LOCUS       sequence-322468-       14383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9-VP64 activator with 2A GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14383)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 14383)
  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: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: dCAS9-VP64_GFP
FEATURES             Location/Qualifiers
     source          1..14383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2682..3860
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2913..3851
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3808..3828
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3879..3899
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3909..3932,3933..3935,3936..6422,6423..6425,
                     6426..8009)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3909..8009
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8064..8084
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8109..8258
                     /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             8280..8333
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8334..8336,8337..9050)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8334..8336,8337..9047)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8637..8656)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8984..9005
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    9075..9663
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9128..9148)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9546..9557)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9666..9682)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9667..9683)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10188..10368
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10394..10411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10400..10624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10670..11098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10757..10776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10967..10988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11107..11127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11112..11441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11292..11427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11354..11373
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11489..11536
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11555..11929
                     /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    12059..12180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12096..12115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12150..12169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12229..12245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12229..12245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12242..12264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12253..12269
                     /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(12277..12283,12284..12301,12302..12307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12322..12343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12460..12477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12631..13219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12711..12730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13390..14181,14182..14250))
                     /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     14013..14032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14251..14355)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt
     2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
     2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     3001 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     3061 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     3121 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     3181 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     3241 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     3301 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     3361 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     3421 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3481 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3541 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3601 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3661 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3721 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3781 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3841 ttccatttca ggtgtcgtga cgtacggcca ccatgagccc caagaagaag agaaaggtgg
     3901 aggccagcga caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg
     3961 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     4021 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     4081 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     4141 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     4201 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     4261 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     4321 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     4381 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     4441 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     4501 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     4561 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     4621 gcaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     4681 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     4741 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     4801 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     4861 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     4921 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     4981 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     5041 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     5101 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     5161 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     5221 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     5281 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     5341 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     5401 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     5461 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     5521 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     5581 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     5641 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     5701 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     5761 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     5821 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     5881 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     5941 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     6001 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     6061 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     6121 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     6181 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     6241 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     6301 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     6361 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     6421 acgctatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     6481 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     6541 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     6601 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     6661 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     6721 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     6781 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     6841 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     6901 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     6961 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     7021 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     7081 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     7141 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     7201 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     7261 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     7321 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     7381 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     7441 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     7501 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     7561 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     7621 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     7681 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     7741 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     7801 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     7861 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     7921 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     7981 ggatcgacct gtctcagctg ggaggcgaca gcgctggagg aggtggaagc ggaggaggag
     8041 gaagcggagg aggaggtagc ggacctaaga aaaagaggaa ggtggcggcc gctggatccg
     8101 gacgggctga cgcattggac gattttgatc tggatatgct gggaagtgac gccctcgatg
     8161 attttgacct tgacatgctt ggttcggatg cccttgatga ctttgacctc gacatgctcg
     8221 gcagtgacgc ccttgatgat ttcgacctgg acatgctgat taacgctagc ggcagtggag
     8281 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccagtgagca
     8341 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     8401 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     8461 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     8521 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     8581 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     8641 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     8701 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     8761 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     8821 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     8881 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     8941 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     9001 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gaattcgata
     9061 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     9121 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     9181 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     9241 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     9301 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     9361 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     9421 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     9481 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     9541 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     9601 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     9661 cgcatcgata ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata
     9721 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     9781 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     9841 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     9901 atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca
     9961 caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag
    10021 ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg
    10081 tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc
    10141 gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt
    10201 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
    10261 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
    10321 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
    10381 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
    10441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
    10501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
    10561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
    10621 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
    10681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
    10741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
    10801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
    10861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    10921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
    10981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
    11041 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
    11101 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
    11161 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
    11221 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
    11281 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
    11341 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
    11401 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
    11461 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
    11521 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
    11581 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
    11641 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
    11701 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
    11761 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
    11821 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
    11881 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
    11941 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
    12001 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
    12061 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
    12121 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
    12181 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
    12241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    12301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    12361 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    12421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    12481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    12541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    12601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    12661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    12721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    12781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    12841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    12901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    12961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
    13021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
    13081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    13141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    13201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    13261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
    13321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    13381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    13441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    13501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    13561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    13621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    13681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    13741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    13801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    13861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    13921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    13981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    14041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    14101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    14161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    14221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    14281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    14341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
