LOCUS       sequence-176888-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S867C/N1054C in cysteine-free (C80S/C574S) S. pyogenes Cas9 
            expression plasmid, HNH-2 FRET construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10073)
  AUTHORS   Sternberg SH, LaFrance B, Kaplan M, Doudna JA
  TITLE     Conformational control of DNA target cleavage by CRISPR-Cas9.
  JOURNAL   Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 
            2015 Oct 28.
  PUBMED    26524520
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"; doi: 
            "10.1038/nature15544"; journalName: "Nature"; date: "2015-11-5- 5"; 
            volume: "527"; issue: "7576"; pages: "110-3"
COMMENT     Sequence Label: pSHS248
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(243..263)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(318..1418)
                     /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     complement(321..344)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(1428..1457)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1477..1499
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1587..1606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1588..1606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1690..1709)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3167..3358
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3352..3374)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3460..3599
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3515..3534
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3614..3631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3785..4373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3865..3884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4544..5335,5336..5404))
                     /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     5167..5186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5405..5509)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5577..5595
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5622..5627,5628..5644,5645..5650)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(6016..6063)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6059..6077
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(6203..10073,1..236))
                     /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
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRISYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKCDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLACGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 atacgattct tccgacgtgt ataccttcta cgagctgtcc gtttgagacg agtcgcttcc
       61 gctgtctctc cactgtcaaa taaaagagcc cctataagat tttttttgat actgtggcgg
      121 tctgtatttc ccagaacctt gaacttttta gacggaacct tatattcatc agtgatcacc
      181 gcccatccga cgctatttgt gccgatatct aagcctattg agtatttctt atccattgca
      241 ttggattgga agtacaggtt ttcctcgatc ccgaggttgt tgttattgtt attgttgttg
      301 ttgttcgagc tcgaattagt ctgcgcgtct ttcagggctt catcgacagt ctgacgaccg
      361 ctggcggcgt tgatcaccgc agtacgcacg gcataccaga aagcggacat ctgcgggatg
      421 ttcggcatga tttcaccttt ctgggcgttt tccatagtgg cggcaatacg tggatctttc
      481 gccaactctt cctcgtaaga cttcagcgct acggcaccca gcggtttgtc tttattaacc
      541 gcttccagac cttcatcagt cagcagatag ttttcgagga actcttttgc cagctctttg
      601 ttcggactgg cggcgttaat acctgcgctc agcacgccaa cgaacggttt ggatggttga
      661 cccttgaagg tcggcagtac cgttacacca taattcactt tgctggtgtc gatgttggac
      721 catgcccacg ggccgttgat ggtcatcgct gtttcgcctt tattaaaggc agcttctgcg
      781 atggagtaat cggtgtctgc attcatgtgt ttgtttttaa tcaggtcaac caggaaggtc
      841 agacccgctt ttgcgccagc gttatccacg cccacgtctt taatgtcgta cttgccgttt
      901 tcatacttga acgcataacc cccgtcagca gcaatcagcg gccaggtgaa gtacggttct
      961 tgcaggttga acatcagcgc gctcttacct ttcgctttca gttctttatc cagcgccggg
     1021 atctcttccc aggtttttgg cgggttcggc agcagatctt tgttataaat cagcgataac
     1081 gcttcaacag cgatcgggta agcaatcagc ttgccgttgt aacgtacggc atcccaggta
     1141 aacggataca gcttgtcctg gaacgctttg tccggggtga tttcagccaa caggccagat
     1201 tgagcgtagc caccaaagcg gtcgtgtgcc cagaagataa tgtcagggcc atcgccagtt
     1261 gccgcaacct gtgggaattt ctcttccagt ttatccggat gctcaacggt gactttaatt
     1321 ccggtatctt tctcgaattt cttaccgact tcagcgagac cgttatagcc tttatcgccg
     1381 ttaatccaga ttaccagttt accttcttcg attttcatag aagaaccatg gtgatggtga
     1441 tggtgatggt gatggtgaga agatttcata tgtatatctc cttcttaaag ttaaacaaaa
     1501 ttatttctag aactcgagtc ctgcagggcg gccgcttaat taaagagctc tccggagccg
     1561 gcactagagg gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg
     1621 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1681 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1741 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     1801 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     1861 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaccg atgcccttga
     1921 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     1981 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     2041 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     2101 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     2161 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     2221 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     2281 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     2341 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg
     2401 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
     2461 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
     2521 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
     2581 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     2641 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
     2701 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     2761 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     2821 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     2881 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
     2941 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
     3001 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
     3061 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
     3121 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
     3181 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     3241 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     3301 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3361 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3421 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     3481 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     3541 gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3601 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3661 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3721 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3781 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3901 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3961 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4021 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4081 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4141 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4201 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4261 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4321 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4381 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4441 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4501 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4561 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4621 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4681 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4741 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4801 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     4861 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4921 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4981 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5041 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5101 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5161 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5221 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5281 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5341 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5401 tcatactctt cctttttcaa tgttattgaa gcatttatca gggttattgt ctcatgagcg
     5461 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5521 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5581 ggcgtatcac gaggcccttt cgtcttcaag aattctcatg tttgacagct tatcatcgat
     5641 aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg
     5701 aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata
     5761 ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc
     5821 gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc
     5881 gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt
     5941 ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat atccggatat
     6001 agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     6061 tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc
     6121 cggatcagag cgcacgcgtg gtaccggccg gccaccaggt ggcgcgccgc gatcgcggat
     6181 ccgttatcca cttccaatgt tattagtcac ctcctagctg actcaaatca atgcgtgttt
     6241 cataaagacc agtgatggat tgatggataa gagtggcatc taaaacttct tttgtagacg
     6301 tatatcgttt acgatcaatt gttgtatcaa aatatttaaa agcagcggga gctccaagat
     6361 tcgtcaacgt aaataaatga ataatatttt ctgcttgttc acgtattggt ttgtctctat
     6421 gtttgttata tgcactaaga actttatcta aattggcatc tgctaaaata acacgcttag
     6481 aaaattcact gatttgctca ataatctcat ctaaataatg cttatgctgc tccacaaaca
     6541 attgtttttg ttcgttatct tctggactac ccttcaactt ttcataatga ctagctaaat
     6601 ataaaaaatt cacatatttg cttggcagag ccagctcatt tcctttttgt aattctccgg
     6661 cactagccag catccgttta cgaccgtttt ctaactcaaa aagactatat ttaggtagtt
     6721 taatgattaa gtctttttta acttccttat atcctttagc ttctaaaaag tcaatcggat
     6781 ttttttcaaa ggaacttctt tccataattg tgatccctag taactcttta acggatttta
     6841 acttcttcga tttccctttt tccaccttag caaccactag gactgaataa gctaccgttg
     6901 gactatcaaa accaccatat ttttttggat cccagtcttt tttacgagca ataagcttgt
     6961 ccgaatttct ttttggtaaa attgactcct tggagaatcc gcctgtctgt acttctgttt
     7021 tcttgacaat attgacttgg ggcatggaca atactttgcg cactgtggca aaatctcgcc
     7081 ctttatccca gacaatttct ccagtttccc cattagtttc gattagaggg cgtttgcgaa
     7141 tctctccaca tgcaagtgta atttctgttt tgaagaagtt catgatatta gagtaaaaga
     7201 aatattttgc ggttgctttg cctatttctt gctcagactt agcaatcatt ttacgaacat
     7261 cataaacttt ataatcacca tagacaaact ccgattcaag ttttggatat ttcttaatca
     7321 aagcagttcc aacgacggca tttagatacg catcatgggc atgatggtaa ttgttaatct
     7381 cacgtacttt atagaattgg aaatcttttc ggaagtcaga aactaattta gattttaagg
     7441 taatcacttt aacctctcga ataagtttat cattttcatc gtatttagta ttcatgcgac
     7501 tatccaaaat ttgtgccaca tgcttagtga tttggcgagt ttcaaccaat tggcgtttga
     7561 taaaaccagc tttatcaagt tcactcaaac ctccacgttc agctttcgtt aaattatcaa
     7621 acttacgttg agtgattaac ttggcgttta gaagttgtct ccaatagttt ttcatctttt
     7681 tgactacttc ttcacttgga acgttatcgc atttaccacg atttttatca gaacgcgtta
     7741 agaccttatt gtctattgaa tcgtctttaa ggaaactttg tggaacaatg tgatcgacat
     7801 cataatcact taaacgatta atatctaatt cttggtccac atacatgtct cttccatttt
     7861 ggagataata gagatagagc ttttcatttt gcaattgagt attttcaaca ggatgctctt
     7921 taagaatctg acttcctaat tctttgatac cttcttcgat tcgtttcata cgctctcgcg
     7981 aatttttctg gcccttttga gttgtctgat tttcacgtgc catttcaata acgatatttt
     8041 ctggcttatg ccgccccatt actttgacca attcatcaac aacttttaca gtctgtaaaa
     8101 tacctttttt aatagcaggg ctaccagcta aatttgcaat atgttcatgt aaactatcgc
     8161 cttgtccaga cacttgtgct ttttgaatgt cttctttaaa tgtcaaacta tcatcatgga
     8221 tcagctgcat aaaattgcga ttggcaaaac catctgattt caaaaaatct aatattgttt
     8281 tgccagattg cttatcccta ataccattaa tcaattttcg agacaaacgt ccccaaccag
     8341 tataacggcg acgtttaagc tgtttcatca ccttatcatc aaagaggtga gcatatgttt
     8401 taagtctttc ctcaatcatc tccctatctt caaataaggt caatgttaaa acaatatcct
     8461 ctaagatatc ttcattttct tcattatcca aaaaatcttt atctttaata atttttagca
     8521 aatcatggta ggtacctaat gaagcattaa atctatcttc aactcctgaa atttcaacac
     8581 tatcaaaaga ttctattttt ttgaaataat cttcttttaa ttgcttaacg gttacttttc
     8641 gatttgtttt gaagagtaaa tcaacaatgg ctttcttctg ttcacctgaa agaaatgctg
     8701 gttttcgcat tccttcagta acatatttga cctttgtcaa ttcgttataa accgtaaaat
     8761 actcataaag caaactatgt tttggtagta ctttttcatt tggaagattt ttatcaaagt
     8821 ttgtcatgcg ttcaataaat gattgagctg aagcaccttt atcgacaact tcttcaaaat
     8881 tccatggggt aattgtttct tcagacttcc gagtcatcca tgcaaaacga ctattgccac
     8941 gcgccaatgg accaacataa taaggaattc gaaaagtcaa gattttttca atcttctcac
     9001 gattgtcttt taaaaatgga taaaagtctt cttgtcttct caaaatagca tgcagctcac
     9061 ccaagtgaat ttgatgggga atagagccgt tgtcaaaggt ccgttgcttg cgcagcaaat
     9121 cttcacgatt tagtttcacc aataattcct cagtaccatc cattttttct aaaattggtt
     9181 tgataaattt ataaaattct tcttggctag ctcccccatc aatataacct gcatatccgt
     9241 tttttgattg atcaaaaaag atttctttat acttttctgg aagttgttgt cgaactaaag
     9301 cttttaaaag agtcaagtct tgatgatgtt catcgtagcg tttaatcatt gaagctgata
     9361 ggggagcctt agttatttca gtatttactc ttaggatatc tgaaagtaaa atagcatctg
     9421 ataaattctt agctgccaaa aacaaatcag catattgatc tccaatttgc gccaataaat
     9481 tatctaaatc atcatcgtaa gtatcttttg aaagctgtaa tttagcatct tctgccaaat
     9541 caaaatttga tttaaaatta ggggtcaaac ccaatgacaa agcaatgaga ttcccaaata
     9601 agccattttt cttctcaccg gggagctgag caatgagatt ttctaatcgt cttgatttac
     9661 tcaatcgtgc agaaagaatc gctttagcat ctactccact tgcgttaata gggttttctt
     9721 caaataattg attgtaggtt tgtaccaact ggataaatag tttgtccaca tcactattat
     9781 caggatttaa atctccctca atcaaaaaat gaccacgaaa cttaatcata tgcgctaagg
     9841 ccaaatagat taagcgcaaa tccgctttat cagtagaatc taccaatttt tttcgcagat
     9901 gatagatagt tggatatttc tcatgataag caacttcatc tactatattt ccaaaaatag
     9961 gatgacgttc atgcttcttg tcttcttcca ccaaaaaaga ctcttcaagt cgatgaaaga
    10021 aactatcatc tactttcgcc atctcatttg aaaaaatctc ctgtagataa gaa
//
