LOCUS       sequence-364146-       13033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VPR activation domain coupled to catalytically inactive dCas9 for 
            localization; contains a GFP tetramer fpr labeling and as a spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13033)
  AUTHORS   Trojanowski J, Frank L, Rademacher A, Mucke N, Grigaitis P, Rippe K
  TITLE     Transcription activation is enhanced by multivalent interactions 
            independent of phase separation.
  JOURNAL   Mol Cell. 2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017.
  PUBMED    35537448
REFERENCE   2  (bases 1 to 13033)
  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: "Mol Cell. 
            2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017."
COMMENT     Sequence Label: NLS-dCas9-HA-NLS-GFP4-VPR
FEATURES             Location/Qualifiers
     source          1..13033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             645..665
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      714..723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(720..746,747..749,750..3236,3237..3239,3240..4823)
                     /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             720..4823
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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             4827..4853
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4860..4880
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4938..4947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4944..4946,4947..4949,4950..5660)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4944..4946,4947..4949,4950..5660)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      5730..5739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5736..5738,5739..5741,5742..6452)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5736..5738,5739..5741,5742..6452)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      6522..6531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6528..6530,6531..6533,6534..7244)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6528..6530,6531..6533,6534..7244)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      7314..7323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7320..7322,7323..7325,7326..8036)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7320..7322,7323..7325,7326..8036)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             8070..8090
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8157..8306
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             8331..8351
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8781..9137
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             9156..9725
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     polyA_signal    9860..9981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9897..9916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9951..9970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(9988..10443)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10125..10146)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10337..10356
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        10470..10574
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        10576..10767
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(10599..10619)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      10764..10897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             10946..11740
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             10946..11740
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(11000..11019)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11610..11629
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(11928..11947)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11972..12019
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      12348..12936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12837..12856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc catgcattag ttattaatag taatcaatta ctgggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agcgctaccg gactcagatc tcgaggccac catgcccaag aagaagcgca
      661 aggtgggacg cgtctgcagg atatcaagct tgcggtaccg cgggcccggg atcgccacca
      721 tggacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga
      781 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
      841 acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa acagccgagg
      901 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
      961 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     1021 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     1081 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     1141 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     1201 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     1261 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     1321 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     1381 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggcaacc
     1441 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     1501 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1561 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1621 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     1681 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     1741 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     1801 gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     1861 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     1921 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     1981 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     2041 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     2101 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     2161 tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     2221 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     2281 acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     2341 gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     2401 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     2461 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     2521 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     2581 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     2641 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     2701 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     2761 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     2821 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     2881 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     2941 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     3001 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     3061 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     3121 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     3181 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggacgcta
     3241 tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg actcggagcg
     3301 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     3361 actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc gacaatctga
     3421 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     3481 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     3541 ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc ctgaagtcca
     3601 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     3661 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     3721 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     3781 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     3841 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     3901 ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc cgggactttg
     3961 ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     4021 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac aagctgatcg
     4081 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     4141 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     4201 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     4261 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     4321 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     4381 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     4441 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     4501 agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     4561 tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac agagacaagc
     4621 ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     4681 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     4741 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     4801 acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat gcaccggtcc
     4861 caaaaaagaa aagaaaagta ccggtcgcca cccgaattct gcagtcgacg gtaccgcggg
     4921 cccgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     4981 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     5041 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     5101 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     5161 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     5221 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     5281 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     5341 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     5401 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     5461 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     5521 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     5581 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     5641 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag
     5701 tcgacggtac cgcgggcccg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     5761 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     5821 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     5881 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     5941 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     6001 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     6061 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     6121 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     6181 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     6241 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     6301 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     6361 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     6421 ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt
     6481 cgaattctgc agtcgacggt accgcgggcc cgctaccggt cgccaccatg gtgagcaagg
     6541 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     6601 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     6661 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     6721 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     6781 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     6841 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     6901 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     6961 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     7021 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     7081 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     7141 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     7201 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc
     7261 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgctaccg gtcgccacca
     7321 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     7381 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     7441 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     7501 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     7561 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     7621 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     7681 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     7741 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     7801 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     7861 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     7921 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     7981 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagacta
     8041 gtcgcagggg gcgtcgactg ggtacccctc ctaagaagaa gaggaaggtt gcccccacca
     8101 tggataaaaa gaagagaaag gcccccccga ccgatgtcag cctgggggac gagctcgacg
     8161 cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat tttgaccttg
     8221 acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc agtgacgccc
     8281 ttgatgattt cgacctggac atgctgatta actctagaag ttccggatct ccgaaaaaga
     8341 aacgcaaagt tggtagccag tacctgcccg acaccgacga ccggcaccgg atcgaggaaa
     8401 agcggaagcg gacctacgag acattcaaga gcatcatgaa gaagtccccc ttcagcggcc
     8461 ccaccgaccc tagacctcca cctagaagaa tcgccgtgcc cagcagatcc agcgccagcg
     8521 tgccaaaacc tgccccccag ccttacccct tcaccagcag cctgagcacc atcaactacg
     8581 acgagttccc taccatggtg ttccccagcg gccagatctc tcaggcctct gctctggctc
     8641 cagcccctcc tcaggtgctg cctcaggctc ctgctcctgc accagctcca gccatggtgt
     8701 ctgcactggc tcaggcacca gcacccgtgc ctgtgctggc tcctggacct ccacaggctg
     8761 tggctccacc agcccctaaa cctacacagg ccggcgaggg cacactgtct gaagctctgc
     8821 tgcagctgca gttcgacgac gaggatctgg gagccctgct gggaaacagc accgatcctg
     8881 ccgtgttcac cgacctggcc agcgtggaca acagcgagtt ccagcagctg ctgaaccagg
     8941 gcatccctgt ggcccctcac accaccgagc ccatgctgat ggaatacccc gaggccatca
     9001 cccggctcgt gacaggcgct cagaggcctc ctgatccagc tcctgcccct ctgggagcac
     9061 caggcctgcc taatggactg ctgtctggcg acgaggactt cagctctatc gccgatatgg
     9121 atttctcagc cttgctgggc tctggcagcg gcagccggga ttccagggaa gggatgtttt
     9181 tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt tgagggccgc gaggtgtgcc
     9241 agccaaaacg aatccggcca tttcatcctc caggaagtcc atgggccaac cgcccactcc
     9301 ccgccagcct cgcaccaaca ccaaccggtc cagtacatga gccagtcggg tcactgaccc
     9361 cggcaccagt ccctcagcca ctggatccag cgcccgcagt gactcccgag gccagtcacc
     9421 tgttggagga tcccgatgaa gagacgagcc aggctgtcaa agcccttcgg gagatggccg
     9481 atactgtgat tccccagaag gaagaggctg caatctgtgg ccaaatggac ctttcccatc
     9541 cgcccccaag gggccatctg gatgagctga caaccacact tgagtccatg accgaggatc
     9601 tgaacctgga ctcacccctg accccggaat tgaacgagat tctggatacc ttcctgaacg
     9661 acgagtgcct cttgcatgcc atgcatatca gcacaggact gtccatcttc gacacatctc
     9721 tgttttaaaa cgttcagagc ggccgcgact ctagatcata atcagccata ccacatttgt
     9781 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     9841 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     9901 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     9961 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
    10021 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
    10081 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
    10141 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
    10201 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
    10261 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
    10321 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
    10381 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
    10441 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
    10501 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
    10561 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
    10621 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
    10681 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
    10741 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccattctccg
    10801 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
    10861 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
    10921 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
    10981 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
    11041 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
    11101 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
    11161 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
    11221 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
    11281 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
    11341 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
    11401 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
    11461 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
    11521 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
    11581 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
    11641 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
    11701 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
    11761 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
    11821 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
    11881 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
    11941 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
    12001 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
    12061 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
    12121 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
    12181 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
    12241 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
    12301 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
    12361 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
    12421 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
    12481 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
    12541 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
    12601 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
    12661 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
    12721 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
    12781 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
    12841 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
    12901 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
    12961 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
    13021 tctgtggata acc
//
