LOCUS       sequence-60248-e       11170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a catalytically inactive, human codon-optimized 
            Cas9-BFP fusion under the control of Murine Stem Cell retroVirus LTR
            promoter for mammalian gene knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11170)
  AUTHORS   Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim 
            WA
  TITLE     Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific 
            Control of Gene Expression.
  JOURNAL   Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.
  PUBMED    23452860
REFERENCE   2  (bases 1 to 11170)
  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"; doi: 
            "10.1016/j.cell.2013.02"; journalName: "Cell"; date: "2013-02-28- 
            28"; volume: "152"; issue: "5"; pages: "1173-83"
COMMENT     Sequence Label: pdCas9::BFP-humanized
FEATURES             Location/Qualifiers
     source          1..11170
                     /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             5566..6261
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             6271..6291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        6313..6812
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6348..6371)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6745..6764
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6833..7432
                     /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(6833..6852)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7329..7349
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7532..8046
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(8215..8231)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8215..8231)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8228..8250)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8239..8255
                     /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(8263..8269,8270..8287,8288..8293))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8308..8329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8446..8463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8617..9205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8697..8716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9376..10167,10168..10236))
                     /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     9999..10018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10237..10341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10409..10427
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10465..10487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10587..10606
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10800..10822
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10950..10969
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(11169..11170,1..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 gttccagcga gctgattaag
     5581 gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc
     5641 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     5701 gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc
     5761 aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag
     5821 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     5881 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     5941 acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg
     6001 ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc
     6061 gggagccatc tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac
     6121 ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac
     6181 aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc
     6241 aaactggggc acaagcttaa tgagggagct ccaaagaaga agcgtaaggt aggtagttcc
     6301 tagaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     6361 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     6421 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     6481 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     6541 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     6601 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     6661 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     6721 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     6781 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga
     6841 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     6901 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     6961 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     7021 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     7081 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     7141 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     7201 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     7261 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     7321 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     7381 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     7441 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     7501 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     7561 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     7621 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     7681 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     7741 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     7801 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     7861 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     7921 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     7981 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     8041 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata
     8101 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa
     8161 atagttcatt atggacagcg cagaagagct ggggagaatt aattcgtaat catggtcata
     8221 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     8281 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     8341 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     8401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     8461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     8521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     9001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     9061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     9121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     9181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     9241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     9361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     9421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     9481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    10021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    10081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    10141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    10201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    10261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
    10381 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
    10441 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
    10501 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
    10561 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
    10621 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
    10681 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
    10741 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
    10801 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
    10861 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
    10921 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
    10981 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
    11041 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
    11101 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
    11161 aaggggggaa
//
