LOCUS       sequence-262414-       12555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expressing effector domains fused to dCas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12555)
  AUTHORS   Goubert D, Koncz M, Kiss A, Rots MG
  TITLE     Establishment of Cell Lines Stably Expressing dCas9-Fusions to 
            Address Kinetics of Epigenetic Editing.
  JOURNAL   Methods Mol Biol. 2018;1767:395-415. doi: 
            10.1007/978-1-4939-7774-1_22.
  PUBMED    29524148
REFERENCE   2  (bases 1 to 12555)
  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: "Methods Mol
            Biol."; date: "2018"; volume: "1767:395-415. doi"; pages: " 
            10.1007/978-1-4939-7774-1_22"
COMMENT     Sequence Label: pHAGE EF1-alpha dCas9-NED
FEATURES             Location/Qualifiers
     source          1..12555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2220..3398
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2451..3389
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3346..3366
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3433..3457
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3461..3470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3467..3493,3494..3496,3497..5980,5981..5986,5987..6052,
                     6053..6055,6056..7570)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /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(3467..3493,3494..3496,3497..5983,5984..5986,
                     5987..7570)
                     /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             3467..7570
                     /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             7583..7603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7604..7693
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             7754..7780
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        7813..8312
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7848..7871)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8245..8264
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8333..8932
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8333..8352)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8829..8849
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8945..9533
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8998..9018)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9416..9427)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9608..9841
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9960..9978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10046..10150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10151..10219,10220..11011)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10369..10388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11182..11770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11671..11690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11924..11941
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12181..12202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12217..12222,12223..12240,12241..12247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12255..12271
                     /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)."
     primer_bind     12260..12282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12279..12295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12279..12295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12345..12364
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactacgatg
     2221 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttgtggg
     2281 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     2341 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     2401 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     2461 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     2521 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     2581 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     2641 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     2701 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     2761 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     2821 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     2881 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     2941 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     3001 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     3061 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     3121 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     3181 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     3241 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     3301 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     3361 tggttcaaag tttttttctt ccatttcagg tgtcgtgaac tagtccagtg tggtggaatt
     3421 ctgcagatat caacaagttt gtacaaaaaa gcaggctggc gccaccatgg acaagaagta
     3481 ctccattggg ctcgctatcg gtaccaacag cgtcggctgg gccgtcatta cggacgagta
     3541 caaggtgccg agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa
     3601 gaacctcatt ggagccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa
     3661 aagaacagca cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat
     3721 ctttagtaat gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt
     3781 tttggtggag gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga
     3841 ggtggcgtac catgaaaagt acccaaccat atatcatctg aggaagaagc tggtagacag
     3901 tactgataag gctgacttgc ggttgatcta tctcgcgctg gcgcacatga tcaaatttcg
     3961 gggacacttc ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt
     4021 tatccaactg gttcagactt acaatcagct tttcgaggag aacccgatca acgcatccgg
     4081 cgttgacgcc aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct
     4141 catcgcacag ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc
     4201 actcgggctg acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagctgca
     4261 actgagcaaa gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca
     4321 gtacgcagac ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat
     4381 tctgcgagtg aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta
     4441 tgatgagcac caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga
     4501 gaagtacaag gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg
     4561 cggagcaagc caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg
     4621 caccgaggag ctgctggtaa agctgaacag agaagatctg ttgcgcaaac agcgcacttt
     4681 cgacaatgga agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg
     4741 gcaagaggat ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac
     4801 atttcggata ccctactatg taggccccct cgctcgggga aattccagat tcgcgtggat
     4861 gactcgcaaa tcagaagaga ccatcactcc ctggaacttc gaggaagtcg tggataaggg
     4921 ggcctctgcc cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga
     4981 aaaggtgctt cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac
     5041 caaggtcaaa tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa
     5101 gaaagctatc gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa
     5161 agaagactat ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga
     5221 tcgcttcaac gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga
     5281 cttcctggac aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt
     5341 gtttgaagat agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga
     5401 caaagtcatg aaacagctca agagacgccg atatacagga tgggggcggc tgtcaagaaa
     5461 actgatcaat ggcatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc
     5521 cgatggattt gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa
     5581 ggaggacatc cagaaagcac aagtttctgg ccagggggac agtcttcacg agcacatcgc
     5641 taatcttgca ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga
     5701 tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg
     5761 agagaaccaa actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga
     5821 agagggtata aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca
     5881 gcttcagaat gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga
     5941 tcaggaactg gacatcaacc ggttgtccga ctacgacgtg gatgctatcg tgccccaaag
     6001 ctttctcaaa gatgattcta ttgataataa agtgttgaca agatccgata aaaatagagg
     6061 gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca
     6121 gctgctgaac gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg
     6181 aggtggcctg tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg
     6241 ccagatcacc aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga
     6301 aaatgacaaa ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtctcaga
     6361 tttcagaaag gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca
     6421 tgatgcctac ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagctgga
     6481 atctgaattt gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc
     6541 tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt
     6601 tttcaagacc gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac
     6661 aaacggagaa acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccgcaa
     6721 ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt
     6781 ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga
     6841 ttgggacccc aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt
     6901 tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg
     6961 catcacaatc atggagcgat ccagcttcga gaaaaacccc atcgactttc tcgaagcgaa
     7021 aggatataaa gaggtcaaaa aagacctcat cattaagctg cccaagtact ctctctttga
     7081 gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga
     7141 gctggcactg ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct
     7201 caaagggtct cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta
     7261 ccttgatgag atcatcgagc aaataagcga gttctccaaa agagtgatcc tcgccgacgc
     7321 taacctcgat aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca
     7381 ggcagaaaac attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa
     7441 gtacttcgac accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc
     7501 cacactgatt catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct
     7561 cggtggagac ggcaccggcg ggcccaagaa gaagaggaag gtatacccat acgatgttcc
     7621 tgactatgcg ggctatccct atgacgtccc ggactatgca ggatcgtatc cttatgacgt
     7681 tccagattac gctggatcca cacgcgttaa gcgatcgcgc ggccgctcga gtctagaggg
     7741 cccgcggttc gaatacccat acgacgtccc agactacgct tagtaatgat taattaaact
     7801 agaaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     7861 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     7921 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     7981 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     8041 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     8101 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     8161 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     8221 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     8281 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga
     8341 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     8401 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     8461 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     8521 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     8581 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     8641 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     8701 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     8761 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     8821 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     8881 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagcgataga
     8941 tcctaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     9001 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     9061 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     9121 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     9181 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     9241 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     9301 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     9361 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     9421 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     9481 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat
     9541 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     9601 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     9661 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     9721 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     9781 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     9841 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     9901 gagagtgaga ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg
     9961 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
    10021 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
    10081 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    10141 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
    10201 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
    10261 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
    10321 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
    10381 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
    10441 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
    10501 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
    10561 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
    10621 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
    10681 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
    10741 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    10801 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    10861 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    10921 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    10981 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    11041 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    11101 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    11161 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    11221 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    11281 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
    11341 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    11401 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    11461 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    11521 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
    11581 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
    11641 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
    11701 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
    11761 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
    11821 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
    11881 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
    11941 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
    12001 atgcagcaag ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
    12061 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
    12121 aagctccccg tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
    12181 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
    12241 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
    12301 tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    12361 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    12421 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta
    12481 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac
    12541 tacaaactta gtagt
//
