LOCUS       sequence-379928-       14340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCas9-p300 with 2A Blast 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14340)
  AUTHORS   Omachi K, Miner JH
  TITLE     Comparative analysis of dCas9-VP64 variants and multiplexed guide 
            RNAs mediating CRISPR activation.
  JOURNAL   PLoS One. 2022 Jun 28;17(6):e0270008. doi: 
            10.1371/journal.pone.0270008. eCollection 2022.
  PUBMED    35763517
REFERENCE   2  (bases 1 to 14340)
  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: "PLoS One.";
            date: "2022-06-28"; volume: "17(6):e0270008. doi"; pages: " 
            10.1371/journal.pone.0270008. eCollection 2022"
COMMENT     Sequence Label: Lenti-EF1alpha-dCas9-p300_Blast
FEATURES             Location/Qualifiers
     source          1..14340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2440..2651
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2670..2679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2676..2702,2703..2705,2706..5192,5193..5195,
                     5196..6779)
                     /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             2676..6779
                     /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             6780..6827
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6834..8684
                     /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      8448..8457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8691..8714
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8724..8780
                     /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             8781..9179
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9195..9783
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9248..9268)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9666..9677)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9855..10088
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10114..10131)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10120..10344
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10390..10818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10477..10496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10687..10708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10827..10847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10832..11161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11012..11147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11074..11093
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11209..11256
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11275..11649
                     /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    11779..11900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11816..11835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11870..11889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11949..11965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11949..11965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11962..11984)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11973..11989
                     /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(11997..12003,12004..12021,12022..12027))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12042..12063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12180..12197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12351..12939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12431..12450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13110..13901,13902..13970))
                     /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     13733..13752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13971..14075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14150..14169)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gaggcgatag ggttaaggga gctagctagg
     2401 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     2461 cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc
     2521 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2581 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2641 ccagaacaca ggaccggttc tagagcgctg ccaccatgga caagaagtac agcatcggcc
     2701 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     2761 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     2821 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     2881 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     2941 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     3001 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     3061 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     3121 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     3181 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     3241 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     3301 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     3361 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     3421 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     3481 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     3541 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     3601 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     3661 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     3721 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     3781 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     3841 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     3901 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     3961 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     4021 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     4081 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     4141 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     4201 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     4261 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     4321 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     4381 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     4441 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     4501 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     4561 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     4621 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     4681 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     4741 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     4801 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     4861 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     4921 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     4981 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     5041 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     5101 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     5161 acatcaaccg gctgtccgac tacgatgtgg acgccatcgt gcctcagagc tttctgaagg
     5221 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     5281 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     5341 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     5401 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     5461 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     5521 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     5581 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     5641 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     5701 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     5761 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     5821 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     5881 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     5941 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     6001 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     6061 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     6121 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     6181 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     6241 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     6301 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     6361 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     6421 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     6481 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     6541 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     6601 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     6661 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     6721 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     6781 agcgacctgc cgccacaaag aaggctggac aggctaagaa gaagaaagga tccattttca
     6841 aaccagaaga actacgacag gcactgatgc caactttgga ggcactttac cgtcaggatc
     6901 cagaatccct tccctttcgt caacctgtgg accctcagct tttaggaatc cctgattact
     6961 ttgatattgt gaagagcccc atggatcttt ctaccattaa gaggaagtta gacactggac
     7021 agtatcagga gccctggcag tatgtcgatg atatttggct tatgttcaat aatgcctggt
     7081 tatataaccg gaaaacatca cgggtataca aatactgctc caagctctct gaggtctttg
     7141 aacaagaaat tgacccagtg atgcaaagcc ttggatactg ttgtggcaga aagttggagt
     7201 tctctccaca gacactgtgt tgctacggca aacagttgtg cacaatacct cgtgatgcca
     7261 cttattacag ttaccagaac aggtatcatt tctgtgagaa gtgtttcaat gagatccaag
     7321 gggagagcgt ttctttgggg gatgaccctt cccagcctca aactacaata aataaagaac
     7381 aattttccaa gagaaaaaat gacacactgg atcctgaact gtttgttgaa tgtacagagt
     7441 gcggaagaaa gatgcatcag atctgtgtcc ttcaccatga gatcatctgg cctgctggat
     7501 tcgtctgtga tggctgttta aagaaaagtg cacgaactag gaaagaaaat aagttttctg
     7561 ctaaaaggtt gccatctacc agacttggca cctttctaga gaatcgtgtg aatgactttc
     7621 tgaggcgaca gaatcaccct gagtcaggag aggtcactgt tagagtagtt catgcttctg
     7681 acaaaaccgt ggaagtaaaa ccaggcatga aagcaaggtt tgtggacagt ggagagatgg
     7741 cagaatcctt tccataccga accaaagccc tctttgcctt tgaagaaatt gatggtgttg
     7801 acctgtgctt ctttggcatg catgttcaag agtatggctc tgactgccct ccacccaacc
     7861 agaggagagt atacatatct tacctcgata gtgttcattt cttccgtcct aaatgcttga
     7921 ggactgcagt ctatcatgaa atcctaattg gatatttaga atatgtcaag aaattaggtt
     7981 acacaacagg gcatatttgg gcatgtccac caagtgaggg agatgattat atcttccatt
     8041 gccatcctcc tgaccagaag atacccaagc ccaagcgact gcaggaatgg tacaaaaaaa
     8101 tgcttgacaa ggctgtatca gagcgtattg tccatgacta caaggatatt tttaaacaag
     8161 ctactgaaga tagattaaca agtgcaaagg aattgcctta tttcgagggt gatttctggc
     8221 ccaatgttct ggaagaaagc attaaggaac tggaacagga ggaagaagag agaaaacgag
     8281 aggaaaacac cagcaatgaa agcacagatg tgaccaaggg agacagcaaa aatgctaaaa
     8341 agaagaataa taagaaaacc agcaaaaata agagcagcct gagtaggggc aacaagaaga
     8401 aacccgggat gcccaatgta tctaacgacc tctcacagaa actatatgcc accatggaga
     8461 agcataaaga ggtcttcttt gtgatccgcc tcattgctgg ccctgctgcc aactccctgc
     8521 ctcccattgt tgatcctgat cctctcatcc cctgcgatct gatggatggt cgggatgcgt
     8581 ttctcacgct ggcaagggac aagcacctgg agttctcttc actccgaaga gcccagtggt
     8641 ccaccatgtg catgctggtg gagctgcaca cgcagagcca ggacggatcc gattacaaag
     8701 acgatgacga taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     8761 tcgaagagaa tcctggaccg atggccaagc ctttgtctca agaagaatcc accctcattg
     8821 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     8881 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     8941 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     9001 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     9061 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     9121 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaaa
     9181 cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     9241 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     9301 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     9361 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     9421 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     9481 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     9541 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     9601 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     9661 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     9721 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     9781 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     9841 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     9901 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     9961 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
    10021 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
    10081 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
    10141 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
    10201 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
    10261 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
    10321 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
    10381 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
    10441 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
    10501 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
    10561 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    10621 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    10681 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    10741 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    10801 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
    10861 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
    10921 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
    10981 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
    11041 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    11101 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
    11161 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
    11221 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
    11281 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
    11341 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
    11401 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
    11461 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
    11521 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
    11581 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
    11641 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
    11701 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
    11761 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    11821 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    11881 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
    11941 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    12001 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    12061 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    12121 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    12181 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    12241 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    12301 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    12361 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    12421 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    12481 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    12541 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    12601 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    12661 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    12721 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    12781 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    12841 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    12901 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    12961 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    13021 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    13081 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    13141 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    13201 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    13261 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    13321 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    13381 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    13441 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    13501 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    13561 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    13621 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    13681 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    13741 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    13801 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    13861 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    13921 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    13981 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    14041 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    14101 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
    14161 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
    14221 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
    14281 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
