LOCUS       sequence-197212-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SpCas9 Cluster 2 + Q926A variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10073)
  AUTHORS   Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington 
            LB, Sternberg SH, Joung JK, Yildiz A, Doudna JA
  TITLE     Enhanced proofreading governs CRISPR-Cas9 targeting accuracy.
  JOURNAL   Nature. 2017 Sep 20. doi: 10.1038/nature24268.
  PUBMED    28931002
REFERENCE   2  (bases 1 to 10073)
  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: "Nature. 
            2017 Sep 20. doi: 10.1038/nature24268."
COMMENT     Sequence Label: pJSC197 - Bacterial expression plasmid for SpC...
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1414..1433
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1517..1536
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1517..1535
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1624..1646
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1666..1695
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1705..2805
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2779..2802
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2860..2880
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2887..6993
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISAAAARFAASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(7119..7137)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7133..7180
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7546..7551,7552..7568,7569..7574))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(7601..7619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7687..7791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7792..7860,7861..8652)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8010..8029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8823..9411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9312..9331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9565..9582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9597..9736
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9662..9681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9822..9844
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9838..10029)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
       61 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
      121 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      181 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
      241 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
      301 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
      361 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
      421 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
      481 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
      541 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
      601 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
      661 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
      721 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
      781 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
      841 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
      901 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
      961 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     1021 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     1081 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     1141 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     1201 tctcaagggc atcggtcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     1261 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     1321 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     1381 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     1441 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     1501 ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctagt
     1561 gccggctccg gagagctctt taattaagcg gccgccctgc aggactcgag ttctagaaat
     1621 aattttgttt aactttaaga aggagatata catatgaaat cttctcacca tcaccatcac
     1681 catcaccatc accatggttc ttctatgaaa atcgaagaag gtaaactggt aatctggatt
     1741 aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc
     1801 ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg
     1861 gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct
     1921 caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg
     1981 tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa
     2041 gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag
     2101 atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg
     2161 caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat
     2221 gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt
     2281 ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc
     2341 atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca
     2401 tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag
     2461 ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg
     2521 aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa
     2581 gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg
     2641 gcgaaagatc cacgtattgc cgccactatg gaaaacgccc agaaaggtga aatcatgccg
     2701 aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc
     2761 agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaattc gagctcgaac
     2821 aacaacaaca ataacaataa caacaacctc gggatcgagg aaaacctgta cttccaatcc
     2881 aatgcaatgg ataagaaata ctcaataggc ttagatatcg gcacaaatag cgtcggatgg
     2941 gcggtgatca ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca
     3001 gaccgccaca gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca
     3061 gcggaagcga ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt
     3121 atttgttatc tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt
     3181 catcgacttg aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt
     3241 tttggaaata tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg
     3301 cgaaaaaaat tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta
     3361 gcgcatatga ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat
     3421 agtgatgtgg acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa
     3481 aaccctatta acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa
     3541 tcaagacgat tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt
     3601 gggaatctca ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg
     3661 gcagaagatg ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta
     3721 ttggcgcaaa ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat
     3781 gctattttac tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca
     3841 gcttcaatga ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta
     3901 gttcgacaac aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga
     3961 tatgcaggtt atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca
     4021 attttagaaa aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg
     4081 ctgcgcaagc aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag
     4141 ctgcatgcta ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag
     4201 aagattgaaa aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc
     4261 aatagtcgtt ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt
     4321 gaagaagttg tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt
     4381 gataaaaatc ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt
     4441 acggtttata acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca
     4501 tttctttcag gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa
     4561 gtaaccgtta agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt
     4621 gaaatttcag ctgctgctgc tagatttgct gcttcattag gtacctacca tgatttgcta
     4681 aaaattatta aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat
     4741 attgttttaa cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca
     4801 tatgctcacc tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt
     4861 tggggacgtt tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca
     4921 atattagatt ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat
     4981 gatgatagtt tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat
     5041 agtttacatg aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta
     5101 cagactgtaa aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat
     5161 atcgttattg aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga
     5221 gagcgtatga aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag
     5281 catcctgttg aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat
     5341 ggaagagaca tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc
     5401 gatcacattg ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg
     5461 cgttctgata aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag
     5521 atgaaaaact attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat
     5581 aatttaacga aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa
     5641 cgccaattgg ttgaaactcg cgctatcact aagcatgtgg cacaaatttt ggatagtcgc
     5701 atgaatacta aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta
     5761 aaatctaaat tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt
     5821 aacaattacc atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt
     5881 aagaaatatc caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt
     5941 cgtaaaatga ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt
     6001 tactctaata tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc
     6061 aaacgccctc taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga
     6121 gattttgcca cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca
     6181 gaagtacaga caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag
     6241 cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg
     6301 gtagcttatt cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa
     6361 tccgttaaag agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg
     6421 attgactttt tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta
     6481 cctaaatata gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga
     6541 gaattacaaa aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta
     6601 gctagtcatt atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt
     6661 gtggagcagc ataagcatta tttagatgag attattgagc aaatcagtga attttctaag
     6721 cgtgttattt tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga
     6781 gacaaaccaa tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt
     6841 ggagctcccg ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct
     6901 acaaaagaag ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca
     6961 cgcattgatt tgagtcagct aggaggtgac taataacatt ggaagtggat aacggatccg
     7021 cgatcgcggc gcgccacctg gtggccggcc ggtaccacgc gtgcgcgctg atccggctgc
     7081 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     7141 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     7201 cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
     7261 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
     7321 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
     7381 tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     7441 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     7501 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
     7561 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt
     7621 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7741 atgagacaat aaccctgata aatgcttcaa taacattgaa aaaggaagag tatgagtatt
     7801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     8041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     8341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8941 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     9001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     9421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     9481 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     9541 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     9601 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     9661 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     9721 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     9781 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     9841 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     9901 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     9961 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
    10021 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tga
//
