LOCUS       sequence-66610-e       11369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector containing MSCV LTR promoter, dCas9 that is 
            fused to 2x NLS, VP64 and tagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11369)
  AUTHORS   Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, 
            Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, 
            Weissman JS, Qi LS
  TITLE     CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in 
            Eukaryotes.
  JOURNAL   Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 
            10.1016/j.cell.2013.06.044.
  PUBMED    23849981
REFERENCE   2  (bases 1 to 11369)
  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: "Cell. 2013 
            Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044."
COMMENT     Sequence Label: pMSCV-LTR-dCas9-VP64-BFP
FEATURES             Location/Qualifiers
     source          1..11369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1419..1428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1425..1451,1452..1454,1455..3941,3942..3944,
                     3945..5528)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5538..5558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5574..5594
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5601..5750
                     /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             5757..5777
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5784..6479
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     promoter        6514..7013
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6549..6572)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6946..6965
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7034..7633
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7034..7053)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7530..7550
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7733..8247
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(8416..8432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8416..8432)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8429..8451)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8440..8456
                     /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(8464..8470,8471..8488,8489..8494))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8509..8530
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8647..8664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8818..9406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8898..8917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9577..10368,10369..10437))
                     /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     10200..10219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10438..10542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10610..10628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10666..10688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10788..10807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11001..11023
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11151..11170
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctcgc caccatggac aagaagtatt
     1441 ctatcggact ggccatcggg actaatagcg tcgggtgggc cgtgatcact gacgagtaca
     1501 aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc atcaagaaaa
     1561 atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc cgcctcaagc
     1621 ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt caagagatat
     1681 tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa gaatcattcc
     1741 tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc gtcgacgagg
     1801 tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg gtggactcaa
     1861 ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc aagttcagag
     1921 gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat aaactgttca
     1981 tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat gcaagcggag
     2041 tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt gagaatctta
     2101 tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt gctctttcac
     2161 ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc aagctgcaac
     2221 tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc ggtgaccaat
     2281 acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg tctgacattc
     2341 tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt aagcggtatg
     2401 atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag cttccggaga
     2461 agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac atcgacggag
     2521 gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag atggacggca
     2581 ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag cgcactttcg
     2641 acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc cttcggcgcc
     2701 aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa attctcactt
     2761 tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt gcttggatga
     2821 ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg gacaagggag
     2881 ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc cccaatgaga
     2941 aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac gagctgacca
     3001 aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc gaacaaaaga
     3061 aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag cagctgaagg
     3121 aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg gtcgaagatc
     3181 ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag gacaaggatt
     3241 ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact ctcactctgt
     3301 tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg ttcgacgata
     3361 aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt agccgcaaac
     3421 tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc cttaagagcg
     3481 acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg acctttaagg
     3541 aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag catatcgcaa
     3601 atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa gtcgtggacg
     3661 agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag atggccaggg
     3721 aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa agaattgagg
     3781 aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa aacacccagc
     3841 ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg tatgtggacc
     3901 aagagcttga tatcaacagg ctctcagact acgacgtgga cgccatcgtc cctcagagct
     3961 tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag aaccggggaa
     4021 agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat tggcgccagc
     4081 ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag gctgagaggg
     4141 gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg gagactcggc
     4201 agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag tacgacgaaa
     4261 acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg gtcagcgatt
     4321 tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat catgctcatg
     4381 acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca aagctggaaa
     4441 gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt gccaaatctg
     4501 agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc atgaatttct
     4561 tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc atcgagacta
     4621 atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc gtgcgcaaag
     4681 tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc ggaggctttt
     4741 ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg aagaaggatt
     4801 gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct gtcctggtcg
     4861 tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa ctgctgggta
     4921 tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc gaagccaaag
     4981 gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc ctgttcgaac
     5041 tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag ggtaatgagc
     5101 tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac gagaagctga
     5161 aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat aagcattatc
     5221 tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc gctgatgcca
     5281 acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt cgggagcagg
     5341 ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc gcattcaagt
     5401 acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg ctggacgcca
     5461 ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc tcacagctcg
     5521 gaggggatga gggagctccc aagaaaaagc gcaaggtagg tagttccgga tctccgaaaa
     5581 agaaacgcaa agttggtagt gatgctttag acgattttga cttagatatg cttggttcag
     5641 acgcgttaga cgacttcgac ctagacatgt taggctcaga tgcattggac gacttcgatt
     5701 tagatatgtt gggctccgat gccctagatg actttgatct agatatgcta ggtagtccca
     5761 aaaagaagag gaaagtggga tctagcgagc tgattaagga gaacatgcac atgaagctgt
     5821 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
     5881 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     5941 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
     6001 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
     6061 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     6121 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
     6181 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
     6241 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca
     6301 tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
     6361 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
     6421 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatg
     6481 gatcctagac tctcgagagt ctcgaattct accgggtagg ggaggcgctt ttcccaaggc
     6541 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     6601 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     6661 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     6721 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     6781 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     6841 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     6901 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     6961 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
     7021 gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     7081 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     7141 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     7201 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     7261 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     7321 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     7381 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     7441 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     7501 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     7561 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     7621 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     7681 ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     7741 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
     7801 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
     7861 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     7921 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     7981 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     8041 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     8101 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc
     8161 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     8221 gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat
     8281 tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc
     8341 acatactcca atactcctga aatagttcat tatggacagc gcagaagagc tggggagaat
     8401 taattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8461 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8521 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     9001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     9061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     9121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     9181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     9241 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     9301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     9361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     9421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     9481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     9541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     9601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     9661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     9721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    10021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    10081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    10141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    10201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    10261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    10321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    10381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    10441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    10501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    10561 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
    10621 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
    10681 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
    10741 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
    10801 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
    10861 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
    10921 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
    10981 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
    11041 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
    11101 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
    11161 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
    11221 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
    11281 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
    11341 attttattta gtctccagaa aaagggggg
//
