LOCUS       sequence-204224-        9967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-dDNMT3a fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9967)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9967)
  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            "Unpublished"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-dDNMT3a-C706S-Hygro
FEATURES             Location/Qualifiers
     source          1..9967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     315..338
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        378..757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        758..961
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     912..932
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1003..1022
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1003..1021
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1029..1038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1035..4436,4437..4439,4440..5036,5037,5038..5040,
                     5041..5042,5043..5045,5046..5138)
                     /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(1035..1061,1062..1064,1065..3551,3552..3554,
                     3555..5138)
                     /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             5145..5165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5172..5237
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6168..6185)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6174..6398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6469..6485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6469..6485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6482..6504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6493..6509
                     /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(6517..6523,6524..6541,6542..6547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6562..6583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6700..6717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6871..7459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6951..6970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7630..8421,8422..8490))
                     /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     8253..8272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8491..8595)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8646..8767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8657..8676)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8711..8730
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8798..9823)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8798..9823)
                     /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(9841..9967,1..373))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9889..9908)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 cccccggacc caccccttcc cagcctctga gcccagaaag cgaaggagca aagctgctat
       61 tggccgctgc cccaaaggcc tacccgcttc cattgctcag cggtgctgtc catctgcacg
      121 agactagtga gacgtgctac ttccatttgt cacgtcctgc acgacgcgag ctgcggggcg
      181 ggggggaact tcctgactag gggaggagta gaaggtggcg cgaaggggcc accaaagaac
      241 ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc gaggccagag gccacttgtg
      301 tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc
      361 gcctccccta cccggttgac attgattatt gactagttat taatagtaat caattacggg
      421 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      481 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      541 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      601 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      661 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      721 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      781 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      841 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      901 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      961 tggtttagtg aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag
     1021 ggagagccgc caccatggat aaaaagtatt ctattggttt agccatcggc actaattccg
     1081 ttggatgggc tgtcataacc gatgaataca aagtaccttc aaagaaattt aaggtgttgg
     1141 ggaacacaga ccgtcattcg attaaaaaga atcttatcgg tgccctccta ttcgatagtg
     1201 gcgaaacggc agaggcgact cgcctgaaac gaaccgctcg gagaaggtat acacgtcgca
     1261 agaaccgaat atgttactta caagaaattt ttagcaatga gatggccaaa gttgacgatt
     1321 ctttctttca ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa catgaacggc
     1381 accccatctt tggaaacata gtagatgagg tggcatatca tgaaaagtac ccaacgattt
     1441 atcacctcag aaaaaagcta gttgactcaa ctgataaagc ggacctgagg ttaatctact
     1501 tggctcttgc ccatatgata aagttccgtg ggcactttct cattgagggt gatctaaatc
     1561 cggacaactc ggatgtcgac aaactgttca tccagttagt acaaacctat aatcagttgt
     1621 ttgaagagaa ccctataaat gcaagtggcg tggatgcgaa ggctattctt agcgcccgcc
     1681 tctctaaatc ccgacggcta gaaaacctga tcgcacaatt acccggagag aagaaaaatg
     1741 ggttgttcgg taaccttata gcgctctcac taggcctgac accaaatttt aagtcgaact
     1801 tcgacttagc tgaagatgcc aaattgcagc ttagtaagga cacgtacgat gacgatctcg
     1861 acaatctact ggcacaaatt ggagatcagt atgcggactt atttttggct gccaaaaacc
     1921 ttagcgatgc aatcctccta tctgacatac tgagagttaa tactgagatt accaaggcgc
     1981 cgttatccgc ttcaatgatc aaaaggtacg atgaacatca ccaagacttg acacttctca
     2041 aggccctagt ccgtcagcaa ctgcctgaga aatataagga aatattcttt gatcagtcga
     2101 aaaacgggta cgcaggttat attgacggcg gagcgagtca agaggaattc tacaagttta
     2161 tcaaacccat attagagaag atggatggga cggaagagtt gcttgtaaaa ctcaatcgcg
     2221 aagatctact gcgaaagcag cggactttcg acaacggtag cattccacat caaatccact
     2281 taggcgaatt gcatgctata cttagaaggc aggaggattt ttatccgttc ctcaaagaca
     2341 atcgtgaaaa gattgagaaa atcctaacct ttcgcatacc ttactatgtg ggacccctgg
     2401 cccgagggaa ctctcggttc gcatggatga caagaaagtc cgaagaaacg attactccat
     2461 ggaattttga ggaagttgtc gataaaggtg cgtcagctca atcgttcatc gagaggatga
     2521 ccaactttga caagaattta ccgaacgaaa aagtattgcc taagcacagt ttactttacg
     2581 agtatttcac agtgtacaat gaactcacga aagttaagta tgtcactgag ggcatgcgta
     2641 aacccgcctt tctaagcgga gaacagaaga aagcaatagt agatctgtta ttcaagacca
     2701 accgcaaagt gacagttaag caattgaaag aggactactt taagaaaatt gaatgcttcg
     2761 attctgtcga gatctccggg gtagaagatc gatttaatgc gtcacttggt acgtatcatg
     2821 acctcctaaa gataattaaa gataaggact tcctggataa cgaagagaat gaagatatct
     2881 tagaagatat agtgttgact cttaccctct ttgaagatcg ggaaatgatt gaggaaagac
     2941 taaaaacata cgctcacctg ttcgacgata aggttatgaa acagttaaag aggcgtcgct
     3001 atacgggctg gggacgattg tcgcggaaac ttatcaacgg gataagagac aagcaaagtg
     3061 gtaaaactat tctcgatttt ctaaagagcg acggcttcgc caataggaac tttatgcagc
     3121 tgatccatga tgactcttta accttcaaag aggatataca aaaggcacag gtttccggac
     3181 aaggggactc attgcacgaa catattgcga atcttgctgg ttcgccagcc atcaaaaagg
     3241 gcatactcca gacagtcaaa gtagtggatg agctagttaa ggtcatggga cgtcacaaac
     3301 cggaaaacat tgtaatcgag atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa
     3361 acagtcgaga gcggatgaag agaatagaag agggtattaa agaactgggc agccagatct
     3421 taaaggagca tcctgtggaa aatacccaat tgcagaacga gaaactttac ctctattacc
     3481 tacaaaatgg aagggacatg tatgttgatc aggaactgga cataaaccgt ttatctgatt
     3541 acgacgtcga tgccattgta ccccaatcct ttttgaagga cgattcaatc gacaataaag
     3601 tgcttacacg ctcggataag aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg
     3661 taaagaaaat gaagaactat tggcggcagc tcctaaatgc gaaactgata acgcaaagaa
     3721 agttcgataa cttaactaaa gctgagaggg gtggcttgtc tgaacttgac aaggccggat
     3781 ttattaaacg tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca cagatactag
     3841 attcccgaat gaatacgaaa tacgacgaga acgataagct gattcgggaa gtcaaagtaa
     3901 tcactttaaa gtcaaaattg gtgtcggact tcagaaagga ttttcaattc tataaagtta
     3961 gggagataaa taactaccac catgcgcacg acgcttatct taatgccgtc gtagggaccg
     4021 cactcattaa gaaatacccg aagctagaaa gtgagtttgt gtatggtgat tacaaagttt
     4081 atgacgtccg taagatgatc gcgaaaagcg aacaggagat aggcaaggct acagccaaat
     4141 acttctttta ttctaacatt atgaatttct ttaagacgga aatcactctg gcaaacggag
     4201 agatacgcaa acgaccttta attgaaacca atggggagac aggtgaaatc gtatgggata
     4261 agggccggga cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc aacatagtaa
     4321 agaaaactga ggtgcagacc ggagggtttt caaaggaatc gattcttcca aaaaggaata
     4381 gtgataagct catcgctcgt aaaaaggact gggacccgaa aaagtacggt ggcttcgata
     4441 gccctacagt tgcctattct gtcctagtag tggcaaaagt tgagaaggga aaatccaaga
     4501 aactgaagtc agtcaaagaa ttattgggga taacgattat ggagcgctcg tcttttgaaa
     4561 agaaccccat cgacttcctt gaggcgaaag gttacaagga agtaaaaaag gatctcataa
     4621 ttaaactacc aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta
     4681 gcgccggaga gcttcaaaag gggaacgaac tcgcactacc gtctaaatac gtgaatttcc
     4741 tgtatttagc gtcccattac gagaagttga aaggttcacc tgaagataac gaacagaagc
     4801 aactttttgt tgagcagcac aaacattatc tcgacgaaat catagagcaa atttcggaat
     4861 tcagtaagag agtcatccta gctgatgcca atctggacaa agtattaagc gcatacaaca
     4921 agcacaggga taaacccata cgtgagcagg cggaaaatat tatccatttg tttactctta
     4981 ccaacctcgg cgctccagcc gcattcaagt attttgacac aacgatagat cgcaaacgat
     5041 acacttctac caaggaggtg ctagacgcga cactgattca ccaatccatc acgggattat
     5101 atgaaactcg gatagatttg tcacagcttg ggggtgacgg atcccccaag aagaagagga
     5161 aagtctcgag cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca
     5221 aggatgacga tgacaaggct gcaggaggcg gaggtagcaa ccacgaccag gaatttgacc
     5281 ctccaaaggt ttacccacct gtcccagctg agaagaggaa gcccatccgg gtgctgtctc
     5341 tctttgatgg aatcgctaca gggctcctgg tgctgaagga cttgggcatt caggtggacc
     5401 gctacattgc ctcggaggtg tgtgaggact ccatcacggt gggcatggtg cggcaccagg
     5461 ggaagatcat gtacgtcggg gacgtccgca gcgtcacaca gaagcatatc caggagtggg
     5521 gcccattcga tctggtgatt gggggcagtc cttcgaatga cctctccatc gtcaaccctg
     5581 ctcgcaaggg cctctacgag ggcactggcc ggctcttctt tgagttctac cgcctcctgc
     5641 atgatgcgcg gcccaaggag ggagatgatc gccccttctt ctggctcttt gagaatgtgg
     5701 tggccatggg cgttagtgac aagagggaca tctcgcgatt tctcgagtcc aaccctgtga
     5761 tgattgatgc caaagaagtg tcagctgcac acagggcccg ctacttctgg ggtaaccttc
     5821 ccggtatgaa caggccgttg gcatccactg tgaatgataa gctggagctg caggagtgtc
     5881 tggagcatgg caggatagcc aagttcagca aagtgaggac cattactacg aggtcaaact
     5941 ccataaagca gggcaaagac cagcattttc ctgtcttcat gaatgagaaa gaggacatct
     6001 tatggtgcac tgaaatggaa agggtatttg gtttcccagt ccactatact gacgtctcca
     6061 acatgagccg cttggcgagg cagagactgc tgggccggtc atggagcgtg ccagtcatcc
     6121 gccacctctt cgctccgctg aaggagtatt gaaaacccgc tgatcagcct cgactgtgcc
     6181 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6241 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     6301 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     6361 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     6421 ctggggctcg ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6481 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6541 gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg ggtatacaga
     8641 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     8701 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     8761 acaagttggg gtgggcgaag aactctcggc atctactcta ttcctttgcc ctcggacgag
     8821 tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg gtccagacgg
     8881 ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat cggacgattg
     8941 cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc aagctctgat
     9001 agagttggtc aagaccaatg cggagcatat acgcccggag ccgcggcgat cctgcaagct
     9061 ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac cacggcctcc
     9121 agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg ctccagtcaa
     9181 tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca
     9241 cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg agagcctgcg
     9301 cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca tggggatcag
     9361 caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc ggtccgaatg
     9421 ggccgaaccc gctcgtctgg ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga
     9481 ccggctgcag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg acaccctgtg
     9541 cacggcggga gatgcaatag gtcaggctct cgctgaattc cccaatgtca agcacttccg
     9601 gaatcgggag cgcggccgat gcaaagtgcc gataaacata acgatctttg tagaaaccat
     9661 cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac
     9721 gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac ttttcgatca
     9781 gaaacttctc gacagacgtc gcggtgagtt caggcttttt catggtggcg ggatgcaggt
     9841 cgaaaggccc ggagatgagg aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt
     9901 gcagaatgcc gggcctccgg aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct
     9961 gagcccg
//
