LOCUS       sequence-351748-       10070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of SpCas9 KES(1107-1109)GG (phosphate lock 
            loop mutant) with an N-terminal His-MBP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10070)
  AUTHORS   Cofsky JC, Soczek KM, Knott GJ, Nogales E, Doudna JA
  TITLE     CRISPR-Cas9 bends and twists DNA to read its sequence.
  JOURNAL   Nat Struct Mol Biol. 2022 Apr;29(4):395-402. doi: 
            10.1038/s41594-022-00756-0. Epub 2022 Apr 14.
  PUBMED    35422516
REFERENCE   2  (bases 1 to 10070)
  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/s41594-022-00756-0"; journalName: "Nat Struct Mol Biol"; 
            date: "2022-04"; volume: "29"; issue: "4"; pages: "395-402"
COMMENT     Sequence Label: pJCC_162 SpCas9 KES(1107-1109)GG
FEATURES             Location/Qualifiers
     source          1..10070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     349..368
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     452..471
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        452..470
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             559..581
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             601..630
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             640..1740
                     /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     1714..1737
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1795..1815
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1822..5925
                     /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
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSG
                     GILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6051..6069)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6065..6112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6478..6483,6484..6500,6501..6506))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6533..6551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6619..6723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6724..6792,6793..7584)
                     /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(6942..6961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7755..8343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8244..8263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8497..8514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8529..8668
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8594..8613)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8754..8776
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8770..8961)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
       61 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
      121 gactgggttg aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat
      181 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
      241 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
      301 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
      361 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      421 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
      481 cggtttccct ctagtgccgg ctccggagag ctctttaatt aagcggccgc cctgcaggac
      541 tcgagttcta gaaataattt tgtttaactt taagaaggag atatacatat gaaatcttct
      601 caccatcacc atcaccatca ccatcaccat ggttcttcta tgaaaatcga agaaggtaaa
      661 ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa
      721 ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa
      781 ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc
      841 tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag
      901 gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac
      961 ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca
     1021 aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg
     1081 ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt
     1141 tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct
     1201 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca
     1261 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
     1321 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
     1381 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
     1441 aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact
     1501 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
     1561 tacgaggaag agttggcgaa agatccacgt attgccgcca ctatggaaaa cgcccagaaa
     1621 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
     1681 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact
     1741 aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat cgaggaaaac
     1801 ctgtacttcc aatccaatgc aatggataag aaatactcaa taggcttaga tatcggcaca
     1861 aatagcgtcg gatgggcggt gatcactgat gaatataagg ttccgtctaa aaagttcaag
     1921 gttctgggaa atacagaccg ccacagtatc aaaaaaaatc ttataggggc tcttttattt
     1981 gacagtggag agacagcgga agcgactcgt ctcaaacgga cagctcgtag aaggtataca
     2041 cgtcggaaga atcgtatttg ttatctacag gagatttttt caaatgagat ggcgaaagta
     2101 gatgatagtt tctttcatcg acttgaagag tcttttttgg tggaagaaga caagaagcat
     2161 gaacgtcatc ctatttttgg aaatatagta gatgaagttg cttatcatga gaaatatcca
     2221 actatctatc atctgcgaaa aaaattggta gattctactg ataaagcgga tttgcgctta
     2281 atctatttgg ccttagcgca tatgattaag tttcgtggtc attttttgat tgagggagat
     2341 ttaaatcctg ataatagtga tgtggacaaa ctatttatcc agttggtaca aacctacaat
     2401 caattatttg aagaaaaccc tattaacgca agtggagtag atgctaaagc gattctttct
     2461 gcacgattga gtaaatcaag acgattagaa aatctcattg ctcagctccc cggtgagaag
     2521 aaaaatggct tatttgggaa tctcattgct ttgtcattgg gtttgacccc taattttaaa
     2581 tcaaattttg atttggcaga agatgctaaa ttacagcttt caaaagatac ttacgatgat
     2641 gatttagata atttattggc gcaaattgga gatcaatatg ctgatttgtt tttggcagct
     2701 aagaatttat cagatgctat tttactttca gatatcctaa gagtaaatac tgaaataact
     2761 aaggctcccc tatcagcttc aatgattaaa cgctacgatg aacatcatca agacttgact
     2821 cttttaaaag ctttagttcg acaacaactt ccagaaaagt ataaagaaat cttttttgat
     2881 caatcaaaaa acggatatgc aggttatatt gatgggggag ctagccaaga agaattttat
     2941 aaatttatca aaccaatttt agaaaaaatg gatggtactg aggaattatt ggtgaaacta
     3001 aatcgtgaag atttgctgcg caagcaacgg acctttgaca acggctctat tccccatcaa
     3061 attcacttgg gtgagctgca tgctattttg agaagacaag aagactttta tccattttta
     3121 aaagacaatc gtgagaagat tgaaaaaatc ttgacttttc gaattcctta ttatgttggt
     3181 ccattggcgc gtggcaatag tcgttttgca tggatgactc ggaagtctga agaaacaatt
     3241 accccatgga attttgaaga agttgtcgat aaaggtgctt cagctcaatc atttattgaa
     3301 cgcatgacaa actttgataa aaatcttcca aatgaaaaag tactaccaaa acatagtttg
     3361 ctttatgagt attttacggt ttataacgaa ttgacaaagg tcaaatatgt tactgaagga
     3421 atgcgaaaac cagcatttct ttcaggtgaa cagaagaaag ccattgttga tttactcttc
     3481 aaaacaaatc gaaaagtaac cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa
     3541 tgttttgata gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc attaggtacc
     3601 taccatgatt tgctaaaaat tattaaagat aaagattttt tggataatga agaaaatgaa
     3661 gatatcttag aggatattgt tttaacattg accttatttg aagataggga gatgattgag
     3721 gaaagactta aaacatatgc tcacctcttt gatgataagg tgatgaaaca gcttaaacgt
     3781 cgccgttata ctggttgggg acgtttgtct cgaaaattga ttaatggtat tagggataag
     3841 caatctggca aaacaatatt agattttttg aaatcagatg gttttgccaa tcgcaatttt
     3901 atgcagctga tccatgatga tagtttgaca tttaaagaag acattcaaaa agcacaagtg
     3961 tctggacaag gcgatagttt acatgaacat attgcaaatt tagctggtag ccctgctatt
     4021 aaaaaaggta ttttacagac tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg
     4081 cataagccag aaaatatcgt tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc
     4141 cagaaaaatt cgcgagagcg tatgaaacga atcgaagaag gtatcaaaga attaggaagt
     4201 cagattctta aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa gctctatctc
     4261 tattatctcc aaaatggaag agacatgtat gtggaccaag aattagatat taatcgttta
     4321 agtgattatg atgtcgatca cattgttcca caaagtttcc ttaaagacga ttcaatagac
     4381 aataaggtct taacgcgttc tgataaaaat cgtggtaaat cggataacgt tccaagtgaa
     4441 gaagtagtca aaaagatgaa aaactattgg agacaacttc taaacgccaa gttaatcact
     4501 caacgtaagt ttgataattt aacgaaagct gaacgtggag gtttgagtga acttgataaa
     4561 gctggtttta tcaaacgcca attggttgaa actcgccaaa tcactaagca tgtggcacaa
     4621 attttggata gtcgcatgaa tactaaatac gatgaaaatg ataaacttat tcgagaggtt
     4681 aaagtgatta ccttaaaatc taaattagtt tctgacttcc gaaaagattt ccaattctat
     4741 aaagtacgtg agattaacaa ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt
     4801 ggaactgctt tgattaagaa atatccaaaa cttgaatcgg agtttgtcta tggtgattat
     4861 aaagtttatg atgttcgtaa aatgattgct aagtctgagc aagaaatagg caaagcaacc
     4921 gcaaaatatt tcttttactc taatatcatg aacttcttca aaacagaaat tacacttgca
     4981 aatggagaga ttcgcaaacg ccctctaatc gaaactaatg gggaaactgg agaaattgtc
     5041 tgggataaag ggcgagattt tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat
     5101 attgtcaaga aaacagaagt acagacaggc ggattctccg ggggaatttt accaaaaaga
     5161 aattcggaca agcttattgc tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt
     5221 gatagtccaa cggtagctta ttcagtccta gtggttgcta aggtggaaaa agggaaatcg
     5281 aagaagttaa aatccgttaa agagttacta gggatcacaa ttatggaaag aagttccttt
     5341 gaaaaaaatc cgattgactt tttagaagct aaaggatata aggaagttaa aaaagactta
     5401 atcattaaac tacctaaata tagtcttttt gagttagaaa acggtcgtaa acggatgctg
     5461 gctagtgccg gagaattaca aaaaggaaat gagctggctc tgccaagcaa atatgtgaat
     5521 tttttatatt tagctagtca ttatgaaaag ttgaagggta gtccagaaga taacgaacaa
     5581 aaacaattgt ttgtggagca gcataagcat tatttagatg agattattga gcaaatcagt
     5641 gaattttcta agcgtgttat tttagcagat gccaatttag ataaagttct tagtgcatat
     5701 aacaaacata gagacaaacc aatacgtgaa caagcagaaa atattattca tttatttacg
     5761 ttgacgaatc ttggagctcc cgctgctttt aaatattttg atacaacaat tgatcgtaaa
     5821 cgatatacgt ctacaaaaga agttttagat gccactctta tccatcaatc catcactggt
     5881 ctttatgaaa cacgcattga tttgagtcag ctaggaggtg actaataaca ttggaagtgg
     5941 ataacggatc cgcgatcgcg gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc
     6001 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     6061 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     6121 aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag
     6181 tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc
     6241 cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt
     6301 gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa
     6361 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
     6421 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
     6481 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
     6541 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     6601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataacattg aaaaaggaag
     6721 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6781 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6841 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6901 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6961 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7021 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7081 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7141 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7201 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7261 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7321 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7381 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7441 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7501 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7561 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7621 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7681 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7741 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7801 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7861 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     7921 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7981 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8041 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8101 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8161 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8221 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8281 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8341 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8401 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8461 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8521 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     8581 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     8641 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     8701 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     8761 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     8821 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     8881 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     8941 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     9001 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     9061 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     9121 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     9181 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     9241 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     9301 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     9361 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     9421 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     9481 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     9541 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     9601 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     9661 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     9721 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     9781 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     9841 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     9901 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     9961 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
    10021 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
//
