LOCUS       sequence-388206-       14556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding VP64-dCas9-p300 with 2A Blast 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14556)
  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 14556)
  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-VP64-dCas9-p300_Blast
FEATURES             Location/Qualifiers
     source          1..14556
                     /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"
     CDS             2679..2699
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2724..2873
                     /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             join(2895..2918,2919..2921,2922..5408,5409..5411,
                     5412..6995)
                     /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             2895..6995
                     /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             6996..7043
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7050..8900
                     /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      8664..8673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8907..8930
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8940..8996
                     /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             8997..9395
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9411..9999
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9464..9484)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9882..9893)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10071..10304
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10330..10347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10336..10560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10606..11034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10693..10712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10903..10924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11043..11063)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11048..11377
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11228..11363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11290..11309
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11425..11472
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11491..11865
                     /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    11995..12116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12032..12051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12086..12105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12165..12181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12165..12181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12178..12200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12189..12205
                     /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(12213..12219,12220..12237,12238..12243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12258..12279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12396..12413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12567..13155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12647..12666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13326..14117,14118..14186))
                     /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     13949..13968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14187..14291)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14366..14385)
                     /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 ccaccatgcc taagaaaaag aggaaggtgg
     2701 cggcgcctgg atccggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa
     2761 gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg
     2821 acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact
     2881 gtacaggcag tggagacaag aagtacagca tcggcctggc catcggcacc aactctgtgg
     2941 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     3001 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     3061 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     3121 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     3181 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     3241 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     3301 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     3361 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     3421 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     3481 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     3541 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     3601 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     3661 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     3721 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     3781 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     3841 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     3901 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     3961 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     4021 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     4081 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     4141 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     4201 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     4261 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     4321 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     4381 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     4441 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     4501 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     4561 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     4621 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     4681 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     4741 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     4801 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     4861 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     4921 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     4981 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     5041 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     5101 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     5161 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     5221 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     5281 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     5341 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     5401 atgtggacgc catcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     5461 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     5521 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     5581 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     5641 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     5701 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     5761 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     5821 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     5881 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     5941 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     6001 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     6061 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     6121 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     6181 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     6241 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     6301 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     6361 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     6421 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     6481 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     6541 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     6601 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     6661 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     6721 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     6781 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     6841 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     6901 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     6961 agacacggat cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg
     7021 ctggacaggc taagaagaag aaaggatcca ttttcaaacc agaagaacta cgacaggcac
     7081 tgatgccaac tttggaggca ctttaccgtc aggatccaga atcccttccc tttcgtcaac
     7141 ctgtggaccc tcagctttta ggaatccctg attactttga tattgtgaag agccccatgg
     7201 atctttctac cattaagagg aagttagaca ctggacagta tcaggagccc tggcagtatg
     7261 tcgatgatat ttggcttatg ttcaataatg cctggttata taaccggaaa acatcacggg
     7321 tatacaaata ctgctccaag ctctctgagg tctttgaaca agaaattgac ccagtgatgc
     7381 aaagccttgg atactgttgt ggcagaaagt tggagttctc tccacagaca ctgtgttgct
     7441 acggcaaaca gttgtgcaca atacctcgtg atgccactta ttacagttac cagaacaggt
     7501 atcatttctg tgagaagtgt ttcaatgaga tccaagggga gagcgtttct ttgggggatg
     7561 acccttccca gcctcaaact acaataaata aagaacaatt ttccaagaga aaaaatgaca
     7621 cactggatcc tgaactgttt gttgaatgta cagagtgcgg aagaaagatg catcagatct
     7681 gtgtccttca ccatgagatc atctggcctg ctggattcgt ctgtgatggc tgtttaaaga
     7741 aaagtgcacg aactaggaaa gaaaataagt tttctgctaa aaggttgcca tctaccagac
     7801 ttggcacctt tctagagaat cgtgtgaatg actttctgag gcgacagaat caccctgagt
     7861 caggagaggt cactgttaga gtagttcatg cttctgacaa aaccgtggaa gtaaaaccag
     7921 gcatgaaagc aaggtttgtg gacagtggag agatggcaga atcctttcca taccgaacca
     7981 aagccctctt tgcctttgaa gaaattgatg gtgttgacct gtgcttcttt ggcatgcatg
     8041 ttcaagagta tggctctgac tgccctccac ccaaccagag gagagtatac atatcttacc
     8101 tcgatagtgt tcatttcttc cgtcctaaat gcttgaggac tgcagtctat catgaaatcc
     8161 taattggata tttagaatat gtcaagaaat taggttacac aacagggcat atttgggcat
     8221 gtccaccaag tgagggagat gattatatct tccattgcca tcctcctgac cagaagatac
     8281 ccaagcccaa gcgactgcag gaatggtaca aaaaaatgct tgacaaggct gtatcagagc
     8341 gtattgtcca tgactacaag gatattttta aacaagctac tgaagataga ttaacaagtg
     8401 caaaggaatt gccttatttc gagggtgatt tctggcccaa tgttctggaa gaaagcatta
     8461 aggaactgga acaggaggaa gaagagagaa aacgagagga aaacaccagc aatgaaagca
     8521 cagatgtgac caagggagac agcaaaaatg ctaaaaagaa gaataataag aaaaccagca
     8581 aaaataagag cagcctgagt aggggcaaca agaagaaacc cgggatgccc aatgtatcta
     8641 acgacctctc acagaaacta tatgccacca tggagaagca taaagaggtc ttctttgtga
     8701 tccgcctcat tgctggccct gctgccaact ccctgcctcc cattgttgat cctgatcctc
     8761 tcatcccctg cgatctgatg gatggtcggg atgcgtttct cacgctggca agggacaagc
     8821 acctggagtt ctcttcactc cgaagagccc agtggtccac catgtgcatg ctggtggagc
     8881 tgcacacgca gagccaggac ggatccgatt acaaagacga tgacgataag ggatccggcg
     8941 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgg
     9001 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     9061 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     9121 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     9181 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     9241 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     9301 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     9361 aattgctgcc ctctggttat gtgtgggagg gctaaacgcg ttaagtcgac aatcaacctc
     9421 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     9481 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     9541 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     9601 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     9661 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     9721 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     9781 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     9841 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     9901 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     9961 ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac
    10021 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    10081 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
    10141 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
    10201 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
    10261 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
    10321 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
    10381 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
    10441 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
    10501 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
    10561 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
    10621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
    10681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
    10741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
    10801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
    10861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
    10921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
    10981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
    11041 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
    11101 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
    11161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
    11221 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
    11281 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
    11341 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
    11401 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
    11461 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
    11521 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
    11581 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
    11641 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
    11701 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
    11761 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
    11821 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
    11881 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
    11941 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
    12001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
    12061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    12121 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
    12181 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
    12241 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
    12301 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
    12361 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    12421 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    12481 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    12541 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    12601 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    12661 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    12721 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    12781 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    12841 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    12901 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    12961 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    13021 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    13081 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    13141 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    13201 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    13261 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    13321 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    13381 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    13441 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
    13501 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    13561 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    13621 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    13681 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    13741 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    13801 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    13861 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    13921 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    13981 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    14041 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    14101 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    14161 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    14221 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    14281 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
    14341 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
    14401 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
    14461 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
    14521 cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
