LOCUS       sequence-204471-       10219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-HDAC1 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  JOURNAL   Unpublished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            "Unpublished"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-Hygro
FEATURES             Location/Qualifiers
     source          1..10219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..47)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     421..444
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        484..863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        864..1067
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1018..1038
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1109..1128
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1109..1127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1135..1144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1141..4542,4543..4545,4546..5142,5143,5144..5146,
                     5147..5148,5149..5151,5152..5244)
                     /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(1141..1167,1168..1170,1171..3657,3658..3660,
                     3661..5244)
                     /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             5251..5271
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5278..5343
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6526..6543)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6532..6756
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6827..6843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6827..6843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6840..6862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6851..6867
                     /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(6875..6881,6882..6899,6900..6905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6920..6941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7058..7075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7229..7817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7309..7328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7988..8779,8780..8848))
                     /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     8611..8630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8849..8953)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9004..9125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9015..9034)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9069..9088
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9156..10181)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(9156..10181)
                     /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(10199..10219,1..479))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga
       61 ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc cggacccacc
      121 ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc cgctgcccca
      181 aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac tagtgagacg
      241 tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg ggaacttcct
      301 gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag ccggttggcg
      361 cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc gccaagtgcc
      421 cagcggggct gctaaagcgc atgctccaga ctgccttggg aaaagcgcct cccctacccg
      481 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      541 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      601 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      661 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      721 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      781 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      841 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      901 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      961 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     1021 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     1081 gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag agccgccacc
     1141 atggataaaa agtattctat tggtttagcc atcggcacta attccgttgg atgggctgtc
     1201 ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa cacagaccgt
     1261 cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga aacggcagag
     1321 gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa ccgaatatgt
     1381 tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt ctttcaccgt
     1441 ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc catctttgga
     1501 aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca cctcagaaaa
     1561 aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc tcttgcccat
     1621 atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga caactcggat
     1681 gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga agagaaccct
     1741 ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc taaatcccga
     1801 cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt gttcggtaac
     1861 cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga cttagctgaa
     1921 gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa tctactggca
     1981 caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag cgatgcaatc
     2041 ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt atccgcttca
     2101 atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc cctagtccgt
     2161 cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa cgggtacgca
     2221 ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa acccatatta
     2281 gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga tctactgcga
     2341 aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg cgaattgcat
     2401 gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg tgaaaagatt
     2461 gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg agggaactct
     2521 cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccatggaa ttttgaggaa
     2581 gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa ctttgacaag
     2641 aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg
     2701 tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta
     2761 agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca
     2821 gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc tgtcgagatc
     2881 tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct cctaaagata
     2941 attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga agatatagtg
     3001 ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa aacatacgct
     3061 cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac gggctgggga
     3121 cgattgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa aactattctc
     3181 gattttctaa agagcgacgg cttcgccaat aggaacttta tgcagctgat ccatgatgac
     3241 tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg ggactcattg
     3301 cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat actccagaca
     3361 gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga aaacattgta
     3421 atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag tcgagagcgg
     3481 atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa ggagcatcct
     3541 gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca aaatggaagg
     3601 gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga cgtcgatgcc
     3661 attgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct tacacgctcg
     3721 gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa gaaaatgaag
     3781 aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt cgataactta
     3841 actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat taaacgtcag
     3901 ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga tactagattc ccgaatgaat
     3961 acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac tttaaagtca
     4021 aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga gataaataac
     4081 taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact cattaagaaa
     4141 tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga cgtccgtaag
     4201 atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt cttttattct
     4261 aacattatga atttctttaa gacggaaatc actctggcaa acggagagat acgcaaacga
     4321 cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg ccgggacttc
     4381 gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa aactgaggtg
     4441 cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga taagctcatc
     4501 gctcgtaaaa aggactggga cccgaaaaag tacggtggct tcgatagccc tacagttgcc
     4561 tattctgtcc tagtagtggc aaaagttgag aagggaaaat ccaagaaact gaagtcagtc
     4621 aaagaattat tggggataac gattatggag cgctcgtctt ttgaaaagaa ccccatcgac
     4681 ttccttgagg cgaaaggtta caaggaagta aaaaaggatc tcataattaa actaccaaag
     4741 tatagtctgt ttgagttaga aaatggccga aaacggatgt tggctagcgc cggagagctt
     4801 caaaagggga acgaactcgc actaccgtct aaatacgtga atttcctgta tttagcgtcc
     4861 cattacgaga agttgaaagg ttcacctgaa gataacgaac agaagcaact ttttgttgag
     4921 cagcacaaac attatctcga cgaaatcata gagcaaattt cggaattcag taagagagtc
     4981 atcctagctg atgccaatct ggacaaagta ttaagcgcat acaacaagca cagggataaa
     5041 cccatacgtg agcaggcgga aaatattatc catttgttta ctcttaccaa cctcggcgct
     5101 ccagccgcat tcaagtattt tgacacaacg atagatcgca aacgatacac ttctaccaag
     5161 gaggtgctag acgcgacact gattcaccaa tccatcacgg gattatatga aactcggata
     5221 gatttgtcac agcttggggg tgacggatcc cccaagaaga agaggaaagt ctcgagcgac
     5281 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     5341 aaggctgcag gaggcggagg tagcacgcag ggcacccgga ggaaagtctg ttactactac
     5401 gacggggatg ttggaaatta ctattatgga caaggccacc caatgaagcc tcaccgaatc
     5461 cgcatgactc ataatttgct gctcaactat ggtctctacc gaaaaatgga aatctatcgc
     5521 cctcacaaag ccaatgctga ggagatgacc aagtaccaca gcgatgacta cattaaattc
     5581 ttgcgctcca tccgtccaga taacatgtcg gagtacagca agcagatgca gagattcaac
     5641 gttggtgagg actgtccagt attcgatggc ctgtttgagt tctgtcagtt gtctactggt
     5701 ggttctgtgg caagtgctgt gaaacttaat aagcagcaga cggacatcgc tgtgaattgg
     5761 gctgggggcc tgcaccatgc aaagaagtcc gaggcatctg gcttctgtta cgtcaatgat
     5821 atcgtcttgg ccatcctgga actgctaaag tatcaccaga gggtgctgta cattgacatt
     5881 gatattcacc atggtgacgg cgtggaagag gccttctaca ccacggaccg ggtcatgact
     5941 gtgtcctttc ataagtatgg agagtacttc ccaggaactg gggacctacg ggatatcggg
     6001 gctggcaaag gcaagtatta tgctgttaac tacccgctcc gagacgggat tgatgacgag
     6061 tcctatgagg ccattttcaa gccggtcatg tccaaagtaa tggagatgtt ccagcctagt
     6121 gcggtggtct tacagtgtgg ctcagactcc ctatctgggg atcggttagg ttgcttcaat
     6181 ctaactatca aaggacacgc caagtgtgtg gaatttgtca agagctttaa cctgcctatg
     6241 ctgatgctgg gaggcggtgg ttacaccatt cgtaacgttg cccggtgctg gacatatgag
     6301 acagctgtgg ccctggatac ggagatccct aatgagcttc catacaatga ctactttgaa
     6361 tactttggac cagatttcaa gctccacatc agtccttcca atatgactaa ccagaacacg
     6421 aatgagtacc tggagaagat caaacagcga ctgtttgaga accttagaat gctgccgcac
     6481 gcacctgggg tccaaatgca ggcgatttga aaacccgctg atcagcctcg actgtgcctt
     6541 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6601 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6661 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6721 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     6781 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6841 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6901 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7021 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7081 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7141 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacggg tatacagaca
     9001 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     9061 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     9121 aagttggggt gggcgaagaa ctctcggcat ctactctatt cctttgccct cggacgagtg
     9181 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc
     9241 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg
     9301 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag
     9361 agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc
     9421 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag
     9481 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg
     9541 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg
     9601 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg
     9661 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca
     9721 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg
     9781 ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc
     9841 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca
     9901 cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga
     9961 atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg
    10021 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga
    10081 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga
    10141 aacttctcga cagacgtcgc ggtgagttca ggctttttca tggtggcggg atgcaggtcg
    10201 aaaggcccgg agatgagga
//
