LOCUS       sequence-379929-       14250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding VP64-dCas9-VP64 with 2A Blast 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14250)
  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 14250)
  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-VP64_Blast
FEATURES             Location/Qualifiers
     source          1..14250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2687..3865
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2918..3856
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3813..3833
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3872..3881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3881..4030
                     /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             4046..4093
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(4094..4117,4118..4120,4121..6676,6677..6679,
                     6680..8194)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8249..8269
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8294..8443
                     /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             8465..8518
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             8519..8917
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8942..9530
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8995..9015)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9413..9424)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9533..9549)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9534..9550)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10055..10235
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10261..10278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10267..10491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10537..10965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10624..10643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10834..10855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10974..10994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10979..11308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11159..11294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11221..11240
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11356..11403
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11422..11796
                     /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    11926..12047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11963..11982)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12017..12036
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12096..12112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12096..12112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12109..12131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12120..12136
                     /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(12144..12150,12151..12168,12169..12174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12189..12210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12327..12344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12498..13086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12578..12597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13257..14048,14049..14117))
                     /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     13880..13899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14118..14222)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta gctagctgca aagatggata aagttttaaa cagagaggaa
     2641 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     2701 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2761 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2821 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2881 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     2941 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
     3001 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     3061 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     3121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     3181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     3241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     3301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     3361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     3421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     3481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     3541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     3601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     3661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     3721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     3781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     3841 ttttcttcca tttcaggtgt cgtgacgtac ggccaccatg gacgcattgg acgattttga
     3901 tctggatatg ctgggaagtg acgccctcga tgattttgac cttgacatgc ttggttcgga
     3961 tgcccttgat gactttgacc tcgacatgct cggcagtgac gcccttgatg atttcgacct
     4021 ggacatgctg ggatccgctg gctccaaaag gccggcggcc acgaaaaagg ccggccaggc
     4081 aaaaaagaaa aaggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
     4141 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     4201 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     4261 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     4321 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     4381 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     4441 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     4501 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     4561 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     4621 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     4681 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     4741 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     4801 gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     4861 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     4921 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     4981 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     5041 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     5101 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     5161 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     5221 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     5281 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     5341 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     5401 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     5461 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     5521 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     5581 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     5641 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     5701 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     5761 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     5821 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     5881 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     5941 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     6001 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     6061 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     6121 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     6181 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     6241 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     6301 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     6361 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     6421 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     6481 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     6541 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     6601 tgtggaccac atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     6661 gaccagaagc gacaaggccc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     6721 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     6781 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     6841 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     6901 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     6961 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     7021 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     7081 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     7141 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     7201 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     7261 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     7321 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     7381 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     7441 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     7501 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     7561 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     7621 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     7681 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     7741 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     7801 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     7861 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     7921 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     7981 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     8041 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     8101 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     8161 gacacggatc gacctgtctc agctgggagg cgacagcgct ggaggaggtg gaagcggagg
     8221 aggaggaagc ggaggaggag gtagcggacc taagaaaaag aggaaggtgg cggccgctgg
     8281 atccggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct
     8341 cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat
     8401 gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact gtacaggcag
     8461 tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaat
     8521 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     8581 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     8641 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     8701 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     8761 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     8821 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     8881 tgaattgctg ccctctggtt atgtgtggga gggctaagaa ttcgatatca agcttatcgg
     8941 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     9001 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     9061 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     9121 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     9181 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     9241 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     9301 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     9361 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     9421 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     9481 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     9541 tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     9601 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     9661 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     9721 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     9781 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     9841 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     9901 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     9961 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
    10021 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
    10081 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
    10141 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
    10201 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
    10261 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
    10321 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
    10381 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
    10441 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
    10501 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
    10561 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
    10621 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
    10681 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
    10741 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
    10801 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
    10861 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
    10921 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
    10981 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
    11041 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
    11101 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
    11161 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
    11221 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
    11281 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
    11341 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
    11401 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
    11461 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
    11521 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
    11581 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
    11641 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
    11701 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
    11761 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
    11821 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
    11881 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
    11941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    12001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
    12061 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    12121 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    12181 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    12241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    12301 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    12361 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    12421 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    12481 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    12541 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    12601 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    12661 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    12721 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    12781 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    12841 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    12901 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    12961 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    13021 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    13081 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    13141 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    13201 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    13261 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    13321 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    13381 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    13441 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    13501 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    13561 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    13621 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    13681 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    13741 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    13801 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    13861 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    13921 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    13981 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    14041 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    14101 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    14161 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    14221 cgcacatttc cccgaaaagt gccacctgac
//
