LOCUS       sequence-322487-       15707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2nd Generation Lentiviral vector. Expresses an N-terminal p300-dCas9
            fusion protein and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15707)
  AUTHORS   Li K, Liu Y, Cao H, Zhang Y, Gu Z, Liu X, Yu A, Kaphle P, Dickerson 
            KE, Ni M, Xu J
  TITLE     Interrogation of enhancer function by enhancer-targeting CRISPR 
            epigenetic editing.
  JOURNAL   Nat Commun. 2020 Jan 24;11(1):485. doi: 10.1038/s41467-020-14362-5.
  PUBMED    31980609
REFERENCE   2  (bases 1 to 15707)
  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: "Nat 
            Commun."; date: "2020-01-24"; volume: "11(1):485. doi"; pages: " 
            10.1038/s41467-020-14362-5"
COMMENT     Sequence Label: pHR_TRE3G-p300-dCas9-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..15707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2050..2167
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2229..2578
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2237..2255
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2273..2291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2309..2327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2345..2363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2381..2399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2417..2435
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2453..2471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2517..2541
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2635..4485
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      4249..4258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4507..4533,4534..4536,4537..7023,7024..7026,
                     7027..8610)
                     /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             4507..8610
                     /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             8614..8640
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8620..8646
                     /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             8659..8679
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8686..8706
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8746..8802
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8803..9513
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8948..8966)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9216..9235
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    9571..10159
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9624..9644)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10042..10053)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10162..10178)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10163..10179)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10269..10502
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(10527..10545)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(10528..10545)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10835..10854)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(11068..11087)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11187..11209
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11247..11265)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11333..11437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11438..11506,11507..12298)
                     /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(11656..11675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12469..13057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12958..12977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13211..13228
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(13298..13318)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13303..13632
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13483..13618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          14766..14831
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             14961..14981
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    15406..15540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(15443..15462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     15497..15516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccaaa tggcagtatt
     2041 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2161 aaattttcgg gtttattaca gggacagcag agatccagtt tatcgatgag gccctttcgt
     2221 cttcactcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     2281 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     2341 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     2401 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     2461 agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct
     2521 cgtttagtga accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttac
     2581 gcgtctgcag gatatcaagc ttgcggtacc gcgggcccgg gatccgccac catgattttc
     2641 aaaccagaag aactacgaca ggcactgatg ccaactttgg aggcacttta ccgtcaggat
     2701 ccagaatccc ttccctttcg tcaacctgtg gaccctcagc ttttaggaat ccctgattac
     2761 tttgatattg tgaagagccc catggatctt tctaccatta agaggaagtt agacactgga
     2821 cagtatcagg agccctggca gtatgtcgat gatatttggc ttatgttcaa taatgcctgg
     2881 ttatataacc ggaaaacatc acgggtatac aaatactgct ccaagctctc tgaggtcttt
     2941 gaacaagaaa ttgacccagt gatgcaaagc cttggatact gttgtggcag aaagttggag
     3001 ttctctccac agacactgtg ttgctacggc aaacagttgt gcacaatacc tcgtgatgcc
     3061 acttattaca gttaccagaa caggtatcat ttctgtgaga agtgtttcaa tgagatccaa
     3121 ggggagagcg tttctttggg ggatgaccct tcccagcctc aaactacaat aaataaagaa
     3181 caattttcca agagaaaaaa tgacacactg gatcctgaac tgtttgttga atgtacagag
     3241 tgcggaagaa agatgcatca gatctgtgtc cttcaccatg agatcatctg gcctgctgga
     3301 ttcgtctgtg atggctgttt aaagaaaagt gcacgaacta ggaaagaaaa taagttttct
     3361 gctaaaaggt tgccatctac cagacttggc acctttctag agaatcgtgt gaatgacttt
     3421 ctgaggcgac agaatcaccc tgagtcagga gaggtcactg ttagagtagt tcatgcttct
     3481 gacaaaaccg tggaagtaaa accaggcatg aaagcaaggt ttgtggacag tggagagatg
     3541 gcagaatcct ttccataccg aaccaaagcc ctctttgcct ttgaagaaat tgatggtgtt
     3601 gacctgtgct tctttggcat gcatgttcaa gagtatggct ctgactgccc tccacccaac
     3661 cagaggagag tatacatatc ttacctcgat agtgttcatt tcttccgtcc taaatgcttg
     3721 aggactgcag tctatcatga aatcctaatt ggatatttag aatatgtcaa gaaattaggt
     3781 tacacaacag ggcatatttg ggcatgtcca ccaagtgagg gagatgatta tatcttccat
     3841 tgccatcctc ctgaccagaa gatacccaag cccaagcgac tgcaggaatg gtacaaaaaa
     3901 atgcttgaca aggctgtatc agagcgtatt gtccatgact acaaggatat ttttaaacaa
     3961 gctactgaag atagattaac aagtgcaaag gaattgcctt atttcgaggg tgatttctgg
     4021 cccaatgttc tggaagaaag cattaaggaa ctggaacagg aggaagaaga gagaaaacga
     4081 gaggaaaaca ccagcaatga aagcacagat gtgaccaagg gagacagcaa aaatgctaaa
     4141 aagaagaata ataagaaaac cagcaaaaat aagagcagcc tgagtagggg caacaagaag
     4201 aaacccggga tgcccaatgt atctaacgac ctctcacaga aactatatgc caccatggag
     4261 aagcataaag aggtcttctt tgtgatccgc ctcattgctg gccctgctgc caactccctg
     4321 cctcccattg ttgatcctga tcctctcatc ccctgcgatc tgatggatgg tcgggatgcg
     4381 tttctcacgc tggcaaggga caagcacctg gagttctctt cactccgaag agcccagtgg
     4441 tccaccatgt gcatgctggt ggagctgcac acgcagagcc aggacacgcg ttccggcgga
     4501 gggtcgatgg acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg
     4561 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     4621 gaccggcaca gcatcaagaa gaacctgatc ggcgccctgc tgttcgacag cggagaaaca
     4681 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     4741 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     4801 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     4861 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     4921 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     4981 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     5041 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     5101 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     5161 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     5221 ggcaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     5281 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     5341 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     5401 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     5461 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     5521 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     5581 tacgccggct acatcgatgg cggagccagc caggaagagt tctacaagtt catcaagccc
     5641 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     5701 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     5761 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     5821 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     5881 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     5941 gaggaagtgg tggacaaggg cgccagcgcc cagagcttca tcgagcggat gaccaacttc
     6001 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     6061 accgtgtaca acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     6121 ttcctgagcg gcgagcagaa aaaagccatc gtggacctgc tgttcaagac caaccggaaa
     6181 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     6241 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     6301 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     6361 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     6421 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     6481 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     6541 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     6601 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     6661 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     6721 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     6781 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     6841 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     6901 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     6961 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     7021 gacgctatcg tgcctcagag ctttctgaag gacgactcca tcgataacaa agtgctgact
     7081 cggagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     7141 atgaagaact actggcgcca gctgctgaat gccaagctga ttacccagag gaagttcgac
     7201 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     7261 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     7321 atgaacacta agtacgacga gaacgacaaa ctgatccggg aagtgaaagt gatcaccctg
     7381 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     7441 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     7501 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     7561 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     7621 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     7681 aagcggcctc tgatcgagac aaacggcgaa acaggcgaga tcgtgtggga taagggccgg
     7741 gactttgcca ccgtgcggaa agtgctgtct atgccccaag tgaatatcgt gaaaaagacc
     7801 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgacaag
     7861 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     7921 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     7981 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     8041 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     8101 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     8161 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     8221 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     8281 gtggaacagc acaaacacta cctggacgag atcatcgagc agatcagcga gttctccaag
     8341 agagtgatcc tggccgacgc taatctggac aaggtgctga gcgcctacaa caagcacaga
     8401 gacaagccta tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     8461 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     8521 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     8581 cggatcgacc tgtctcagct gggaggcgac gcctatccct atgacgtgcc cgattatgcc
     8641 agcctgggca gcggctcccc caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg
     8701 aaagtggacg gcattggtag tgggagcaac ggcagcagcg gatccgctac taacttcagc
     8761 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatggtgag caagggcgag
     8821 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     8881 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     8941 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     9001 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     9061 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     9121 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     9181 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     9241 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     9301 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     9361 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     9421 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     9481 tccaccggcg gcatggacga gctgtacaag taaggccgcg actctagagt cgacctgcag
     9541 gcatgcaagc ttgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     9601 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     9661 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     9721 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     9781 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     9841 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     9901 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     9961 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
    10021 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
    10081 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
    10141 ctttgggccg cctccccgca tcgataccgt cgacctcgag ggaattaatt cgagctcggt
    10201 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    10261 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
    10321 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
    10381 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
    10441 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
    10501 cagcatctag aattaattcc gtgtattcta tagtgtcacc taaatcgtat gtgtatgata
    10561 cataaggtta tgtattaatt gtagccgcgt tctaacgaca atatgtacaa gcctaattgt
    10621 gtagcatctg gcttactgaa gcagacccta tcatctctct cgtaaactgc cgtcagagtc
    10681 ggtttggttg gacgaacctt ctgagtttct ggtaacgccg tcccgcaccc ggaaatggtc
    10741 agcgaaccaa tcagcagggt catcgctagc cagatcctct acgccggacg catcgtggcc
    10801 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
    10861 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
    10921 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
    10981 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
    11041 ggagagcgtc gaatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
    11101 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
    11161 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
    11221 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
    11281 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
    11341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
    11401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
    11461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
    11521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
    11581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
    11641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
    11701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
    11761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
    11821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
    11881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
    11941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
    12001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
    12061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
    12121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
    12181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
    12241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
    12301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    12361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    12421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    12481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    12541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    12601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    12661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    12721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    12781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    12841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    12901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    12961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    13021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    13081 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
    13141 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
    13201 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
    13261 tctccccgcg cgttggccga ttcattaatg cagctgtgga atgtgtgtca gttagggtgt
    13321 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
    13381 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    13441 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
    13501 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
    13561 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
    13621 ggcttttgca aaaagcttgg acacaagaca ggcttgcgag atatgtttga gaataccact
    13681 ttatcccgcg tcagggagag gcagtgcgta aaaagacgcg gactcatgtg aaatactggt
    13741 ttttagtgcg ccagatctct ataatctcgc gcaacctatt ttcccctcga acacttttta
    13801 agccgtagat aaacaggctg ggacacttca catgagcgaa aaatacatcg tcacctggga
    13861 catgttgcag atccatgcac gtaaactcgc aagccgactg atgccttctg aacaatggaa
    13921 aggcattatt gccgtaagcc gtggcggtct gtaccgggtg cgttactggc gcgtgaactg
    13981 ggtattcgtc atgtcgatac cgtttgtatt tccagctacg atcacgacaa ccagcgcgag
    14041 cttaaagtgc tgaaacgcgc agaaggcgat ggcgaaggct tcatcgttat tgatgacctg
    14101 gtggataccg gtggtactgc ggttgcgatt cgtgaaatgt atccaaaagc gcactttgtc
    14161 accatcttcg caaaaccggc tggtcgtccg ctggttgatg actatgttgt tgatatcccg
    14221 caagatacct ggattgaaca gccgtgggat atgggcgtcg tattcgtccc gccaatctcc
    14281 ggtcgctaat cttttcaacg cctggcactg ccgggcgttg ttctttttaa cttcaggcgg
    14341 gttacaatag tttccagtaa gtattctgga ggctgcatcc atgacacagg caaacctgag
    14401 cgaaaccctg ttcaaacccc gctttaaaca tcctgaaacc tcgacgctag tccgccgctt
    14461 taatcacggc gcacaaccgc ctgtgcagtc ggcccttgat ggtaaaacca tccctcactg
    14521 gtatcgcatg attaaccgtc tgatgtggat ctggcgcggc attgacccac gcgaaatcct
    14581 cgacgtccag gcacgtattg tgatgagcga tgccgaacgt accgacgatg atttatacga
    14641 tacggtgatt ggctaccgtg gcggcaactg gatttatgag tgggccccgg atctttgtga
    14701 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
    14761 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
    14821 gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
    14881 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
    14941 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
    15001 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
    15061 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
    15121 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta
    15181 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
    15241 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
    15301 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
    15361 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
    15421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    15481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
    15541 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
    15601 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
    15661 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
