LOCUS       sequence-60247-e       10477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a catalytically inactive, human codon-optimized Cas9 
            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 10477)
  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 10477)
  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-humanized
FEATURES             Location/Qualifiers
     source          1..10477
                     /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             5539..5559
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5563..5583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5587..5607
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        5620..6119
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5655..5678)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6052..6071
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6140..6739
                     /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(6140..6159)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6636..6656
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6839..7353
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(7522..7538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7522..7538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7535..7557)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7546..7562
                     /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(7570..7576,7577..7594,7595..7600))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7615..7636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7753..7770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7924..8512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8004..8023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8683..9474,9475..9543))
                     /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     9306..9325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9544..9648)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9716..9734
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9772..9794)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9894..9913
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10107..10129
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10257..10276
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(10476..10477,1..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 gagctgatcc aaaaaagaag agaaaggtag atccaaaaaa gaagagaaag
     5581 gtagatccaa aaaagaagag aaaggtatag aattctaccg ggtaggggag gcgcttttcc
     5641 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     5701 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     5761 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     5821 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     5881 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     5941 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     6001 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     6061 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     6121 cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     6181 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     6241 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     6301 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     6361 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     6421 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     6481 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     6541 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     6601 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     6661 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     6721 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     6781 acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6841 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6901 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     6961 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     7021 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     7081 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     7141 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     7201 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     7261 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     7321 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     7381 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     7441 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aagagctggg
     7501 gagaattaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7561 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7621 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     7681 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8101 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     8161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     8221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     8281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     8341 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     8401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     8461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     8521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     8581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     8641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     8701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     9001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     9061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     9121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     9181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     9241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     9301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     9361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     9421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     9481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     9541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     9601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9661 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     9721 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     9781 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     9841 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9901 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9961 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
    10021 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
    10081 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg
    10141 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
    10201 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
    10261 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
    10321 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
    10381 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
    10441 taaaagattt tatttagtct ccagaaaaag gggggaa
//
