LOCUS       sequence-220462-       12849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Lentiviral vector expressing Lentiviral
            vector expressing dCas9VP64 under the control of the long human EF1a
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12849)
  AUTHORS   Chong ZS, Ohnishi S, Yusa K, Wright GJ
  TITLE     Pooled extracellular receptor-ligand interaction screening using 
            CRISPR activation.
  JOURNAL   Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3.
  PUBMED    30477585
REFERENCE   2  (bases 1 to 12849)
  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: "Genome 
            Biol."; date: "2018-11-26"; volume: "19(1):205. doi"; pages: " 
            10.1186/s13059-018-1581-3"
COMMENT     Sequence Label: pKLV2-EF1adCas9VP64T2ABsd-W
FEATURES             Location/Qualifiers
     source          1..12849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1212..1259
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(1260..1283,1284..1286,1287..3842,3843..3845,
                     3846..5360)
                     /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             5415..5435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5460..5609
                     /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             5631..5684
                     /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             5685..6083
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(6149..6169)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6567..6578)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7030..7047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7036..7260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7286..7306)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7286..7304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7325..7341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7325..7341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7338..7360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7349..7365
                     /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(7373..7379,7380..7397,7398..7403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7418..7439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7556..7573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7727..8315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7807..7826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8486..9277,9278..9346))
                     /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     9109..9128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9347..9451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9477..9932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9614..9635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9826..9845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10059..10081
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10073..10090
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10074..10090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10100..10119
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10100..10118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        10434..10632
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10587..10607
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10650..10830
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10877..11002
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11501..11734
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11919..11963
                     /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    12261..12378
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(12849,1..1181)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaagtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt ggaagcttgg cgcgcccgta
     1201 cggccaccat gaaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     1261 acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca
     1321 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     1381 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     1441 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     1501 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1561 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1621 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1681 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     1741 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     1801 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     1861 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     1921 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga
     1981 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     2041 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     2101 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     2161 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     2221 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     2281 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     2341 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     2401 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     2461 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     2521 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2581 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2641 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     2701 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     2761 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     2821 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     2881 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     2941 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     3001 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     3061 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     3121 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     3181 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     3241 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     3301 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     3361 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     3421 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     3481 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3541 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3601 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3661 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     3721 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccacatcg
     3781 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     3841 aggcccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     3901 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     3961 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     4021 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     4081 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     4141 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     4201 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     4261 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     4321 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     4381 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     4441 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     4501 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4561 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     4621 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4681 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     4741 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     4801 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     4861 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     4921 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     4981 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     5041 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     5101 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     5161 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     5221 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     5281 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     5341 tgtctcagct gggaggcgac agcgctggag gaggtggaag cggaggagga ggaagcggag
     5401 gaggaggtag cggacctaag aaaaagagga aggtggcggc cgctggatcc ggacgggctg
     5461 acgcattgga cgattttgat ctggatatgc tgggaagtga cgccctcgat gattttgacc
     5521 ttgacatgct tggttcggat gcccttgatg actttgacct cgacatgctc ggcagtgacg
     5581 cccttgatga tttcgacctg gacatgctga ttaactgtac aggcagtgga gagggcagag
     5641 gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccaatggcc aagcctttgt
     5701 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     5761 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     5821 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     5881 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     5941 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     6001 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     6061 ctggttatgt gtgggagggc taagaattcg cggccgctca acctctggat tacaaaattt
     6121 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     6181 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     6241 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     6301 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     6361 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     6421 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     6481 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     6541 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6601 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     6661 tctccctttg ggccgcctcc ccgcaggtac ctttaagacc aatgacttac aaggcagctg
     6721 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaaa
     6781 gaagtcaaga tctgcttttt gcctgtactg ggtctctctg gttagaccag agtctctctg
     6841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtttaaaccc
     7021 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     7081 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     7141 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     7201 gcaaggggga ggattgggaa gtcaatagca ggcatgctgg ggatgcggtg ggctctatgg
     7261 gcggccgtta attaagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     7321 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7381 gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7441 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7561 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7621 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8461 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8521 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8581 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8641 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8701 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8761 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8821 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8881 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8941 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9001 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9061 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9121 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9181 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9241 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9301 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9361 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9421 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     9481 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     9541 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     9601 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     9661 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     9721 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     9781 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     9841 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     9901 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     9961 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
    10021 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
    10081 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgacta gtacataact
    10141 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
    10201 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
    10261 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
    10321 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
    10381 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
    10441 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
    10501 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
    10561 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
    10621 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
    10681 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    10741 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    10801 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
    10861 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
    10921 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
    10981 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
    11041 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
    11101 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
    11161 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
    11221 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
    11281 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
    11341 ggccggccgc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
    11401 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
    11461 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
    11521 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
    11581 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
    11641 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
    11701 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
    11761 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
    11821 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
    11881 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
    11941 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
    12001 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
    12061 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
    12121 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
    12181 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
    12241 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
    12301 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
    12361 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttagta
    12421 ccgggcccta cgcgtgaggg gcgcggaagc cggggcggag gaggagccaa ccccgggctg
    12481 ggctgagacc cgcagaggaa gacgctctag ggatttgtcc cggactagcg agatggcaag
    12541 gctgaggacg ggaggctgat tgagaggcga aggtacaccc taatctcaat acaacctttg
    12601 gagctaagcc agcaatggta gagggaagat tctgcacgtc ccttccaggc ggcctccccg
    12661 tcaccacccc ccccaacccg ccccgaccgg agctgagagt aattcataca aaaggactcg
    12721 cccctgcctt ggggaatccc agggaccgtc gttaaactcc cactaacgta gaacccagag
    12781 atcgctgcgt tcccgccccc tcacccgccc gctctcgtca tcactgaggt ggagaagagc
    12841 atgcgtgag
//
