LOCUS       sequence-211595-       13936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector containing SFFV promoter, dCas9 that is 
            fused to 2x NLS and tagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13936)
  AUTHORS   Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, 
            Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, 
            Weissman JS, Qi LS
  TITLE     CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in 
            Eukaryotes.
  JOURNAL   Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 
            10.1016/j.cell.2013.06.044.
  PUBMED    23849981
REFERENCE   2  (bases 1 to 13936)
  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: "Cell. 2013 
            Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044."
COMMENT     Sequence Label: pHR-SFFV-dCas9-BFP
FEATURES             Location/Qualifiers
     source          1..13936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    678..803
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1300..1533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1718..1762
                     /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    2047..2164
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2317..2724
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      2793..2802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2799..2825,2826..2828,2829..5315,5316..5318,
                     5319..6902)
                     /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             2799..6902
                     /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             6906..6932
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6912..6938
                     /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             6951..6971
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6978..6998
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7038..7736
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    7797..8385
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7850..7870)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8268..8279)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8388..8404)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8389..8405)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8495..8728
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(8753..8771)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8754..8771)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(9061..9080)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(9294..9313)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9413..9435
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9473..9491)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9559..9663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9664..9732,9733..10524)
                     /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(9882..9901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10695..11283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11184..11203
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11437..11454
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(11524..11544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11529..11858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11709..11844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          12992..13057
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             13187..13207
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    13632..13766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13669..13688)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13723..13742
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(13934..13936,1..631)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
       61 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
      121 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
      181 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
      241 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
      301 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
      361 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
      421 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
      481 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      541 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      601 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
      661 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
      721 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
      781 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
      841 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
      901 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
      961 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     1021 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     1081 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     1141 cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     1981 agacagatcc attcgattag tgaacggatc tcgacggtat cgccaaatgg cagtattcat
     2041 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2101 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2161 ttttcgggtt tattacaggg acagcagaga tccagtttgg atcgataagc ttgatatcga
     2221 attcctgcag ccccgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     2281 aaagacccca cctgtaggtt tggcaagcta gctgcagtaa cgccattttg caaggcatgg
     2341 aaaaatacca aaccaagaat agagaagttc agatcaaggg cgggtacatg aaaatagcta
     2401 acgttgggcc aaacaggata tctgcggtga gcagtttcgg ccccggcccg gggccaagaa
     2461 cagatggtca ccgcagtttc ggccccggcc cgaggccaag aacagatggt ccccagatat
     2521 ggcccaaccc tcagcagttt cttaagaccc atcagatgtt tccaggctcc cccaaggacc
     2581 tgaaatgacc ctgcgcctta tttgaattaa ccaatcagcc tgcttctcgc ttctgttcgc
     2641 gcgcttctgc ttcccgagct ctataaaaga gctcacaacc cctcactcgg cgcgccagtc
     2701 ctccgacaga ctgagtcgcc cgggggggat ctggagctct cgagaattct cacgcgtctg
     2761 caggatatca agcttgcggt accgcgggcc cggccaccat ggacaagaag tacagcatcg
     2821 gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     2881 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     2941 tcggcgccct gctgttcgac agcggagaaa cagccgaggc cacccggctg aagagaaccg
     3001 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     3061 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     3121 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     3181 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     3241 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     3301 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     3361 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     3421 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     3481 agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc
     3541 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     3601 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     3661 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     3721 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     3781 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     3841 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacatcgat ggcggagcca
     3901 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     3961 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     4021 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     4081 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     4141 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     4201 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgccagcg
     4261 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     4321 tgcccaagca cagcctgctg tacgagtact tcaccgtgta caacgagctg accaaagtga
     4381 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaagcca
     4441 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     4501 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     4561 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     4621 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     4681 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     4741 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     4801 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     4861 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     4921 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     4981 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     5041 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     5101 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     5161 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     5221 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     5281 tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag agctttctga
     5341 aggacgactc catcgataac aaagtgctga ctcggagcga caagaaccgg ggcaagagcg
     5401 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgc cagctgctga
     5461 atgccaagct gattacccag aggaagttcg acaatctgac caaggccgag agaggcggcc
     5521 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     5581 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaacgaca
     5641 aactgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     5701 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     5761 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     5821 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     5881 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     5941 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     6001 aaacaggcga gatcgtgtgg gataagggcc gggactttgc caccgtgcgg aaagtgctgt
     6061 ctatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     6121 agtctatcct gcccaagagg aacagcgaca agctgatcgc cagaaagaag gactgggacc
     6181 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
     6241 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     6301 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     6361 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     6421 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
     6481 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     6541 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaaacac tacctggacg
     6601 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     6661 acaaggtgct gagcgcctac aacaagcaca gagacaagcc tatcagagag caggccgaga
     6721 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     6781 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     6841 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
     6901 acgcctatcc ctatgacgtg cccgattatg ccagcctggg cagcggctcc cccaagaaaa
     6961 aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt agtgggagca
     7021 acggcagcag cggatccagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
     7081 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
     7141 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     7201 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
     7261 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     7321 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     7381 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     7441 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     7501 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga
     7561 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     7621 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     7681 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt aattagtaag
     7741 gccgcgactc tagagtcgac ctgcaggcat gcaagcttga tatcaagctt atcgataatc
     7801 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     7861 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     7921 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     7981 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     8041 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     8101 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     8161 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     8221 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     8281 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     8341 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     8401 ctcgagggaa ttaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta
     8461 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     8521 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     8581 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     8641 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     8701 ttttagtcag tgtggaaaat ctctagcagc atctagaatt aattccgtgt attctatagt
     8761 gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta
     8821 acgacaatat gtacaagcct aattgtgtag catctggctt actgaagcag accctatcat
     8881 ctctctcgta aactgccgtc agagtcggtt tggttggacg aaccttctga gtttctggta
     8941 acgccgtccc gcacccggaa atggtcagcg aaccaatcag cagggtcatc gctagccaga
     9001 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     9061 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     9121 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     9181 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     9241 gcttcctaat gcaggagtcg cataagggag agcgtcgaat ggtgcactct cagtacaatc
     9301 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     9361 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     9421 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     9481 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     9541 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     9601 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     9661 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     9721 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     9781 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     9841 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     9901 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     9961 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
    10021 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
    10081 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
    10141 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
    10201 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
    10261 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
    10321 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
    10381 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
    10441 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
    10501 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
    10561 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
    10621 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
    10681 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
    10741 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
    10801 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
    10861 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
    10921 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
    10981 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
    11041 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
    11101 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
    11161 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
    11221 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
    11281 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
    11341 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
    11401 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
    11461 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
    11521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
    11581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
    11641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
    11701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
    11761 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
    11821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttggacac aagacaggct
    11881 tgcgagatat gtttgagaat accactttat cccgcgtcag ggagaggcag tgcgtaaaaa
    11941 gacgcggact catgtgaaat actggttttt agtgcgccag atctctataa tctcgcgcaa
    12001 cctattttcc cctcgaacac tttttaagcc gtagataaac aggctgggac acttcacatg
    12061 agcgaaaaat acatcgtcac ctgggacatg ttgcagatcc atgcacgtaa actcgcaagc
    12121 cgactgatgc cttctgaaca atggaaaggc attattgccg taagccgtgg cggtctgtac
    12181 cgggtgcgtt actggcgcgt gaactgggta ttcgtcatgt cgataccgtt tgtatttcca
    12241 gctacgatca cgacaaccag cgcgagctta aagtgctgaa acgcgcagaa ggcgatggcg
    12301 aaggcttcat cgttattgat gacctggtgg ataccggtgg tactgcggtt gcgattcgtg
    12361 aaatgtatcc aaaagcgcac tttgtcacca tcttcgcaaa accggctggt cgtccgctgg
    12421 ttgatgacta tgttgttgat atcccgcaag atacctggat tgaacagccg tgggatatgg
    12481 gcgtcgtatt cgtcccgcca atctccggtc gctaatcttt tcaacgcctg gcactgccgg
    12541 gcgttgttct ttttaacttc aggcgggtta caatagtttc cagtaagtat tctggaggct
    12601 gcatccatga cacaggcaaa cctgagcgaa accctgttca aaccccgctt taaacatcct
    12661 gaaacctcga cgctagtccg ccgctttaat cacggcgcac aaccgcctgt gcagtcggcc
    12721 cttgatggta aaaccatccc tcactggtat cgcatgatta accgtctgat gtggatctgg
    12781 cgcggcattg acccacgcga aatcctcgac gtccaggcac gtattgtgat gagcgatgcc
    12841 gaacgtaccg acgatgattt atacgatacg gtgattggct accgtggcgg caactggatt
    12901 tatgagtggg ccccggatct ttgtgaagga accttacttc tgtggtgtga cataattgga
    12961 caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg
    13021 tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga
    13081 atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc
    13141 atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag
    13201 aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt
    13261 gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact
    13321 gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta
    13381 taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa
    13441 ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata
    13501 tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt
    13561 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
    13621 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    13681 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    13741 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
    13801 aacttcccac cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa
    13861 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
    13921 caaacttagt agttgg
//
