LOCUS       sequence-66611-e       11987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector containing MSCV LTR promoter, dCas9 that is 
            fused to 2x NLS, p65 activation domain and tagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11987)
  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 11987)
  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-p65AD-BFP
FEATURES             Location/Qualifiers
     source          1..11987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1417..1426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1423..1449,1450..1452,1453..3939,3940..3942,
                     3943..5526)
                     /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             5536..5556
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5572..5592
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6022..6393
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             6400..7095
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     promoter        7130..7629
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7165..7188)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7562..7581
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7650..8249
                     /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(7650..7669)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8146..8166
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             8349..8863
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(9032..9048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9032..9048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9045..9067)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9056..9072
                     /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(9080..9086,9087..9104,9105..9110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9125..9146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9263..9280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9434..10022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9514..9533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10193..10984,10985..11053))
                     /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     10816..10835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11054..11158)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11226..11244
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(11282..11304)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     11404..11423
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11617..11639
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11767..11786
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(11986..11987,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctcgcca ccatggacaa gaagtattct
     1441 atcggactgg ccatcgggac taatagcgtc gggtgggccg tgatcactga cgagtacaag
     1501 gtgccctcta agaagttcaa ggtgctcggg aacaccgacc ggcattccat caagaaaaat
     1561 ctgatcggag ctctcctctt tgattcaggg gagaccgctg aagcaacccg cctcaagcgg
     1621 actgctagac ggcggtacac caggaggaag aaccggattt gttaccttca agagatattc
     1681 tccaacgaaa tggcaaaggt cgacgacagc ttcttccata ggctggaaga atcattcctc
     1741 gtggaagagg ataagaagca tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg
     1801 gcctatcacg agaaataccc aaccatctac catcttcgca aaaagctggt ggactcaacc
     1861 gacaaggcag acctccggct tatctacctg gccctggccc acatgatcaa gttcagaggc
     1921 cacttcctga tcgagggcga cctcaatcct gacaatagcg atgtggataa actgttcatc
     1981 cagctggtgc agacttacaa ccagctcttt gaagagaacc ccatcaatgc aagcggagtc
     2041 gatgccaagg ccattctgtc agcccggctg tcaaagagcc gcagacttga gaatcttatc
     2101 gctcagctgc cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc tctttcactt
     2161 gggctgactc ccaatttcaa gtctaatttc gacctggcag aggatgccaa gctgcaactg
     2221 tccaaggaca cctatgatga cgatctcgac aacctcctgg cccagatcgg tgaccaatac
     2281 gccgaccttt tccttgctgc taagaatctt tctgacgcca tcctgctgtc tgacattctc
     2341 cgcgtgaaca ctgaaatcac caaggcccct ctttcagctt caatgattaa gcggtatgat
     2401 gagcaccacc aggacctgac cctgcttaag gcactcgtcc ggcagcagct tccggagaag
     2461 tacaaggaaa tcttctttga ccagtcaaag aatggatacg ccggctacat cgacggaggt
     2521 gcctcccaag aggaatttta taagtttatc aaacctatcc ttgagaagat ggacggcacc
     2581 gaagagctcc tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg cactttcgac
     2641 aatgggagca ttccccacca gatccatctt ggggagcttc acgccatcct tcggcgccaa
     2701 gaggacttct acccctttct taaggacaac agggagaaga ttgagaaaat tctcactttc
     2761 cgcatcccct actacgtggg acccctcgcc agaggaaata gccggtttgc ttggatgacc
     2821 agaaagtcag aagaaactat cactccctgg aacttcgaag aggtggtgga caagggagcc
     2881 agcgctcagt cattcatcga acggatgact aacttcgata agaacctccc caatgagaag
     2941 gtcctgccga aacattccct gctctacgag tactttaccg tgtacaacga gctgaccaag
     3001 gtgaaatatg tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga acaaaagaag
     3061 gcaattgtgg accttctgtt caagaccaat agaaaggtga ccgtgaagca gctgaaggag
     3121 gactatttca agaaaattga atgcttcgac tctgtggaga ttagcggggt cgaagatcgg
     3181 ttcaacgcaa gcctgggtac ctaccatgat ctgcttaaga tcatcaagga caaggatttt
     3241 ctggacaatg aggagaacga ggacatcctt gaggacattg tcctgactct cactctgttc
     3301 gaggaccggg aaatgatcga ggagaggctt aagacctacg cccatctgtt cgacgataaa
     3361 gtgatgaagc aacttaaacg gagaagatat accggatggg gacgccttag ccgcaaactc
     3421 atcaacggaa tccgggacaa acagagcgga aagaccattc ttgatttcct taagagcgac
     3481 ggattcgcta atcgcaactt catgcaactt atccatgatg attccctgac ctttaaggag
     3541 gacatccaga aggcccaagt gtctggacaa ggtgactcac tgcacgagca tatcgcaaat
     3601 ctggctggtt cacccgctat taagaagggt attctccaga ccgtgaaagt cgtggacgag
     3661 ctggtcaagg tgatgggtcg ccataaacca gagaacattg tcatcgagat ggccagggaa
     3721 aaccagacta cccagaaggg acagaagaac agcagggagc ggatgaaaag aattgaggaa
     3781 gggattaagg agctcgggtc acagatcctt aaagagcacc cggtggaaaa cacccagctt
     3841 cagaatgaga agctctatct gtactacctt caaaatggac gcgatatgta tgtggaccaa
     3901 gagcttgata tcaacaggct ctcagactac gacgtggacg ccatcgtccc tcagagcttc
     3961 ctcaaagacg actcaattga caataaggtg ctgactcgct cagacaagaa ccggggaaag
     4021 tcagataacg tgccctcaga ggaagtcgtg aaaaagatga agaactattg gcgccagctt
     4081 ctgaacgcaa agctgatcac tcagcggaag ttcgacaatc tcactaaggc tgagaggggc
     4141 ggactgagcg aactggacaa agcaggattc attaaacggc aacttgtgga gactcggcag
     4201 attactaaac atgtcgccca aatccttgac tcacgcatga ataccaagta cgacgaaaac
     4261 gacaaactta tccgcgaggt gaaggtgatt accctgaagt ccaagctggt cagcgatttc
     4321 agaaaggact ttcaattcta caaagtgcgg gagatcaata actatcatca tgctcatgac
     4381 gcatatctga atgccgtggt gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc
     4441 gagttcgtgt acggagacta caaggtctac gacgtgcgca agatgattgc caaatctgag
     4501 caggagatcg gaaaggccac cgcaaagtac ttcttctaca gcaacatcat gaatttcttc
     4561 aagaccgaaa tcacccttgc aaacggtgag atccggaaga ggccgctcat cgagactaat
     4621 ggggagactg gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg
     4681 ctttctatgc ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct
     4741 aaggaatcaa tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg
     4801 gaccctaaga agtacggcgg attcgattca ccaactgtgg cttattctgt cctggtcgtg
     4861 gctaaggtgg aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact gctgggtatc
     4921 accattatgg agcgcagctc cttcgagaag aacccaattg actttctcga agccaaaggt
     4981 tacaaggaag tcaagaagga ccttatcatc aagctcccaa agtatagcct gttcgaactg
     5041 gagaatgggc ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg taatgagctg
     5101 gctctcccct ccaagtacgt gaatttcctc taccttgcaa gccattacga gaagctgaag
     5161 gggagccccg aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg
     5221 gacgagatca ttgagcagat ttccgagttt tctaaacgcg tcattctcgc tgatgccaac
     5281 ctcgataaag tccttagcgc atacaataag cacagagaca aaccaattcg ggagcaggct
     5341 gagaatatca tccacctgtt caccctcacc aatcttggtg cccctgccgc attcaagtac
     5401 ttcgacacca ccatcgaccg gaaacgctat acctccacca aagaagtgct ggacgccacc
     5461 ctcatccacc agagcatcac cggactttac gaaactcgga ttgacctctc acagctcgga
     5521 ggggatgagg gagctcccaa gaaaaagcgc aaggtaggta gttccggatc tccgaaaaag
     5581 aaacgcaaag ttggtagtca gtatctgccc gacacagatg atagacaccg aatcgaagag
     5641 aaacgcaagc gaacgtatga aaccttcaaa tcgatcatga agaaatcgcc cttctcgggt
     5701 ccgaccgatc ccaggccccc accgagaagg attgcggtcc cgtcccgctc gtcggccagc
     5761 gtgccgaagc ctgcgccgca gccctacccc ttcacgtcga gcctgagcac aatcaattat
     5821 gacgagttcc cgacgatggt gttcccctcg ggacaaatct cacaagcctc ggcgctcgca
     5881 ccagcgcctc cccaagtcct tccgcaagcg cctgccccag cgcctgcacc ggcaatggtg
     5941 tccgccctcg cacaggcccc tgcgcccgtc cccgtgctcg cgcctggacc gccccaggcg
     6001 gtcgctccac cggctccgaa gccgacgcag gccggagagg gaacactctc cgaagcactt
     6061 cttcaactcc agtttgatga cgaggatctt ggagcactcc ttggaaactc gacagaccct
     6121 gcggtgttta ccgacctcgc gtcagtagat aactccgaat ttcagcagct tttgaaccag
     6181 ggtatcccgg tcgcgccaca tacaacggag cccatgttga tggaataccc cgaagcaatc
     6241 acgagacttg tgacgggagc gcagcggcct cccgatcccg cacccgcacc tttgggggca
     6301 cctggcctcc ctaacggact tttgagcggc gacgaggatt tctcctccat cgccgatatg
     6361 gatttctcag ccttgctgtc acagatttcc agcggatcta gcgagctgat taaggagaac
     6421 atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc
     6481 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
     6541 ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc
     6601 ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc
     6661 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     6721 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc
     6781 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac
     6841 cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc
     6901 catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa gaacctcaag
     6961 atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag
     7021 acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg
     7081 gggcacaagc ttaatggatc ctagactctc gagagtctcg aattctaccg ggtaggggag
     7141 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     7201 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     7261 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     7321 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     7381 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     7441 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     7501 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     7561 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     7621 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg
     7681 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     7741 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     7801 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     7861 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     7921 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     7981 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     8041 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     8101 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     8161 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     8221 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     8281 aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa
     8341 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     8401 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     8461 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     8521 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     8581 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     8641 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     8701 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg
     8761 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     8821 tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga
     8881 agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag
     8941 ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag
     9001 aagagctggg gagaattaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     9061 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     9121 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     9181 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     9241 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     9301 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     9361 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     9421 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     9481 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     9541 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     9601 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     9661 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     9721 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     9781 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     9841 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     9901 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9961 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    10021 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    10081 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    10141 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    10201 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    10261 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    10321 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    10381 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    10441 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    10501 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    10561 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    10621 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    10681 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    10741 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    10801 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    10861 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    10921 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10981 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    11041 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    11101 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    11161 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
    11221 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
    11281 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
    11341 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
    11401 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
    11461 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
    11521 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
    11581 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
    11641 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
    11701 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
    11761 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
    11821 ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt
    11881 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
    11941 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaa
//
