LOCUS       sequence-429717-       14820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector for dCas9-BFP-KRAB on Ef1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14820)
  TITLE     Plasmids for dCas9-Krab repression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14820)
  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            "Unpublished"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: pHR Ef1a-dCas9-BFP-KRAB
FEATURES             Location/Qualifiers
     source          1..14820
                     /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    2050..2167
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2222..3399
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2453..3391
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3348..3368
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3449..3458
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3455..3481,3482..3484,3485..5971,5972..5974,
                     5975..7558)
                     /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             3455..7558
                     /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             7562..7588
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7568..7594
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             7607..7627
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7634..7654
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7694..8389
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             8435..8620
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     misc_feature    8684..9272
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8737..8757)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9155..9166)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9275..9291)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9276..9292)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9382..9615
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(9640..9658)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(9641..9658)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(9948..9967)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(10181..10200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10300..10322
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10360..10378)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10446..10550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10551..10619,10620..11411)
                     /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     complement(10769..10788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11582..12170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12071..12090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12324..12341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(12411..12431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12416..12745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12596..12731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          13879..13944
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             14074..14094
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    14519..14653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14556..14575)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14610..14629
                     /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 tatcgccaaa tggcagtatt
     2041 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2161 aaattttcgg gtttattaca gggacagcag agatccagtt tggatcgata agcttgatat
     2221 cgaattctgt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     2281 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     2341 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     2401 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     2461 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     2521 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     2581 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     2641 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     2701 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     2761 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     2821 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     2881 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     2941 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     3001 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     3061 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     3121 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     3181 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     3241 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     3301 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     3361 agtggttcaa agtttttttc ttccatttca ggtgtcgtgg aattctcacg cgtctgcagg
     3421 atatcaagct tgcggtaccg cgggcccggc caccatggac aagaagtaca gcatcggcct
     3481 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     3541 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     3601 cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga gaaccgccag
     3661 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     3721 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     3781 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     3841 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     3901 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     3961 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     4021 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     4081 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     4141 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
     4201 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     4261 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     4321 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     4381 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     4441 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     4501 gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg gagccagcca
     4561 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     4621 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     4681 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     4741 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     4801 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     4861 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg ccagcgccca
     4921 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     4981 caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca aagtgaaata
     5041 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aagccatcgt
     5101 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     5161 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     5221 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     5281 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     5341 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     5401 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     5461 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     5521 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     5581 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     5641 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     5701 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     5761 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     5821 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     5881 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     5941 catcaaccgg ctgtccgact acgatgtgga cgctatcgtg cctcagagct ttctgaagga
     6001 cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca agagcgacaa
     6061 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc tgctgaatgc
     6121 caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag gcggcctgag
     6181 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     6241 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga acgacaaact
     6301 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     6361 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     6421 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     6481 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     6541 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     6601 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     6661 aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag tgctgtctat
     6721 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     6781 tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact gggaccctaa
     6841 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     6901 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     6961 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     7021 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     7081 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     7141 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     7201 cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc tggacgagat
     7261 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     7321 ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg ccgagaatat
     7381 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     7441 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     7501 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacgc
     7561 ctatccctat gacgtgcccg attatgccag cctgggcagc ggctccccca agaaaaaacg
     7621 caaggtggaa gatcctaaga aaaagcggaa agtggacggc attggtagtg ggagcaacgg
     7681 cagcagcgga tccagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg
     7741 caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
     7801 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
     7861 cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc
     7921 cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
     7981 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     8041 caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac
     8101 actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag
     8161 aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac
     8221 atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta
     8281 cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc
     8341 agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatg gcggtggcgg
     8401 agggatggat gctaagtcac taactgcctg gtcccggaca ctggtgacct tcaaggatgt
     8461 atttgtggac ttcaccaggg aggagtggaa gctgctggac actgctcagc agatcgtgta
     8521 cagaaatgtg atgctggaga actataagaa cctggtttcc ttgggttatc agcttactaa
     8581 gccagatgtg atcctccggt tggagaaggg agaagagccc tagtaaggcc gcgactctag
     8641 agtcgacctg caggcatgca agcttgatat caagcttatc gataatcaac ctctggatta
     8701 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     8761 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     8821 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     8881 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     8941 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     9001 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     9061 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     9121 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     9181 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     9241 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gagggaatta
     9301 attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     9361 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     9421 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     9481 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     9541 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     9601 ggaaaatctc tagcagcatc tagaattaat tccgtgtatt ctatagtgtc acctaaatcg
     9661 tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg acaatatgta
     9721 caagcctaat tgtgtagcat ctggcttact gaagcagacc ctatcatctc tctcgtaaac
     9781 tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt tctggtaacg ccgtcccgca
     9841 cccggaaatg gtcagcgaac caatcagcag ggtcatcgct agccagatcc tctacgccgg
     9901 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     9961 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
    10021 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
    10081 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
    10141 ggagtcgcat aagggagagc gtcgaatggt gcactctcag tacaatctgc tctgatgccg
    10201 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
    10261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
    10321 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
    10381 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
    10441 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
    10501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
    10561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
    10621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
    10681 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
    10741 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
    10801 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
    10861 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
    10921 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
    10981 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
    11041 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    11101 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    11161 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    11221 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    11281 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    11341 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    11401 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    11461 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    11521 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    11581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    11641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    11701 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    11761 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    11821 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    11881 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    11941 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    12001 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    12061 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    12121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    12181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    12241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    12301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
    12361 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg
    12421 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
    12481 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
    12541 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
    12601 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
    12661 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
    12721 ttttggaggc ctaggctttt gcaaaaagct tggacacaag acaggcttgc gagatatgtt
    12781 tgagaatacc actttatccc gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat
    12841 gtgaaatact ggtttttagt gcgccagatc tctataatct cgcgcaacct attttcccct
    12901 cgaacacttt ttaagccgta gataaacagg ctgggacact tcacatgagc gaaaaataca
    12961 tcgtcacctg ggacatgttg cagatccatg cacgtaaact cgcaagccga ctgatgcctt
    13021 ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact
    13081 ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt atttccagct acgatcacga
    13141 caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt
    13201 tattgatgac ctggtggata ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa
    13261 agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt
    13321 tgttgatatc ccgcaagata cctggattga acagccgtgg gatatgggcg tcgtattcgt
    13381 cccgccaatc tccggtcgct aatcttttca acgcctggca ctgccgggcg ttgttctttt
    13441 taacttcagg cgggttacaa tagtttccag taagtattct ggaggctgca tccatgacac
    13501 aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa acatcctgaa acctcgacgc
    13561 tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa
    13621 ccatccctca ctggtatcgc atgattaacc gtctgatgtg gatctggcgc ggcattgacc
    13681 cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg
    13741 atgatttata cgatacggtg attggctacc gtggcggcaa ctggatttat gagtgggccc
    13801 cggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca
    13861 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg
    13921 attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg
    13981 tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat
    14041 gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac
    14101 cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga
    14161 actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa
    14221 attatggaaa aatattctgt aacctttata agtaggcata acagttataa tcataacata
    14281 ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa
    14341 ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt
    14401 gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
    14461 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
    14521 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
    14581 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
    14641 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
    14701 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
    14761 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
