LOCUS       sequence-204464-       10123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-JMJD2A fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10123)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10123)
  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            "Unpublished"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-JMJD2A-Hygro
FEATURES             Location/Qualifiers
     source          1..10123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..26)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     400..423
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        463..842
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        843..1046
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     997..1017
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1088..1107
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1088..1106
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1114..1123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1120..4521,4522..4524,4525..5121,5122,5123..5125,
                     5126..5127,5128..5130,5131..5223)
                     /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(1120..1146,1147..1149,1150..3636,3637..3639,
                     3640..5223)
                     /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             5230..5250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5257..5322
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6409..6426)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6415..6639
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6710..6726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6710..6726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6723..6745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6734..6750
                     /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(6758..6764,6765..6782,6783..6788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6803..6824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6941..6958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7112..7700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7192..7211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7871..8662,8663..8731))
                     /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     8494..8513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8732..8836)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8887..9008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8898..8917)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8952..8971
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9039..10064)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(9039..10064)
                     /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(10082..10123,1..458))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag gaccttcggg cgcccgcccc
       61 gcccctgagc ccgcccctga gcccgccccc ggacccaccc cttcccagcc tctgagccca
      121 gaaagcgaag gagcaaagct gctattggcc gctgccccaa aggcctaccc gcttccattg
      181 ctcagcggtg ctgtccatct gcacgagact agtgagacgt gctacttcca tttgtcacgt
      241 cctgcacgac gcgagctgcg gggcgggggg gaacttcctg actaggggag gagtagaagg
      301 tggcgcgaag gggccaccaa agaacggagc cggttggcgc ctaccggtgg atgtggaatg
      361 tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc agcggggctg ctaaagcgca
      421 tgctccagac tgccttggga aaagcgcctc ccctacccgg ttgacattga ttattgacta
      481 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      541 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      601 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      661 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      721 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      781 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      841 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      901 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      961 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1021 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
     1081 cggccgctaa tacgactcac tatagggaga gccgccacca tggataaaaa gtattctatt
     1141 ggtttagcca tcggcactaa ttccgttgga tgggctgtca taaccgatga atacaaagta
     1201 ccttcaaaga aatttaaggt gttggggaac acagaccgtc attcgattaa aaagaatctt
     1261 atcggtgccc tcctattcga tagtggcgaa acggcagagg cgactcgcct gaaacgaacc
     1321 gctcggagaa ggtatacacg tcgcaagaac cgaatatgtt acttacaaga aatttttagc
     1381 aatgagatgg ccaaagttga cgattctttc tttcaccgtt tggaagagtc cttccttgtc
     1441 gaagaggaca agaaacatga acggcacccc atctttggaa acatagtaga tgaggtggca
     1501 tatcatgaaa agtacccaac gatttatcac ctcagaaaaa agctagttga ctcaactgat
     1561 aaagcggacc tgaggttaat ctacttggct cttgcccata tgataaagtt ccgtgggcac
     1621 tttctcattg agggtgatct aaatccggac aactcggatg tcgacaaact gttcatccag
     1681 ttagtacaaa cctataatca gttgtttgaa gagaacccta taaatgcaag tggcgtggat
     1741 gcgaaggcta ttcttagcgc ccgcctctct aaatcccgac ggctagaaaa cctgatcgca
     1801 caattacccg gagagaagaa aaatgggttg ttcggtaacc ttatagcgct ctcactaggc
     1861 ctgacaccaa attttaagtc gaacttcgac ttagctgaag atgccaaatt gcagcttagt
     1921 aaggacacgt acgatgacga tctcgacaat ctactggcac aaattggaga tcagtatgcg
     1981 gacttatttt tggctgccaa aaaccttagc gatgcaatcc tcctatctga catactgaga
     2041 gttaatactg agattaccaa ggcgccgtta tccgcttcaa tgatcaaaag gtacgatgaa
     2101 catcaccaag acttgacact tctcaaggcc ctagtccgtc agcaactgcc tgagaaatat
     2161 aaggaaatat tctttgatca gtcgaaaaac gggtacgcag gttatattga cggcggagcg
     2221 agtcaagagg aattctacaa gtttatcaaa cccatattag agaagatgga tgggacggaa
     2281 gagttgcttg taaaactcaa tcgcgaagat ctactgcgaa agcagcggac tttcgacaac
     2341 ggtagcattc cacatcaaat ccacttaggc gaattgcatg ctatacttag aaggcaggag
     2401 gatttttatc cgttcctcaa agacaatcgt gaaaagattg agaaaatcct aacctttcgc
     2461 ataccttact atgtgggacc cctggcccga gggaactctc ggttcgcatg gatgacaaga
     2521 aagtccgaag aaacgattac tccatggaat tttgaggaag ttgtcgataa aggtgcgtca
     2581 gctcaatcgt tcatcgagag gatgaccaac tttgacaaga atttaccgaa cgaaaaagta
     2641 ttgcctaagc acagtttact ttacgagtat ttcacagtgt acaatgaact cacgaaagtt
     2701 aagtatgtca ctgagggcat gcgtaaaccc gcctttctaa gcggagaaca gaagaaagca
     2761 atagtagatc tgttattcaa gaccaaccgc aaagtgacag ttaagcaatt gaaagaggac
     2821 tactttaaga aaattgaatg cttcgattct gtcgagatct ccggggtaga agatcgattt
     2881 aatgcgtcac ttggtacgta tcatgacctc ctaaagataa ttaaagataa ggacttcctg
     2941 gataacgaag agaatgaaga tatcttagaa gatatagtgt tgactcttac cctctttgaa
     3001 gatcgggaaa tgattgagga aagactaaaa acatacgctc acctgttcga cgataaggtt
     3061 atgaaacagt taaagaggcg tcgctatacg ggctggggac gattgtcgcg gaaacttatc
     3121 aacgggataa gagacaagca aagtggtaaa actattctcg attttctaaa gagcgacggc
     3181 ttcgccaata ggaactttat gcagctgatc catgatgact ctttaacctt caaagaggat
     3241 atacaaaagg cacaggtttc cggacaaggg gactcattgc acgaacatat tgcgaatctt
     3301 gctggttcgc cagccatcaa aaagggcata ctccagacag tcaaagtagt ggatgagcta
     3361 gttaaggtca tgggacgtca caaaccggaa aacattgtaa tcgagatggc acgcgaaaat
     3421 caaacgactc agaaggggca aaaaaacagt cgagagcgga tgaagagaat agaagagggt
     3481 attaaagaac tgggcagcca gatcttaaag gagcatcctg tggaaaatac ccaattgcag
     3541 aacgagaaac tttacctcta ttacctacaa aatggaaggg acatgtatgt tgatcaggaa
     3601 ctggacataa accgtttatc tgattacgac gtcgatgcca ttgtacccca atcctttttg
     3661 aaggacgatt caatcgacaa taaagtgctt acacgctcgg ataagaaccg agggaaaagt
     3721 gacaatgttc caagcgagga agtcgtaaag aaaatgaaga actattggcg gcagctccta
     3781 aatgcgaaac tgataacgca aagaaagttc gataacttaa ctaaagctga gaggggtggc
     3841 ttgtctgaac ttgacaaggc cggatttatt aaacgtcagc tcgtggaaac ccgccaaatc
     3901 acaaagcatg ttgcacagat actagattcc cgaatgaata cgaaatacga cgagaacgat
     3961 aagctgattc gggaagtcaa agtaatcact ttaaagtcaa aattggtgtc ggacttcaga
     4021 aaggattttc aattctataa agttagggag ataaataact accaccatgc gcacgacgct
     4081 tatcttaatg ccgtcgtagg gaccgcactc attaagaaat acccgaagct agaaagtgag
     4141 tttgtgtatg gtgattacaa agtttatgac gtccgtaaga tgatcgcgaa aagcgaacag
     4201 gagataggca aggctacagc caaatacttc ttttattcta acattatgaa tttctttaag
     4261 acggaaatca ctctggcaaa cggagagata cgcaaacgac ctttaattga aaccaatggg
     4321 gagacaggtg aaatcgtatg ggataagggc cgggacttcg cgacggtgag aaaagttttg
     4381 tccatgcccc aagtcaacat agtaaagaaa actgaggtgc agaccggagg gttttcaaag
     4441 gaatcgattc ttccaaaaag gaatagtgat aagctcatcg ctcgtaaaaa ggactgggac
     4501 ccgaaaaagt acggtggctt cgatagccct acagttgcct attctgtcct agtagtggca
     4561 aaagttgaga agggaaaatc caagaaactg aagtcagtca aagaattatt ggggataacg
     4621 attatggagc gctcgtcttt tgaaaagaac cccatcgact tccttgaggc gaaaggttac
     4681 aaggaagtaa aaaaggatct cataattaaa ctaccaaagt atagtctgtt tgagttagaa
     4741 aatggccgaa aacggatgtt ggctagcgcc ggagagcttc aaaaggggaa cgaactcgca
     4801 ctaccgtcta aatacgtgaa tttcctgtat ttagcgtccc attacgagaa gttgaaaggt
     4861 tcacctgaag ataacgaaca gaagcaactt tttgttgagc agcacaaaca ttatctcgac
     4921 gaaatcatag agcaaatttc ggaattcagt aagagagtca tcctagctga tgccaatctg
     4981 gacaaagtat taagcgcata caacaagcac agggataaac ccatacgtga gcaggcggaa
     5041 aatattatcc atttgtttac tcttaccaac ctcggcgctc cagccgcatt caagtatttt
     5101 gacacaacga tagatcgcaa acgatacact tctaccaagg aggtgctaga cgcgacactg
     5161 attcaccaat ccatcacggg attatatgaa actcggatag atttgtcaca gcttgggggt
     5221 gacggatccc ccaagaagaa gaggaaagtc tcgagcgact acaaagacca tgacggtgat
     5281 tataaagatc atgacatcga ttacaaggat gacgatgaca aggctgcagg aggcggaggt
     5341 agcgcttctg agtctgaaac tctgaatccc agtgctagga taatgacctt ttatccaact
     5401 atggaagagt tccgaaactt cagtagatac attgcctaca ttgaatccca aggagctcat
     5461 cgggcagggc tagccaaggt tgttcctcca aaagagtgga agccacgagc atcctatgat
     5521 gacattgatg atttggtcat tcctgccccc attcaacagc tggtgacggg gcagtctggc
     5581 ctctttactc agtacaacat acagaagaaa gccatgactg ttcgagagtt ccgcaagata
     5641 gccaatagcg ataagtactg taccccacgc tatagtgagt ttgaagagct cgagcggaaa
     5701 tactggaaaa atcttacatt caatcctcca atctatggtg cagatgtgaa tggtaccctc
     5761 tatgaaaagc atgttgatga gtggaatatt ggccggctga gaacaatcct ggacttggtg
     5821 gaaaaggaga gtgggatcac cattgagggt gtgaacaccc catacctgta ctttggcatg
     5881 tggaagacat cctttgcttg gcacactgaa gacatggacc tctacagcat caactacctg
     5941 cactttggag aaccaaagtc ctggtactct gttccacctg agcatggaaa gcggttggaa
     6001 cgcctcgcca aaggcttttt cccaggaagt gctcaaagct gtgaggcatt tctccgccac
     6061 aagatgaccc tgatttcccc gttaatgctg aagaaatatg gaattccctt tgacaaggtg
     6121 actcaagagg ctggagagtt tatgatcact ttcccttatg gttaccatgc cggctttaac
     6181 catggtttta actgtgcgga gtctaccaat tttgctaccc gtcggtggat tgagtacggc
     6241 aagcaagctg tgctgtgctc ctgtagaaag gacatggtga agatctccat ggatgtgttt
     6301 gtgagaaagt tccagccaga aaggtacaaa ctttggaaag ctgggaagga caacacagtt
     6361 attgaccata ctctgcccac gccagaagca tgaaaacccg ctgatcagcc tcgactgtgc
     6421 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     6481 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     6541 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     6601 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     6661 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6721 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6781 agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6841 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6901 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac gggtatacag
     8881 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     8941 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     9001 aacaagttgg ggtgggcgaa gaactctcgg catctactct attcctttgc cctcggacga
     9061 gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg
     9121 gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt
     9181 gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga
     9241 tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc
     9301 tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc
     9361 cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca
     9421 atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc
     9481 acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc
     9541 gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca
     9601 gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat
     9661 gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg
     9721 accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt
     9781 gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc
     9841 ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca
     9901 tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca
     9961 cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc
    10021 agaaacttct cgacagacgt cgcggtgagt tcaggctttt tcatggtggc gggatgcagg
    10081 tcgaaaggcc cggagatgag gaagaggaga acagcgcggc aga
//
