LOCUS       sequence-204141-       11233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-dTET1 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  JOURNAL   Unpublished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            "Unpublished"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-dTET1-H1672Y-D1674A-Hygro
FEATURES             Location/Qualifiers
     source          1..11233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..43)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     417..440
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        480..859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        860..1063
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1014..1034
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1105..1124
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1105..1123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1131..1140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1137..4538,4539..4541,4542..5138,5139,5140..5142,
                     5143..5144,5145..5147,5148..5240)
                     /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(1137..1163,1164..1166,1167..3653,3654..3656,
                     3657..5240)
                     /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             5247..5267
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5274..5339
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7536..7553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7542..7766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7837..7853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7837..7853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7850..7872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7861..7877
                     /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(7885..7891,7892..7909,7910..7915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7930..7951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8068..8085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8239..8827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8319..8338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8998..9789,9790..9858))
                     /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     9621..9640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9859..9963)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10014..10135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10025..10044)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10079..10098
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(10166..11191)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(10166..11191)
                     /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(11209..11233,1..475))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 gagaacagcg cggcagacgt gcgcttttga agcgtgcaga atgccgggcc tccggaggac
       61 cttcgggcgc ccgccccgcc cctgagcccg cccctgagcc cgcccccgga cccacccctt
      121 cccagcctct gagcccagaa agcgaaggag caaagctgct attggccgct gccccaaagg
      181 cctacccgct tccattgctc agcggtgctg tccatctgca cgagactagt gagacgtgct
      241 acttccattt gtcacgtcct gcacgacgcg agctgcgggg cgggggggaa cttcctgact
      301 aggggaggag tagaaggtgg cgcgaagggg ccaccaaaga acggagccgg ttggcgccta
      361 ccggtggatg tggaatgtgt gcgaggccag aggccacttg tgtagcgcca agtgcccagc
      421 ggggctgcta aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc tacccggttg
      481 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      541 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      601 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      661 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      721 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      781 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      841 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      901 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      961 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     1021 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     1081 gatccgctag agatccgcgg ccgctaatac gactcactat agggagagcc gccaccatgg
     1141 ataaaaagta ttctattggt ttagccatcg gcactaattc cgttggatgg gctgtcataa
     1201 ccgatgaata caaagtacct tcaaagaaat ttaaggtgtt ggggaacaca gaccgtcatt
     1261 cgattaaaaa gaatcttatc ggtgccctcc tattcgatag tggcgaaacg gcagaggcga
     1321 ctcgcctgaa acgaaccgct cggagaaggt atacacgtcg caagaaccga atatgttact
     1381 tacaagaaat ttttagcaat gagatggcca aagttgacga ttctttcttt caccgtttgg
     1441 aagagtcctt ccttgtcgaa gaggacaaga aacatgaacg gcaccccatc tttggaaaca
     1501 tagtagatga ggtggcatat catgaaaagt acccaacgat ttatcacctc agaaaaaagc
     1561 tagttgactc aactgataaa gcggacctga ggttaatcta cttggctctt gcccatatga
     1621 taaagttccg tgggcacttt ctcattgagg gtgatctaaa tccggacaac tcggatgtcg
     1681 acaaactgtt catccagtta gtacaaacct ataatcagtt gtttgaagag aaccctataa
     1741 atgcaagtgg cgtggatgcg aaggctattc ttagcgcccg cctctctaaa tcccgacggc
     1801 tagaaaacct gatcgcacaa ttacccggag agaagaaaaa tgggttgttc ggtaacctta
     1861 tagcgctctc actaggcctg acaccaaatt ttaagtcgaa cttcgactta gctgaagatg
     1921 ccaaattgca gcttagtaag gacacgtacg atgacgatct cgacaatcta ctggcacaaa
     1981 ttggagatca gtatgcggac ttatttttgg ctgccaaaaa ccttagcgat gcaatcctcc
     2041 tatctgacat actgagagtt aatactgaga ttaccaaggc gccgttatcc gcttcaatga
     2101 tcaaaaggta cgatgaacat caccaagact tgacacttct caaggcccta gtccgtcagc
     2161 aactgcctga gaaatataag gaaatattct ttgatcagtc gaaaaacggg tacgcaggtt
     2221 atattgacgg cggagcgagt caagaggaat tctacaagtt tatcaaaccc atattagaga
     2281 agatggatgg gacggaagag ttgcttgtaa aactcaatcg cgaagatcta ctgcgaaagc
     2341 agcggacttt cgacaacggt agcattccac atcaaatcca cttaggcgaa ttgcatgcta
     2401 tacttagaag gcaggaggat ttttatccgt tcctcaaaga caatcgtgaa aagattgaga
     2461 aaatcctaac ctttcgcata ccttactatg tgggacccct ggcccgaggg aactctcggt
     2521 tcgcatggat gacaagaaag tccgaagaaa cgattactcc atggaatttt gaggaagttg
     2581 tcgataaagg tgcgtcagct caatcgttca tcgagaggat gaccaacttt gacaagaatt
     2641 taccgaacga aaaagtattg cctaagcaca gtttacttta cgagtatttc acagtgtaca
     2701 atgaactcac gaaagttaag tatgtcactg agggcatgcg taaacccgcc tttctaagcg
     2761 gagaacagaa gaaagcaata gtagatctgt tattcaagac caaccgcaaa gtgacagtta
     2821 agcaattgaa agaggactac tttaagaaaa ttgaatgctt cgattctgtc gagatctccg
     2881 gggtagaaga tcgatttaat gcgtcacttg gtacgtatca tgacctccta aagataatta
     2941 aagataagga cttcctggat aacgaagaga atgaagatat cttagaagat atagtgttga
     3001 ctcttaccct ctttgaagat cgggaaatga ttgaggaaag actaaaaaca tacgctcacc
     3061 tgttcgacga taaggttatg aaacagttaa agaggcgtcg ctatacgggc tggggacgat
     3121 tgtcgcggaa acttatcaac gggataagag acaagcaaag tggtaaaact attctcgatt
     3181 ttctaaagag cgacggcttc gccaatagga actttatgca gctgatccat gatgactctt
     3241 taaccttcaa agaggatata caaaaggcac aggtttccgg acaaggggac tcattgcacg
     3301 aacatattgc gaatcttgct ggttcgccag ccatcaaaaa gggcatactc cagacagtca
     3361 aagtagtgga tgagctagtt aaggtcatgg gacgtcacaa accggaaaac attgtaatcg
     3421 agatggcacg cgaaaatcaa acgactcaga aggggcaaaa aaacagtcga gagcggatga
     3481 agagaataga agagggtatt aaagaactgg gcagccagat cttaaaggag catcctgtgg
     3541 aaaataccca attgcagaac gagaaacttt acctctatta cctacaaaat ggaagggaca
     3601 tgtatgttga tcaggaactg gacataaacc gtttatctga ttacgacgtc gatgccattg
     3661 taccccaatc ctttttgaag gacgattcaa tcgacaataa agtgcttaca cgctcggata
     3721 agaaccgagg gaaaagtgac aatgttccaa gcgaggaagt cgtaaagaaa atgaagaact
     3781 attggcggca gctcctaaat gcgaaactga taacgcaaag aaagttcgat aacttaacta
     3841 aagctgagag gggtggcttg tctgaacttg acaaggccgg atttattaaa cgtcagctcg
     3901 tggaaacccg ccaaatcaca aagcatgttg cacagatact agattcccga atgaatacga
     3961 aatacgacga gaacgataag ctgattcggg aagtcaaagt aatcacttta aagtcaaaat
     4021 tggtgtcgga cttcagaaag gattttcaat tctataaagt tagggagata aataactacc
     4081 accatgcgca cgacgcttat cttaatgccg tcgtagggac cgcactcatt aagaaatacc
     4141 cgaagctaga aagtgagttt gtgtatggtg attacaaagt ttatgacgtc cgtaagatga
     4201 tcgcgaaaag cgaacaggag ataggcaagg ctacagccaa atacttcttt tattctaaca
     4261 ttatgaattt ctttaagacg gaaatcactc tggcaaacgg agagatacgc aaacgacctt
     4321 taattgaaac caatggggag acaggtgaaa tcgtatggga taagggccgg gacttcgcga
     4381 cggtgagaaa agttttgtcc atgccccaag tcaacatagt aaagaaaact gaggtgcaga
     4441 ccggagggtt ttcaaaggaa tcgattcttc caaaaaggaa tagtgataag ctcatcgctc
     4501 gtaaaaagga ctgggacccg aaaaagtacg gtggcttcga tagccctaca gttgcctatt
     4561 ctgtcctagt agtggcaaaa gttgagaagg gaaaatccaa gaaactgaag tcagtcaaag
     4621 aattattggg gataacgatt atggagcgct cgtcttttga aaagaacccc atcgacttcc
     4681 ttgaggcgaa aggttacaag gaagtaaaaa aggatctcat aattaaacta ccaaagtata
     4741 gtctgtttga gttagaaaat ggccgaaaac ggatgttggc tagcgccgga gagcttcaaa
     4801 aggggaacga actcgcacta ccgtctaaat acgtgaattt cctgtattta gcgtcccatt
     4861 acgagaagtt gaaaggttca cctgaagata acgaacagaa gcaacttttt gttgagcagc
     4921 acaaacatta tctcgacgaa atcatagagc aaatttcgga attcagtaag agagtcatcc
     4981 tagctgatgc caatctggac aaagtattaa gcgcatacaa caagcacagg gataaaccca
     5041 tacgtgagca ggcggaaaat attatccatt tgtttactct taccaacctc ggcgctccag
     5101 ccgcattcaa gtattttgac acaacgatag atcgcaaacg atacacttct accaaggagg
     5161 tgctagacgc gacactgatt caccaatcca tcacgggatt atatgaaact cggatagatt
     5221 tgtcacagct tgggggtgac ggatccccca agaagaagag gaaagtctcg agcgactaca
     5281 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg
     5341 ctgcaggagg cggaggtagc gaactgccca cctgcagctg tcttgatcga gttatacaaa
     5401 aagacaaagg cccatattat acacaccttg gggcaggacc aagtgttgct gctgtcaggg
     5461 aaatcatgga gaataggtat ggtcaaaaag gaaacgcaat aaggatagaa atagtagtgt
     5521 acaccggtaa agaagggaaa agctctcatg ggtgtccaat tgctaagtgg gttttaagaa
     5581 gaagcagtga tgaagaaaaa gttctttgtt tggtccggca gcgtacaggc caccactgtc
     5641 caactgctgt gatggtggtg ctcatcatgg tgtgggatgg catccctctt ccaatggccg
     5701 accggctata cacagagctc acagagaatc taaagtcata caatgggcac cctaccgaca
     5761 gaagatgcac cctcaatgaa aatcgtacct gtacatgtca aggaattgat ccagagactt
     5821 gtggagcttc attctctttt ggctgttcat ggagtatgta ctttaatggc tgtaagtttg
     5881 gtagaagccc aagccccaga agatttagaa ttgatccaag ctctccctta catgaaaaaa
     5941 accttgaaga taacttacag agtttggcta cacgattagc tccaatttat aagcagtatg
     6001 ctccagtagc ttaccaaaat caggtggaat atgaaaatgt tgcccgagaa tgtcggcttg
     6061 gcagcaagga aggtcgtccc ttctctgggg tcactgcttg cctggacttc tgtgctcatc
     6121 cctaccgggc cattcacaac atgaataatg gaagcactgt ggtttgtacc ttaactcgag
     6181 aagataaccg ctctttgggt gttattcctc aagatgagca gctccatgtg ctacctcttt
     6241 ataagctttc agacacagat gagtttggct ccaaggaagg aatggaagcc aagatcaaat
     6301 ctggggccat cgaggtcctg gcaccccgcc gcaaaaaaag aacgtgtttc actcagcctg
     6361 ttccccgttc tggaaagaag agggctgcga tgatgacaga ggttcttgca cataagataa
     6421 gggcagtgga aaagaaacct attccccgaa tcaagcggaa gaataactca acaacaacaa
     6481 acaacagtaa gccttcgtca ctgccaacct tagggagtaa cactgagacc gtgcaacctg
     6541 aagtaaaaag tgaaaccgaa ccccatttta tcttaaaaag ttcagacaac actaaaactt
     6601 attcgctgat gccatccgct cctcacccag tgaaagaggc atctccaggc ttctcctggt
     6661 ccccgaagac tgcttcagcc acaccagctc cactgaagaa tgacgcaaca gcctcatgcg
     6721 ggttttcaga aagaagcagc actccccact gtacgatgcc ttcgggaaga ctcagtggtg
     6781 ccaatgcagc tgctgctgat ggccctggca tttcacagct tggcgaagtg gctcctctcc
     6841 ccaccctgtc tgctcctgtg atggagcccc tcattaattc tgagccttcc actggtgtga
     6901 ctgagccgct aacgcctcat cagccaaacc accagccctc cttcctcacc tctcctcaag
     6961 accttgcctc ttctccaatg gaagaagatg agcagcattc tgaagcagat gagcctccat
     7021 cagacgaacc cctatctgat gaccccctgt cacctgctga ggagaaattg ccccacattg
     7081 atgagtattg gtcagacagt gagcacatct ttttggatgc aaatattggt ggggtggcca
     7141 tcgcacctgc tcacggctcg gttttgattg agtgtgcccg gcgagagctg cacgctacca
     7201 ctcctgttga gcaccccaac cgtaatcatc caacccgcct ctcccttgtc ttttaccagc
     7261 acaaaaacct aaataagccc caacatggtt ttgaactaaa caagattaag tttgaggcta
     7321 aagaagctaa gaataagaaa atgaaggcct cagagcaaaa agaccaggca gctaatgaag
     7381 gtccagaaca gtcctctgaa gtaaatgaat tgaaccaaat tccttctcat aaagcattaa
     7441 cattaaccca tgacaatgtt gtcaccgtgt ccccttatgc tctcacacac gttgcggggc
     7501 cctataacca ttgggtctga aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     7561 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     7621 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     7681 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     7741 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     7801 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     7921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     8041 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8101 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8161 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8221 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8281 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8341 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8401 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8461 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8521 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8581 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8641 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8701 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8761 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8821 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8881 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8941 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     9001 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     9061 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     9121 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     9181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9961 gcgcacattt ccccgaaaag tgccacctga cgtcgacggg tatacagaca tgataagata
    10021 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
    10081 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt
    10141 gggcgaagaa ctctcggcat ctactctatt cctttgccct cggacgagtg ctggggcgtc
    10201 ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc
    10261 gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac
    10321 cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa
    10381 gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc
    10441 gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt
    10501 tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta
    10561 tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga
    10621 cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac
    10681 tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata
    10741 tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc
    10801 tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc ggctgcagaa
    10861 cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga
    10921 tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga atcgggagcg
    10981 cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat
    11041 ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga gattcttcgc
    11101 cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga aacttctcga
    11161 cagacgtcgc ggtgagttca ggctttttca tggtggcggg atgcaggtcg aaaggcccgg
    11221 agatgaggaa gag
//
