctcc ggggtagaag
     3061 atcgatttaa tgcgtcactt ggtacgtatc atgacctcct aaagataatt aaagataagg
     3121 acttcctgga taacgaagag aatgaagata tcttagaaga tatagtgttg actcttaccc
     3181 tctttgaaga tcgggaaatg attgaggaaa gactaaaaac atacgctcac ctgttcgacg
     3241 ataaggttat gaaacagtta aagaggcgtc gctatacggg ctggggacga ttgtcgcgga
     3301 aacttatcaa cgggataaga gacaagcaaa gtggtaaaac tattctcgat tttctaaaga
     3361 gcgacggctt cgccaatagg aactttatgc agctgatcca tgatgactct ttaaccttca
     3421 aagaggatat acaaaaggca caggtttccg gacaagggga ctcattgcac gaacatattg
     3481 cgaatcttgc tggttcgcca gccatcaaaa agggcatact ccagacagtc aaagtagtgg
     3541 atgagctagt taaggtcatg ggacgtcaca aaccggaaaa cattgtaatc gagatggcac
     3601 gcgaaaatca aacgactcag aaggggcaaa aaaacagtcg agagcggatg aagagaatag
     3661 aagagggtat taaagaactg ggcagccaga tcttaaagga gcatcctgtg gaaaataccc
     3721 aattgcagaa cgagaaactt tacctctatt acctacaaaa tggaagggac atgtatgttg
     3781 atcaggaact ggacataaac cgtttatctg attacgacgt cgatgccatt gtaccccaat
     3841 cctttttgaa ggacgattca atcgacaata aagtgcttac acgctcggat aagaaccgag
     3901 ggaaaagtga caatgttcca agcgaggaag tcgtaaagaa aatgaagaac tattggcggc
     3961 agctcctaaa tgcgaaactg ataacgcaaa gaaagttcga taacttaact aaagctgaga
     4021 ggggtggctt gtctgaactt gacaaggccg gatttattaa acgtcagctc gtggaaaccc
     4081 gccaaatcac aaagcatgtt gcacagatac tagattcccg aatgaatacg aaatacgacg
     4141 agaacgataa gctgattcgg gaagtcaaag taatcacttt aaagtcaaaa ttggtgtcgg
     4201 acttcagaaa ggattttcaa ttctataaag ttagggagat aaataactac caccatgcgc
     4261 acgacgctta tcttaatgcc gtcgtaggga ccgcactcat taagaaatac ccgaagctag
     4321 aaagtgagtt tgtgtatggt gattacaaag tttatgacgt ccgtaagatg atcgcgaaaa
     4381 gcgaacagga gataggcaag gctacagcca aatacttctt ttattctaac attatgaatt
     4441 tctttaagac ggaaatcact ctggcaaacg gagagatacg caaacgacct ttaattgaaa
     4501 ccaatgggga gacaggtgaa atcgtatggg ataagggccg ggacttcgcg acggtgagaa
     4561 aagttttgtc catgccccaa gtcaacatag taaagaaaac tgaggtgcag accggagggt
     4621 tttcaaagga atcgattctt ccaaaaagga atagtgataa gctcatcgct cgtaaaaagg
     4681 actgggaccc gaaaaagtac ggtggcttcg atagccctac agttgcctat tctgtcctag
     4741 tagtggcaaa agttgagaag ggaaaatcca agaaactgaa gtcagtcaaa gaattattgg
     4801 ggataacgat tatggagcgc tcgtcttttg aaaagaaccc catcgacttc cttgaggcga
     4861 aaggttacaa ggaagtaaaa aaggatctca taattaaact accaaagtat agtctgtttg
     4921 agttagaaaa tggccgaaaa cggatgttgg ctagcgccgg agagcttcaa aaggggaacg
     4981 aactcgcact accgtctaaa tacgtgaatt tcctgtattt agcgtcccat tacgagaagt
     5041 tgaaaggttc acctgaagat aacgaacaga agcaactttt tgttgagcag cacaaacatt
     5101 atctcgacga aatcatagag caaatttcgg aattcagtaa gagagtcatc ctagctgatg
     5161 ccaatctgga caaagtatta agcgcataca acaagcacag ggataaaccc atacgtgagc
     5221 aggcggaaaa tattatccat ttgtttactc ttaccaacct cggcgctcca gccgcattca
     5281 agtattttga cacaacgata gatcgcaaac gatacacttc taccaaggag gtgctagacg
     5341 cgacactgat tcaccaatcc atcacgggat tatatgaaac tcggatagat ttgtcacagc
     5401 ttgggggtga cggatccccc aagaagaaga ggaaagtctc gagcgactac aaagaccatg
     5461 acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag gctgcaggag
     5521 gcggaggtag cccatcgggt gtggagggcg cagctttcca gagccgactt cctcatgacc
     5581 ggatgacttc tcaagaagca gcctgttttc cagatattat cagtggacca caacagaccc
     5641 agaaggtttt tcttttcatt agaaaccgca cactgcagtt gtggttggat aatccaaaga
     5701 ttcagctgac atttgaggct actctccaac aattagaagc accttataac agtgatactg
     5761 tgcttgtcca ccgagttcac agttatttag agcgtcatgg tcttatcaac ttcggcatct
     5821 ataagaggat aaaaccccta ccaactaaaa agacaggaaa ggtaattatt ataggctctg
     5881 gggtctcagg cttggcagca gctcgacagt tacaaagttt tggaatggat gtcacacttt
     5941 tggaagccag ggatcgtgtg ggtggacgag ttgccacatt tcgcaaagga aactatgtag
     6001 ctgatcttgg agccatggtg gtaacaggtc ttggagggaa tcctatggct gtggtcagca
     6061 aacaagtaaa tatggaactg gccaagatca agcaaaaatg cccactttat gaagccaacg
     6121 gacaagctgt tcctaaagag aaagatgaaa tggtagagca agagtttaac cggttgctag
     6181 aagctacatc ttaccttagt catcaactag acttcaatgt cctcaataat aagcctgtgt
     6241 cccttggcca ggcattggaa gttgtcattc agttacaaga gaagcatgtc aaagatgagc
     6301 agattgaaca ttggaagaag atagtgaaaa ctcaggaaga attgaaagaa cttcttaata
     6361 agatggtaaa tttgaaagag aaaattaaag aactccatca gcaatacaaa gaagcatctg
     6421 aagtaaagcc acccagagat attactgccg agttcttagt gaaaagcaaa cacagggatc
     6481 tgaccgccct atgcaaggaa tatgatgaat tagctgaaac acaaggaaag ctagaagaaa
     6541 aacttcagga gttggaagcg aatcccccaa gtgatgtata tctctcatca agagacagac
     6601 aaatacttga ttggcatttt gcaaatcttg aatttgctaa tgccacacct ctctcaactc
     6661 tctcccttaa gcactgggat caggatgatg actttgagtt cactggcagc cacctgacag
     6721 taaggaatgg ctactcgtgt gtgcctgtgg ctttagcaga aggcctagac attaaactga
     6781 atacagcagt gcgacaggtt cgctacacgg cttcaggatg tgaagtgata gctgtgaata
     6841 cccgctccac gagtcaaacc tttatttata aatgcgacgc agttctctgt acccttcccc
     6901 tgggtgtgct gaagcagcag ccaccagccg ttcagtttgt gccacctctc cctgagtgga
     6961 aaacatctgc agtccaaagg atgggatttg gcaaccttaa cgcagtggtg ttgtgttttg
     7021 atcgggtgtt ctgggatcca agtgtcaatt tgttcgggca tgttggcagt acgactgcca
     7081 gcaggggtga gctcttcctc ttctggaacc tctataaagc tccaatactg ttggcactag
     7141 tggcaggaga agctgctggt atcatggaaa acataagtga cgatgtgatt gttggccgat
     7201 gcctggccat tctcaaaggg atttttggta gcagtgcagt acctcagccc aaagaaactg
     7261 tggtgtctcg ttggcgtgct gatccctggg ctcggggctc ttattcctat gttgctgcag
     7321 gatcatctgg aaatgactat gatttaatgg ctcagccaat cactcctggc ccctcgattc
     7381 caggtgcccc acagccgatt ccacgactct tctttgcggg agaacatacg atccgtaact
     7441 acccagccac agtgcatggt gctctgctga gtgggctgcg agaagcggga agaattgcag
     7501 accagttttt gggggccatg tatacgctgc ctcgccaggc cacaccaggt gttcctgcac
     7561 agcagtcccc aagcatgtga aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     7621 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     7681 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     7741 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     7801 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     7861 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     7981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     8041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     8101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8701 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     9001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     9061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     9121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     9181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     9241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
    10021 gcgcacattt ccccgaaaag tgccacctga cgtcgacggg tatacagaca tgataagata
    10081 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
    10141 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt
    10201 gggcgaagaa ctctcggcat ctactctatt cctttgccct cggacgagtg ctggggcgtc
    10261 ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc
    10321 gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac
    10381 cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa
    10441 gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc
    10501 gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt
    10561 tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta
    10621 tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga
    10681 cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac
    10741 tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata
    10801 tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc
    10861 tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc ggctgcagaa
    10921 cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga
    10981 tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga atcgggagcg
    11041 cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat
    11101 ttacccgcag gacatatcca cg
//
