LOCUS       sequence-131019-       10124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Streptococcus pyogenes nCas9 (D10A) nickase carrying two 
            C-terminal SV40 NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10124)
  AUTHORS   Richardson CD, Ray GJ, DeWitt MA, Curie GL, Corn JE
  TITLE     Enhancing homology-directed genome editing by catalytically active 
            and inactive CRISPR-Cas9 using asymmetric donor DNA.
  JOURNAL   Nat Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481.
  PUBMED    26789497
REFERENCE   2  (bases 1 to 10124)
  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: "Nat 
            Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481."
COMMENT     Sequence Label: pCR1054
FEATURES             Location/Qualifiers
     source          1..10124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..36)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(209..277,278..1069)
                     /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(427..446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1729..1748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1982..1999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2014..2156
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2082..2101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2242..2264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3907..3926
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4010..4029
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4010..4028
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4117..4139
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4159..4176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4186..5286
                     /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     5260..5283
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             5341..5361
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      5365..5374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5371..5397,5398..5400,5401..9474)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9481..9501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9508..9528
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9660..9678)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9674..9721
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(10087..10092,10093..10109,10110..10115))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc
     3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa
     4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat
     4141 atacatatga aatcttctca ccatcaccat caccatggtt cttctatgaa aatcgaagaa
     4201 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
     4261 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
     4321 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
     4381 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
     4441 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
     4501 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
     4561 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
     4621 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
     4681 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     4741 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     4801 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     4861 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     4921 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     4981 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     5041 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     5101 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     5161 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     5221 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     5281 cagactaatt cgagctcgaa caacaacaac aataacaata acaacaacct cgggatcgag
     5341 gaaaacctgt acttccaatc caatgccacc atggataaga aatactcaat aggcttagct
     5401 atcggcacaa atagcgtcgg atgggcggtg atcactgatg aatataaggt tccgtctaaa
     5461 aagttcaagg ttctgggaaa tacagaccgc cacagtatca aaaaaaatct tataggggct
     5521 cttttatttg acagtggaga gacagcggaa gcgactcgtc tcaaacggac agctcgtaga
     5581 aggtatacac gtcggaagaa tcgtatttgt tatctacagg agattttttc aaatgagatg
     5641 gcgaaagtag atgatagttt ctttcatcga cttgaagagt cttttttggt ggaagaagac
     5701 aagaagcatg aacgtcatcc tatttttgga aatatagtag atgaagttgc ttatcatgag
     5761 aaatatccaa ctatctatca tctgcgaaaa aaattggtag attctactga taaagcggat
     5821 ttgcgcttaa tctatttggc cttagcgcat atgattaagt ttcgtggtca ttttttgatt
     5881 gagggagatt taaatcctga taatagtgat gtggacaaac tatttatcca gttggtacaa
     5941 acctacaatc aattatttga agaaaaccct attaacgcaa gtggagtaga tgctaaagcg
     6001 attctttctg cacgattgag taaatcaaga cgattagaaa atctcattgc tcagctcccc
     6061 ggtgagaaga aaaatggctt atttgggaat ctcattgctt tgtcattggg tttgacccct
     6121 aattttaaat caaattttga tttggcagaa gatgctaaat tacagctttc aaaagatact
     6181 tacgatgatg atttagataa tttattggcg caaattggag atcaatatgc tgatttgttt
     6241 ttggcagcta agaatttatc agatgctatt ttactttcag atatcctaag agtaaatact
     6301 gaaataacta aggctcccct atcagcttca atgattaaac gctacgatga acatcatcaa
     6361 gacttgactc ttttaaaagc tttagttcga caacaacttc cagaaaagta taaagaaatc
     6421 ttttttgatc aatcaaaaaa cggatatgca ggttatattg atgggggagc tagccaagaa
     6481 gaattttata aatttatcaa accaatttta gaaaaaatgg atggtactga ggaattattg
     6541 gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa cggctctatt
     6601 ccccatcaaa ttcacttggg tgagctgcat gctattttga gaagacaaga agacttttat
     6661 ccatttttaa aagacaatcg tgagaagatt gaaaaaatct tgacttttcg aattccttat
     6721 tatgttggtc cattggcgcg tggcaatagt cgttttgcat ggatgactcg gaagtctgaa
     6781 gaaacaatta ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc agctcaatca
     6841 tttattgaac gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt actaccaaaa
     6901 catagtttgc tttatgagta ttttacggtt tataacgaat tgacaaaggt caaatatgtt
     6961 actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc cattgttgat
     7021 ttactcttca aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa
     7081 aaaatagaat gttttgatag tgttgaaatt tcaggagttg aagatagatt taatgcttca
     7141 ttaggtacct accatgattt gctaaaaatt attaaagata aagatttttt ggataatgaa
     7201 gaaaatgaag atatcttaga ggatattgtt ttaacattga ccttatttga agatagggag
     7261 atgattgagg aaagacttaa aacatatgct cacctctttg atgataaggt gatgaaacag
     7321 cttaaacgtc gccgttatac tggttgggga cgtttgtctc gaaaattgat taatggtatt
     7381 agggataagc aatctggcaa aacaatatta gattttttga aatcagatgg ttttgccaat
     7441 cgcaatttta tgcagctgat ccatgatgat agtttgacat ttaaagaaga cattcaaaaa
     7501 gcacaagtgt ctggacaagg cgatagttta catgaacata ttgcaaattt agctggtagc
     7561 cctgctatta aaaaaggtat tttacagact gtaaaagttg ttgatgaatt ggtcaaagta
     7621 atggggcggc ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact
     7681 caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa
     7741 ttaggaagtc agattcttaa agagcatcct gttgaaaata ctcaattgca aaatgaaaag
     7801 ctctatctct attatctcca aaatggaaga gacatgtatg tggaccaaga attagatatt
     7861 aatcgtttaa gtgattatga tgtcgatcac attgttccac aaagtttcct taaagacgat
     7921 tcaatagaca ataaggtctt aacgcgttct gataaaaatc gtggtaaatc ggataacgtt
     7981 ccaagtgaag aagtagtcaa aaagatgaaa aactattgga gacaacttct aaacgccaag
     8041 ttaatcactc aacgtaagtt tgataattta acgaaagctg aacgtggagg tttgagtgaa
     8101 cttgataaag ctggttttat caaacgccaa ttggttgaaa ctcgccaaat cactaagcat
     8161 gtggcacaaa ttttggatag tcgcatgaat actaaatacg atgaaaatga taaacttatt
     8221 cgagaggtta aagtgattac cttaaaatct aaattagttt ctgacttccg aaaagatttc
     8281 caattctata aagtacgtga gattaacaat taccatcatg cccatgatgc gtatctaaat
     8341 gccgtcgttg gaactgcttt gattaagaaa tatccaaaac ttgaatcgga gtttgtctat
     8401 ggtgattata aagtttatga tgttcgtaaa atgattgcta agtctgagca agaaataggc
     8461 aaagcaaccg caaaatattt cttttactct aatatcatga acttcttcaa aacagaaatt
     8521 acacttgcaa atggagagat tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga
     8581 gaaattgtct gggataaagg gcgagatttt gccacagtgc gcaaagtatt gtccatgccc
     8641 caagtcaata ttgtcaagaa aacagaagta cagacaggcg gattctccaa ggagtcaatt
     8701 ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa aagactggga tccaaaaaaa
     8761 tatggtggtt ttgatagtcc aacggtagct tattcagtcc tagtggttgc taaggtggaa
     8821 aaagggaaat cgaagaagtt aaaatccgtt aaagagttac tagggatcac aattatggaa
     8881 agaagttcct ttgaaaaaaa tccgattgac tttttagaag ctaaaggata taaggaagtt
     8941 aaaaaagact taatcattaa actacctaaa tatagtcttt ttgagttaga aaacggtcgt
     9001 aaacggatgc tggctagtgc cggagaatta caaaaaggaa atgagctggc tctgccaagc
     9061 aaatatgtga attttttata tttagctagt cattatgaaa agttgaaggg tagtccagaa
     9121 gataacgaac aaaaacaatt gtttgtggag cagcataagc attatttaga tgagattatt
     9181 gagcaaatca gtgaattttc taagcgtgtt attttagcag atgccaattt agataaagtt
     9241 cttagtgcat ataacaaaca tagagacaaa ccaatacgtg aacaagcaga aaatattatt
     9301 catttattta cgttgacgaa tcttggagct cccgctgctt ttaaatattt tgatacaaca
     9361 attgatcgta aacgatatac gtctacaaaa gaagttttag atgccactct tatccatcaa
     9421 tccatcactg gtctttatga aacacgcatt gatttgagtc agctaggagg tgacgggtca
     9481 cctaagaaaa aacgaaaagt tgaggatcct aaaaagaaac gaaaagttga ttaataacat
     9541 tggaagtgga taacggatcc gcgatcgcgg cgcgccacct ggtggccggc cggtaccacg
     9601 cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     9661 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     9721 gctgaaagga ggaactatat ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt
     9781 agtcgatagt ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg
     9841 acagtgctcc gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca
     9901 cgccatagtg actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta
     9961 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
    10021 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
    10081 taccgcatta aagcttatcg atgataagct gtcaaacatg agaa
//
