LOCUS       sequence-101643-       13824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RNA-Guided, Nuclease-Inactive 
            VP64:dCas9-BFP:VP64—VdC9BV—Fusion Protein to Enable Transactivation 
            of Endogenous Genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13824)
  AUTHORS   Chakraborty S, Ji H, Kabadi AM, Gersbach CA, Christoforou N, Leong 
            KW
  TITLE     A CRISPR/Cas9-Based System for Reprogramming Cell Lineage 
            Specification.
  JOURNAL   Stem Cell Reports. 2014 Dec 9;3(6):940-7. doi: 
            10.1016/j.stemcr.2014.09.013. Epub 2014 Oct 23.
  PUBMED    25448066
REFERENCE   2  (bases 1 to 13824)
  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"; doi: 
            "10.1016/j.stemcr.2014.09.013"; journalName: "Stem Cell Reports"; 
            date: "2014-12-9- 9"; volume: "3"; issue: "6"; pages: "940-7"
COMMENT     Sequence Label: TetO-FUW-VdC9BV
FEATURES             Location/Qualifiers
     source          1..13824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    27..251
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      297..725
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(384..403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     594..615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(734..754)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        739..1068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      919..1054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     981..1000
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1116..1163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1182..1556
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1686..1807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1723..1742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1777..1796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1856..1872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1856..1872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1869..1891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1880..1896
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1904..1910,1911..1928,1929..1934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1949..1970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2087..2104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2258..2846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2338..2357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3017..3808,3809..3877))
                     /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     3640..3659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3878..3982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4057..4076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4248..4627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4629..4827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4782..4802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4845..5025
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5072..5197
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5690..5923
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6108..6152
                     /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    6450..6567
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(6621..6639,6640..6662,6663..6681,6682..6704,6705..6723,
                     6724..6746,6747..6765,6766..6788,6789..6807,6808..6830,
                     6831..6849,6850..6872,6873..6891)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        6924..6961
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6958..6982
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7064..7073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7088..7237
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             7244..7267
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(7313..7339,7340..7342,7343..9829,9830..9832,
                     9833..11416)
                     /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             11426..11446
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11456..12151
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             12161..12181
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12239..12259
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12269..12292
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12308..12457
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     misc_feature    12526..13114
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12579..12599)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12997..13008)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13117..13133)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13118..13134)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             13639..13819
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
       61 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
      121 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
      181 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
      241 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
      301 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      361 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      421 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      481 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      541 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      721 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     1021 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     1081 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     1141 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     1201 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     1261 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     1321 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     1381 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     1441 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     1501 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     1561 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     1621 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     1681 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1741 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1801 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     1861 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1921 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1981 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2041 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2101 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2161 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2221 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3961 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     4021 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     4081 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     4141 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     4201 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     4261 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     4321 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     4381 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     4441 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     4501 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     4561 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     4621 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     4681 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     4741 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     4801 tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag
     4861 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     4921 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     4981 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     5041 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     5101 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     5161 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     5221 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     5281 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     5341 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     5401 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     5461 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     5521 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     5581 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     5641 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     5701 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     5761 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     5821 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     5881 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     5941 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     6001 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     6061 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     6121 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     6181 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     6241 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     6301 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     6361 agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc
     6421 tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     6481 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     6541 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt
     6601 tggttactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     6661 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     6721 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     6781 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     6841 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     6901 gtcgagctcg gtacccgggt cgagtaggcg tgtacggtgg gaggcctata taagcagagc
     6961 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     7021 aagacaccgg gaccgatcca gcctccgcgg ccccgaattg ctagccacca tgggcagtgg
     7081 gagagccgac gcccttgacg atttcgatct cgatatgttg ggtagcgacg ccctcgacga
     7141 cttcgacctg gatatgctgg gctcagatgc attggatgac ttcgatctgg acatgctggg
     7201 atcagacgcg ctggacgact ttgacctgga tatgctcatc aacgactata aggacgatga
     7261 cgataaggga ggtggaggaa gcggcggtgg aggcagcggg ggcggtggct ctatggataa
     7321 gaaatactct atcggactcg ccattggcac aaactctgtt ggctgggctg tcataaccga
     7381 tgagtataag gtgccaagca agaaattcaa ggttctcggt aataccgacc gccactccat
     7441 caaaaagaat ctcatcggcg cccttctgtt tgacagcggc gagaccgcag aagctactcg
     7501 gctgaaacgg accgctcgaa gaagatacac acgacggaag aacaggatat gctacttgca
     7561 ggaaatcttt agtaacgaaa tggctaaagt cgatgatagt tttttccaca gactcgagga
     7621 aagtttcctg gtcgaggagg acaagaagca cgaacgacat ccgatcttcg ggaacatcgt
     7681 cgacgaggtg gcctatcacg agaaataccc aaccatctac catcttcgca aaaagctggt
     7741 ggactcaacc gacaaggcag acctccggct tatctacctg gccctggccc acatgatcaa
     7801 gttcagaggc cacttcctga tcgagggcga cctcaatcct gacaatagcg atgtggataa
     7861 actgttcatc cagctggtgc agacttacaa ccagctcttt gaagagaacc ccatcaatgc
     7921 aagcggagtc gatgccaagg ccattctgtc agcccggctg tcaaagagcc gcagacttga
     7981 gaatcttatc gctcagctgc cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc
     8041 tctttcactt gggctgactc ccaatttcaa gtctaatttc gacctggcag aggatgccaa
     8101 gctgcaactg tccaaggaca cctatgatga cgatctcgac aacctcctgg cccagatcgg
     8161 tgaccaatac gccgaccttt tccttgctgc taagaatctt tctgacgcca tcctgctgtc
     8221 tgacattctc cgcgtgaaca ctgaaatcac caaggcccct ctttcagctt caatgattaa
     8281 gcggtatgat gagcaccacc aggacctgac cctgcttaag gcactcgtcc ggcagcagct
     8341 tccggagaag tacaaggaaa tcttctttga ccagtcaaag aatggatacg ccggctacat
     8401 cgacggaggt gcctcccaag aggaatttta taagtttatc aaacctatcc ttgagaagat
     8461 ggacggcacc gaagagctcc tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg
     8521 cactttcgac aatgggagca ttccccacca gatccatctt ggggagcttc acgccatcct
     8581 tcggcgccaa gaggacttct acccctttct taaggacaac agggagaaga ttgagaaaat
     8641 tctcactttc cgcatcccct actacgtggg acccctcgcc agaggaaata gccggtttgc
     8701 ttggatgacc agaaagtcag aagaaactat cactccctgg aacttcgaag aggtggtgga
     8761 caagggagcc agcgctcagt cattcatcga acggatgact aacttcgata agaacctccc
     8821 caatgagaag gtcctgccga aacattccct gctctacgag tactttaccg tgtacaacga
     8881 gctgaccaag gtgaaatatg tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga
     8941 acaaaagaag gcaattgtgg accttctgtt caagaccaat agaaaggtga ccgtgaagca
     9001 gctgaaggag gactatttca agaaaattga atgcttcgac tctgtggaga ttagcggggt
     9061 cgaagatcgg ttcaacgcaa gcctgggtac ctaccatgat ctgcttaaga tcatcaagga
     9121 caaggatttt ctggacaatg aggagaacga ggacatcctt gaggacattg tcctgactct
     9181 cactctgttc gaggaccggg aaatgatcga ggagaggctt aagacctacg cccatctgtt
     9241 cgacgataaa gtgatgaagc aacttaaacg gagaagatat accggatggg gacgccttag
     9301 ccgcaaactc atcaacggaa tccgggacaa acagagcgga aagaccattc ttgatttcct
     9361 taagagcgac ggattcgcta atcgcaactt catgcaactt atccatgatg attccctgac
     9421 ctttaaggag gacatccaga aggcccaagt gtctggacaa ggtgactcac tgcacgagca
     9481 tatcgcaaat ctggctggtt cacccgctat taagaagggt attctccaga ccgtgaaagt
     9541 cgtggacgag ctggtcaagg tgatgggtcg ccataaacca gagaacattg tcatcgagat
     9601 ggccagggaa aaccagacta cccagaaggg acagaagaac agcagggagc ggatgaaaag
     9661 aattgaggaa gggattaagg agctcgggtc acagatcctt aaagagcacc cggtggaaaa
     9721 cacccagctt cagaatgaga agctctatct gtactacctt caaaatggac gcgatatgta
     9781 tgtggaccaa gagcttgata tcaacaggct ctcagactac gacgtggacg ccatcgtccc
     9841 tcagagcttc ctcaaagacg actcaattga caataaggtg ctgactcgct cagacaagaa
     9901 ccggggaaag tcagataacg tgccctcaga ggaagtcgtg aaaaagatga agaactattg
     9961 gcgccagctt ctgaacgcaa agctgatcac tcagcggaag ttcgacaatc tcactaaggc
    10021 tgagaggggc ggactgagcg aactggacaa agcaggattc attaaacggc aacttgtgga
    10081 gactcggcag attactaaac atgtcgccca aatccttgac tcacgcatga ataccaagta
    10141 cgacgaaaac gacaaactta tccgcgaggt gaaggtgatt accctgaagt ccaagctggt
    10201 cagcgatttc agaaaggact ttcaattcta caaagtgcgg gagatcaata actatcatca
    10261 tgctcatgac gcatatctga atgccgtggt gggaaccgcc ctgatcaaga agtacccaaa
    10321 gctggaaagc gagttcgtgt acggagacta caaggtctac gacgtgcgca agatgattgc
    10381 caaatctgag caggagatcg gaaaggccac cgcaaagtac ttcttctaca gcaacatcat
    10441 gaatttcttc aagaccgaaa tcacccttgc aaacggtgag atccggaaga ggccgctcat
    10501 cgagactaat ggggagactg gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt
    10561 gcgcaaagtg ctttctatgc ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg
    10621 aggcttttct aaggaatcaa tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa
    10681 gaaggattgg gaccctaaga agtacggcgg attcgattca ccaactgtgg cttattctgt
    10741 cctggtcgtg gctaaggtgg aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact
    10801 gctgggtatc accattatgg agcgcagctc cttcgagaag aacccaattg actttctcga
    10861 agccaaaggt tacaaggaag tcaagaagga ccttatcatc aagctcccaa agtatagcct
    10921 gttcgaactg gagaatgggc ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg
    10981 taatgagctg gctctcccct ccaagtacgt gaatttcctc taccttgcaa gccattacga
    11041 gaagctgaag gggagccccg aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa
    11101 gcattatctg gacgagatca ttgagcagat ttccgagttt tctaaacgcg tcattctcgc
    11161 tgatgccaac ctcgataaag tccttagcgc atacaataag cacagagaca aaccaattcg
    11221 ggagcaggct gagaatatca tccacctgtt caccctcacc aatcttggtg cccctgccgc
    11281 attcaagtac ttcgacacca ccatcgaccg gaaacgctat acctccacca aagaagtgct
    11341 ggacgccacc ctcatccacc agagcatcac cggactttac gaaactcgga ttgacctctc
    11401 acagctcgga ggggatgagg gagctcccaa gaaaaagcgc aaggtaggta gttccagcga
    11461 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca
    11521 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
    11581 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct
    11641 ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc
    11701 cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
    11761 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
    11821 ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt
    11881 caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa
    11941 gctcgtgggc gggagccatc tgatcgcaaa catcaagacc acatatagat ccaagaaacc
    12001 cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa
    12061 ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga
    12121 cctccctagc aaactggggc acaagcttaa tgagggagct ccaaagaaga agcgtaaggt
    12181 aggtagttcc ggtggaggag gttctggagg cggtggaagt ggtggcggag gtagcagccc
    12241 caagaagaag agaaaggtgg aggccagcga ctacaaggat gacgacgata aaggttccgg
    12301 acgggctgac gcattggacg attttgatct ggatatgctg ggaagtgacg ccctcgatga
    12361 ttttgacctt gacatgcttg gttcggatgc ccttgatgac tttgacctcg acatgctcgg
    12421 cagtgacgcc cttgatgatt tcgacctgga catgctgatt aactaactcg agaccaggat
    12481 gggcaccaaa ttcaggtctg acaattcgat atcaagctta tcgataatca acctctggat
    12541 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
    12601 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
    12661 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
    12721 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
    12781 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
    12841 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
    12901 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
    12961 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
    13021 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
    13081 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta
    13141 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
    13201 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
    13261 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
    13321 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
    13381 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg
    13441 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat
    13501 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag
    13561 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
    13621 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    13681 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    13741 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    13801 gtgtggaaaa tctctagcag ggcc
//
