LOCUS       sequence-204454-       11233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-TET1 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11233)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11233)
  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            "Unpublished"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-TET1-Hygro
FEATURES             Location/Qualifiers
     source          1..11233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     294..317
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        357..736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        737..940
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     891..911
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     982..1001
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        982..1000
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1008..1017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1014..4415,4416..4418,4419..5015,5016,5017..5019,
                     5020..5021,5022..5024,5025..5117)
                     /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(1014..1040,1041..1043,1044..3530,3531..3533,
                     3534..5117)
                     /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             5124..5144
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5151..5216
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7413..7430)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7419..7643
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7714..7730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7714..7730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7727..7749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7738..7754
                     /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(7762..7768,7769..7786,7787..7792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7807..7828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7945..7962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8116..8704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8196..8215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8875..9666,9667..9735))
                     /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     9498..9517
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9736..9840)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9891..10012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9902..9921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9956..9975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(10043..11068)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(10043..11068)
                     /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(11086..11233,1..352))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(11134..11153)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct
       61 acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag acgtgctact
      121 tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt cctgactagg
      181 ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg gcgcctaccg
      241 gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt gcccagcggg
      301 gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac ccggttgaca
      361 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      421 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      481 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      541 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      601 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      661 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      721 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      781 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      841 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      901 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      961 ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc accatggata
     1021 aaaagtattc tattggttta gccatcggca ctaattccgt tggatgggct gtcataaccg
     1081 atgaatacaa agtaccttca aagaaattta aggtgttggg gaacacagac cgtcattcga
     1141 ttaaaaagaa tcttatcggt gccctcctat tcgatagtgg cgaaacggca gaggcgactc
     1201 gcctgaaacg aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata tgttacttac
     1261 aagaaatttt tagcaatgag atggccaaag ttgacgattc tttctttcac cgtttggaag
     1321 agtccttcct tgtcgaagag gacaagaaac atgaacggca ccccatcttt ggaaacatag
     1381 tagatgaggt ggcatatcat gaaaagtacc caacgattta tcacctcaga aaaaagctag
     1441 ttgactcaac tgataaagcg gacctgaggt taatctactt ggctcttgcc catatgataa
     1501 agttccgtgg gcactttctc attgagggtg atctaaatcc ggacaactcg gatgtcgaca
     1561 aactgttcat ccagttagta caaacctata atcagttgtt tgaagagaac cctataaatg
     1621 caagtggcgt ggatgcgaag gctattctta gcgcccgcct ctctaaatcc cgacggctag
     1681 aaaacctgat cgcacaatta cccggagaga agaaaaatgg gttgttcggt aaccttatag
     1741 cgctctcact aggcctgaca ccaaatttta agtcgaactt cgacttagct gaagatgcca
     1801 aattgcagct tagtaaggac acgtacgatg acgatctcga caatctactg gcacaaattg
     1861 gagatcagta tgcggactta tttttggctg ccaaaaacct tagcgatgca atcctcctat
     1921 ctgacatact gagagttaat actgagatta ccaaggcgcc gttatccgct tcaatgatca
     1981 aaaggtacga tgaacatcac caagacttga cacttctcaa ggccctagtc cgtcagcaac
     2041 tgcctgagaa atataaggaa atattctttg atcagtcgaa aaacgggtac gcaggttata
     2101 ttgacggcgg agcgagtcaa gaggaattct acaagtttat caaacccata ttagagaaga
     2161 tggatgggac ggaagagttg cttgtaaaac tcaatcgcga agatctactg cgaaagcagc
     2221 ggactttcga caacggtagc attccacatc aaatccactt aggcgaattg catgctatac
     2281 ttagaaggca ggaggatttt tatccgttcc tcaaagacaa tcgtgaaaag attgagaaaa
     2341 tcctaacctt tcgcatacct tactatgtgg gacccctggc ccgagggaac tctcggttcg
     2401 catggatgac aagaaagtcc gaagaaacga ttactccatg gaattttgag gaagttgtcg
     2461 ataaaggtgc gtcagctcaa tcgttcatcg agaggatgac caactttgac aagaatttac
     2521 cgaacgaaaa agtattgcct aagcacagtt tactttacga gtatttcaca gtgtacaatg
     2581 aactcacgaa agttaagtat gtcactgagg gcatgcgtaa acccgccttt ctaagcggag
     2641 aacagaagaa agcaatagta gatctgttat tcaagaccaa ccgcaaagtg acagttaagc
     2701 aattgaaaga ggactacttt aagaaaattg aatgcttcga ttctgtcgag atctccgggg
     2761 tagaagatcg atttaatgcg tcacttggta cgtatcatga cctcctaaag ataattaaag
     2821 ataaggactt cctggataac gaagagaatg aagatatctt agaagatata gtgttgactc
     2881 ttaccctctt tgaagatcgg gaaatgattg aggaaagact aaaaacatac gctcacctgt
     2941 tcgacgataa ggttatgaaa cagttaaaga ggcgtcgcta tacgggctgg ggacgattgt
     3001 cgcggaaact tatcaacggg ataagagaca agcaaagtgg taaaactatt ctcgattttc
     3061 taaagagcga cggcttcgcc aataggaact ttatgcagct gatccatgat gactctttaa
     3121 ccttcaaaga ggatatacaa aaggcacagg tttccggaca aggggactca ttgcacgaac
     3181 atattgcgaa tcttgctggt tcgccagcca tcaaaaaggg catactccag acagtcaaag
     3241 tagtggatga gctagttaag gtcatgggac gtcacaaacc ggaaaacatt gtaatcgaga
     3301 tggcacgcga aaatcaaacg actcagaagg ggcaaaaaaa cagtcgagag cggatgaaga
     3361 gaatagaaga gggtattaaa gaactgggca gccagatctt aaaggagcat cctgtggaaa
     3421 atacccaatt gcagaacgag aaactttacc tctattacct acaaaatgga agggacatgt
     3481 atgttgatca ggaactggac ataaaccgtt tatctgatta cgacgtcgat gccattgtac
     3541 cccaatcctt tttgaaggac gattcaatcg acaataaagt gcttacacgc tcggataaga
     3601 accgagggaa aagtgacaat gttccaagcg aggaagtcgt aaagaaaatg aagaactatt
     3661 ggcggcagct cctaaatgcg aaactgataa cgcaaagaaa gttcgataac ttaactaaag
     3721 ctgagagggg tggcttgtct gaacttgaca aggccggatt tattaaacgt cagctcgtgg
     3781 aaacccgcca aatcacaaag catgttgcac agatactaga ttcccgaatg aatacgaaat
     3841 acgacgagaa cgataagctg attcgggaag tcaaagtaat cactttaaag tcaaaattgg
     3901 tgtcggactt cagaaaggat tttcaattct ataaagttag ggagataaat aactaccacc
     3961 atgcgcacga cgcttatctt aatgccgtcg tagggaccgc actcattaag aaatacccga
     4021 agctagaaag tgagtttgtg tatggtgatt acaaagttta tgacgtccgt aagatgatcg
     4081 cgaaaagcga acaggagata ggcaaggcta cagccaaata cttcttttat tctaacatta
     4141 tgaatttctt taagacggaa atcactctgg caaacggaga gatacgcaaa cgacctttaa
     4201 ttgaaaccaa tggggagaca ggtgaaatcg tatgggataa gggccgggac ttcgcgacgg
     4261 tgagaaaagt tttgtccatg ccccaagtca acatagtaaa gaaaactgag gtgcagaccg
     4321 gagggttttc aaaggaatcg attcttccaa aaaggaatag tgataagctc atcgctcgta
     4381 aaaaggactg ggacccgaaa aagtacggtg gcttcgatag ccctacagtt gcctattctg
     4441 tcctagtagt ggcaaaagtt gagaagggaa aatccaagaa actgaagtca gtcaaagaat
     4501 tattggggat aacgattatg gagcgctcgt cttttgaaaa gaaccccatc gacttccttg
     4561 aggcgaaagg ttacaaggaa gtaaaaaagg atctcataat taaactacca aagtatagtc
     4621 tgtttgagtt agaaaatggc cgaaaacgga tgttggctag cgccggagag cttcaaaagg
     4681 ggaacgaact cgcactaccg tctaaatacg tgaatttcct gtatttagcg tcccattacg
     4741 agaagttgaa aggttcacct gaagataacg aacagaagca actttttgtt gagcagcaca
     4801 aacattatct cgacgaaatc atagagcaaa tttcggaatt cagtaagaga gtcatcctag
     4861 ctgatgccaa tctggacaaa gtattaagcg catacaacaa gcacagggat aaacccatac
     4921 gtgagcaggc ggaaaatatt atccatttgt ttactcttac caacctcggc gctccagccg
     4981 cattcaagta ttttgacaca acgatagatc gcaaacgata cacttctacc aaggaggtgc
     5041 tagacgcgac actgattcac caatccatca cgggattata tgaaactcgg atagatttgt
     5101 cacagcttgg gggtgacgga tcccccaaga agaagaggaa agtctcgagc gactacaaag
     5161 accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaaggctg
     5221 caggaggcgg aggtagcgaa ctgcccacct gcagctgtct tgatcgagtt atacaaaaag
     5281 acaaaggccc atattataca caccttgggg caggaccaag tgttgctgct gtcagggaaa
     5341 tcatggagaa taggtatggt caaaaaggaa acgcaataag gatagaaata gtagtgtaca
     5401 ccggtaaaga agggaaaagc tctcatgggt gtccaattgc taagtgggtt ttaagaagaa
     5461 gcagtgatga agaaaaagtt ctttgtttgg tccggcagcg tacaggccac cactgtccaa
     5521 ctgctgtgat ggtggtgctc atcatggtgt gggatggcat ccctcttcca atggccgacc
     5581 ggctatacac agagctcaca gagaatctaa agtcatacaa tgggcaccct accgacagaa
     5641 gatgcaccct caatgaaaat cgtacctgta catgtcaagg aattgatcca gagacttgtg
     5701 gagcttcatt ctcttttggc tgttcatgga gtatgtactt taatggctgt aagtttggta
     5761 gaagcccaag ccccagaaga tttagaattg atccaagctc tcccttacat gaaaaaaacc
     5821 ttgaagataa cttacagagt ttggctacac gattagctcc aatttataag cagtatgctc
     5881 cagtagctta ccaaaatcag gtggaatatg aaaatgttgc ccgagaatgt cggcttggca
     5941 gcaaggaagg tcgtcccttc tctggggtca ctgcttgcct ggacttctgt gctcatcccc
     6001 acagggacat tcacaacatg aataatggaa gcactgtggt ttgtacctta actcgagaag
     6061 ataaccgctc tttgggtgtt attcctcaag atgagcagct ccatgtgcta cctctttata
     6121 agctttcaga cacagatgag tttggctcca aggaaggaat ggaagccaag atcaaatctg
     6181 gggccatcga ggtcctggca ccccgccgca aaaaaagaac gtgtttcact cagcctgttc
     6241 cccgttctgg aaagaagagg gctgcgatga tgacagaggt tcttgcacat aagataaggg
     6301 cagtggaaaa gaaacctatt ccccgaatca agcggaagaa taactcaaca acaacaaaca
     6361 acagtaagcc ttcgtcactg ccaaccttag ggagtaacac tgagaccgtg caacctgaag
     6421 taaaaagtga aaccgaaccc cattttatct taaaaagttc agacaacact aaaacttatt
     6481 cgctgatgcc atccgctcct cacccagtga aagaggcatc tccaggcttc tcctggtccc
     6541 cgaagactgc ttcagccaca ccagctccac tgaagaatga cgcaacagcc tcatgcgggt
     6601 tttcagaaag aagcagcact ccccactgta cgatgccttc gggaagactc agtggtgcca
     6661 atgcagctgc tgctgatggc cctggcattt cacagcttgg cgaagtggct cctctcccca
     6721 ccctgtctgc tcctgtgatg gagcccctca ttaattctga gccttccact ggtgtgactg
     6781 agccgctaac gcctcatcag ccaaaccacc agccctcctt cctcacctct cctcaagacc
     6841 ttgcctcttc tccaatggaa gaagatgagc agcattctga agcagatgag cctccatcag
     6901 acgaacccct atctgatgac cccctgtcac ctgctgagga gaaattgccc cacattgatg
     6961 agtattggtc agacagtgag cacatctttt tggatgcaaa tattggtggg gtggccatcg
     7021 cacctgctca cggctcggtt ttgattgagt gtgcccggcg agagctgcac gctaccactc
     7081 ctgttgagca ccccaaccgt aatcatccaa cccgcctctc ccttgtcttt taccagcaca
     7141 aaaacctaaa taagccccaa catggttttg aactaaacaa gattaagttt gaggctaaag
     7201 aagctaagaa taagaaaatg aaggcctcag agcaaaaaga ccaggcagct aatgaaggtc
     7261 cagaacagtc ctctgaagta aatgaattga accaaattcc ttctcataaa gcattaacat
     7321 taacccatga caatgttgtc accgtgtccc cttatgctct cacacacgtt gcggggccct
     7381 ataaccattg ggtctgaaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     7441 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     7501 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     7561 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     7621 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgatacc
     7681 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     7741 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctaggg
     7801 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     7861 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7921 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7981 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     8041 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     8101 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8161 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8221 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8281 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8341 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8401 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8461 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8521 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     8581 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8641 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8701 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8761 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8821 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8881 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8941 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9001 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9061 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9121 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9181 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9241 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9301 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9361 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9421 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9481 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9541 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9601 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9661 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9721 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9781 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9841 cacatttccc cgaaaagtgc cacctgacgt cgacgggtat acagacatga taagatacat
     9901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     9961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg
    10021 cgaagaactc tcggcatcta ctctattcct ttgccctcgg acgagtgctg gggcgtcggt
    10081 ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg
    10141 cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct
    10201 gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac
    10261 caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct
    10321 cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg
    10381 cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc
    10441 ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt
    10501 cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga
    10561 cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga
    10621 aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg
    10681 tctggctaag atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag
    10741 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
    10801 aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg
    10861 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
    10921 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
    10981 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
    11041 acgtcgcggt gagttcaggc tttttcatgg tggcgggatg caggtcgaaa ggcccggaga
    11101 tgaggaagag gagaacagcg cggcagacgt gcgcttttga agcgtgcaga atgccgggcc
    11161 tccggaggac cttcgggcgc ccgccccgcc cctgagcccg cccctgagcc cgcccccgga
    11221 cccacccctt ccc
//
