pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.

LOCUS       pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant               10073 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant.
ACCESSION   .
VERSION     .
KEYWORDS    pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10073
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     393..412
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     496..515
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        496..514
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             603..625
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             645..674
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             684..1784
                     /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     1758..1781
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1839..1859
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1866..5972
                     /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
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6098..6116)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6112..6159
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6525..6553)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6580..6598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6666..6770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6771..7631
                     /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(6989..7008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7802..8390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8291..8310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8544..8561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8576..8715
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8641..8660)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8801..8823
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8817..9008)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
       61 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
      121 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
      181 ctcaagggca tcggtcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
      241 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
      301 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
      361 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      421 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
      481 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagtg
      541 ccggctccgg agagctcttt aattaagcgg ccgccctgca ggactcgagt tctagaaata
      601 attttgttta actttaagaa ggagatatac atatgaaatc ttctcaccat caccatcacc
      661 atcaccatca ccatggttct tctatgaaaa tcgaagaagg taaactggta atctggatta
      721 acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg
      781 gaattaaagt caccgttgag catccggata aactggaaga gaaattccca caggttgcgg
      841 caactggcga tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc
      901 aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt
      961 ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag
     1021 cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga
     1081 tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc
     1141 aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg
     1201 aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc
     1261 tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca
     1321 tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat
     1381 ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg
     1441 gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga
     1501 acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag
     1561 cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg
     1621 cgaaagatcc acgtattgcc gccactatgg aaaacgccca gaaaggtgaa atcatgccga
     1681 acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca
     1741 gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg agctcgaaca
     1801 acaacaacaa taacaataac aacaacctcg ggatcgagga aaacctgtac ttccaatcca
     1861 atgcaatgga taagaaatac tcaataggct tagatatcgg cacaaatagc gtcggatggg
     1921 cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt caaggttctg ggaaatacag
     1981 accgccacag tatcaaaaaa aatcttatag gggctctttt atttgacagt ggagagacag
     2041 cggaagcgac tcgtctcaaa cggacagctc gtagaaggta tacacgtcgg aagaatcgta
     2101 tttgttatct acaggagatt ttttcaaatg agatggcgaa agtagatgat agtttctttc
     2161 atcgacttga agagtctttt ttggtggaag aagacaagaa gcatgaacgt catcctattt
     2221 ttggaaatat agtagatgaa gttgcttatc atgagaaata tccaactatc tatcatctgc
     2281 gaaaaaaatt ggtagattct actgataaag cggatttgcg cttaatctat ttggccttag
     2341 cgcatatgat taagtttcgt ggtcattttt tgattgaggg agatttaaat cctgataata
     2401 gtgatgtgga caaactattt atccagttgg tacaaaccta caatcaatta tttgaagaaa
     2461 accctattaa cgcaagtgga gtagatgcta aagcgattct ttctgcacga ttgagtaaat
     2521 caagacgatt agaaaatctc attgctcagc tccccggtga gaagaaaaat ggcttatttg
     2581 ggaatctcat tgctttgtca ttgggtttga cccctaattt taaatcaaat tttgatttgg
     2641 cagaagatgc taaattacag ctttcaaaag atacttacga tgatgattta gataatttat
     2701 tggcgcaaat tggagatcaa tatgctgatt tgtttttggc agctaagaat ttatcagatg
     2761 ctattttact ttcagatatc ctaagagtaa atactgaaat aactaaggct cccctatcag
     2821 cttcaatgat taaacgctac gatgaacatc atcaagactt gactctttta aaagctttag
     2881 ttcgacaaca acttccagaa aagtataaag aaatcttttt tgatcaatca aaaaacggat
     2941 atgcaggtta tattgatggg ggagctagcc aagaagaatt ttataaattt atcaaaccaa
     3001 ttttagaaaa aatggatggt actgaggaat tattggtgaa actaaatcgt gaagatttgc
     3061 tgcgcaagca acggaccttt gacaacggct ctattcccca tcaaattcac ttgggtgagc
     3121 tgcatgctat tttgagaaga caagaagact tttatccatt tttaaaagac aatcgtgaga
     3181 agattgaaaa aatcttgact tttcgaattc cttattatgt tggtccattg gcgcgtggca
     3241 atagtcgttt tgcatggatg actcggaagt ctgaagaaac aattacccca tggaattttg
     3301 aagaagttgt cgataaaggt gcttcagctc aatcatttat tgaacgcatg acagcgtttg
     3361 ataaaaatct tccaaatgaa aaagtactac caaaacatag tttgctttat gagtatttta
     3421 cggtttataa cgaattgaca aaggtcaaat atgttactga aggaatgcga aaaccagcat
     3481 ttctttcagg tgaacagaag aaagccattg ttgatttact cttcaaaaca aatcgaaaag
     3541 taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat agaatgtttt gatagtgttg
     3601 aaatttcagg agttgaagat agatttaatg cttcattagg tacctaccat gatttgctaa
     3661 aaattattaa agataaagat tttttggata atgaagaaaa tgaagatatc ttagaggata
     3721 ttgttttaac attgacctta tttgaagata gggagatgat tgaggaaaga cttaaaacat
     3781 atgctcacct ctttgatgat aaggtgatga aacagcttaa acgtcgccgt tatactggtt
     3841 ggggagcgtt gtctcgaaaa ttgattaatg gtattaggga taagcaatct ggcaaaacaa
     3901 tattagattt tttgaaatca gatggttttg ccaatcgcaa ttttatggcc ctgatccatg
     3961 atgatagttt gacatttaaa gaagacattc aaaaagcaca agtgtctgga caaggcgata
     4021 gtttacatga acatattgca aatttagctg gtagccctgc tattaaaaaa ggtattttac
     4081 agactgtaaa agttgttgat gaattggtca aagtaatggg gcggcataag ccagaaaata
     4141 tcgttattga aatggcacgt gaaaatcaga caactcaaaa gggccagaaa aattcgcgag
     4201 agcgtatgaa acgaatcgaa gaaggtatca aagaattagg aagtcagatt cttaaagagc
     4261 atcctgttga aaatactcaa ttgcaaaatg aaaagctcta tctctattat ctccaaaatg
     4321 gaagagacat gtatgtggac caagaattag atattaatcg tttaagtgat tatgatgtcg
     4381 atcacattgt tccacaaagt ttccttgccg acgattcaat agacaataag gtcttaacgc
     4441 gttctgataa aaatcgtggt aaatcggata acgttccaag tgaagaagta gtcaaaaaga
     4501 tgaaaaacta ttggagacaa cttctaaacg ccaagttaat cactcaacgt aagtttgata
     4561 atttaacgaa agctgaacgt ggaggtttga gtgaacttga taaagctggt tttatcaaac
     4621 gccaattggt tgaaactcgc gccatcacta agcatgtggc acaaattttg gatagtcgca
     4681 tgaatactaa atacgatgaa aatgataaac ttattcgaga ggttaaagtg attaccttaa
     4741 aatctaaatt agtttctgac ttccgaaaag atttccaatt ctataaagta cgtgagatta
     4801 acaattacca tcatgcccat gatgcgtatc taaatgccgt cgttggaact gctttgatta
     4861 agaaatatcc agcgcttgaa tcggagtttg tctatggtga ttataaagtt tatgatgttc
     4921 gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt
     4981 actctaatat catgaacttc ttcaaaacag aaattacact tgcaaatgga gagattcgca
     5041 aagcccctct aatcgaaact aatggggaaa ctggagaaat tgtctgggat aaagggcgag
     5101 attttgccac agtgcgcaaa gtattgtcca tgccccaagt caatattgtc aagaaaacag
     5161 aagtacagac aggcggattc tccaaggagt caattttacc aaaaagaaat tcggacaagc
     5221 ttattgctcg taaaaaagac tgggatccaa aaaaatatgg tggttttgat agtccaacgg
     5281 tagcttattc agtcctagtg gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat
     5341 ccgttaaaga gttactaggg atcacaatta tggaaagaag ttcctttgaa aaaaatccga
     5401 ttgacttttt agaagctaaa ggatataagg aagttaaaaa agacttaatc attaaactac
     5461 ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg gatgctggct agtgccggag
     5521 aattacaaaa aggaaatgag ctggctctgc caagcaaata tgtgaatttt ttatatttag
     5581 ctagtcatta tgaaaagttg aagggtagtc cagaagataa cgaacaaaaa caattgtttg
     5641 tggagcagca taagcattat ttagatgaga ttattgagca aatcagtgaa ttttctaagc
     5701 gtgttatttt agcagatgcc aatttagata aagttcttag tgcatataac aaacatagag
     5761 acaaaccaat acgtgaacaa gcagaaaata ttattcattt atttacgttg acgaatcttg
     5821 gagctcccgc tgcttttaaa tattttgata caacaattga tcgtaaacga tatacgtcta
     5881 caaaagaagt tttagatgcc actcttatcc atcaatccat cactggtctt tatgaaacac
     5941 gcattgattt gagtcagcta ggaggtgact aataacattg gaagtggata acggatccgc
     6001 gatcgcggcg cgccacctgg tggccggccg gtaccacgcg tgcgcgctga tccggctgct
     6061 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     6121 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     6181 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
     6241 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
     6301 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
     6361 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     6421 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     6481 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     6541 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     6601 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6661 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6721 tgagacaata accctgataa atgcttcaat aacattgaaa aaggaagagt atgagtattc
     6781 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6841 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6901 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6961 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     7021 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7081 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7141 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7201 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7261 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     7321 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7381 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7441 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7501 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7561 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7621 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7681 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7741 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7801 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7861 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7921 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7981 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8041 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8101 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8161 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8221 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8281 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8341 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8401 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8461 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8521 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     8581 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     8641 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     8701 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     8761 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     8821 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     8881 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     8941 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     9001 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     9061 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     9121 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     9181 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     9241 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     9301 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     9361 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     9421 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     9481 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     9541 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     9601 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     9661 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     9721 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     9781 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     9841 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     9901 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     9961 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
    10021 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccg
//