LOCUS       sequence-131020-       10076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Streptococcus pyogenes dCas9 (D10A, H840A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10076)
  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 10076)
  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: pCR1003
FEATURES             Location/Qualifiers
     source          1..10076
                     /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..4188
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4198..5298
                     /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     5272..5295
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             5353..5373
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(5380..5406,5407..5409,5410..7896,7897..7899,
                     7900..9486)
                     /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"
     primer_bind     complement(9612..9630)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9626..9673
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(10039..10044,10045..10061,10062..10067))
                     /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 cattgaaaaa 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 agcggtccag 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 caccatcacc atcaccatgg ttcttctatg
     4201 aaaatcgaag aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc
     4261 gctgaagtcg gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg
     4321 gataaactgg aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc
     4381 ttctgggcac acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc
     4441 ccggacaaag cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac
     4501 ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat
     4561 ctgctgccga acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa
     4621 gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg
     4681 attgctgctg acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac
     4741 gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa
     4801 aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc
     4861 gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg
     4921 aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc
     4981 gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc
     5041 gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt
     5101 gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccact
     5161 atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg
     5221 tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc
     5281 ctgaaagacg cgcagactaa ttcgagctcg aacaacaaca acaataacaa taacaacaac
     5341 ctcgggatcg aggaaaacct gtacttccaa tccaatgcaa tggataagaa atactcaata
     5401 ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
     5461 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
     5521 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca
     5581 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
     5641 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
     5701 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
     5761 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
     5821 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
     5881 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
     5941 ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat
     6001 gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct
     6061 cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt
     6121 ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca
     6181 aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct
     6241 gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga
     6301 gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa
     6361 catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat
     6421 aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct
     6481 agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag
     6541 gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac
     6601 ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa
     6661 gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga
     6721 attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg
     6781 aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca
     6841 gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta
     6901 ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc
     6961 aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc
     7021 attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat
     7081 tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt
     7141 aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg
     7201 gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa
     7261 gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg
     7321 atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt
     7381 aatggtatta gggataagca atctggcaaa acaatattag attttttgaa atcagatggt
     7441 tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac
     7501 attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta
     7561 gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg
     7621 gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat
     7681 cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt
     7741 atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa
     7801 aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa
     7861 ttagatatta atcgtttaag tgattatgat gtcgatgcca ttgttccaca aagtttcctt
     7921 aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg
     7981 gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta
     8041 aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt
     8101 ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc
     8161 actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat
     8221 aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga
     8281 aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg
     8341 tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag
     8401 tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa
     8461 gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa
     8521 acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg
     8581 gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg
     8641 tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag
     8701 gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat
     8761 ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct
     8821 aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca
     8881 attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat
     8941 aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa
     9001 aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct
     9061 ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt
     9121 agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat
     9181 gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta
     9241 gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa
     9301 aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt
     9361 gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt
     9421 atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt
     9481 gactaataac attggaagtg gataacggat ccgcgatcgc ggcgcgccac ctggtggccg
     9541 gccggtacca cgcgtgcgcg ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     9601 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     9661 gaggggtttt ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg
     9721 ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc
     9781 caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata
     9841 gcgctagcag cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga
     9901 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga
     9961 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta
    10021 actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaa
//
