LOCUS       sequence-217190-        9761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and purification of NLSH2BCas9 for fungal gene editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9761)
  AUTHORS   Wang Q, Cobine PA, Coleman JJ
  TITLE     Efficient genome editing in Fusarium oxysporum based on CRISPR/Cas9 
            ribonucleoprotein complexes.
  JOURNAL   Fungal Genet Biol. 2018 May 12. pii: S1087-1845(18)30081-1. doi: 
            10.1016/j.fgb.2018.05.003.
  PUBMED    29763675
REFERENCE   2  (bases 1 to 9761)
  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: "Fungal 
            Genet Biol. 2018 May 12. pii: S1087-1845(18)30081-1. doi: 
            10.1016/j.fgb.2018.05.003."
COMMENT     Sequence Label: pHis-parallel1-NLSH2BCas9
FEATURES             Location/Qualifiers
     source          1..9761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..42
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             223..4326
                     /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
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             223..4326
                     /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
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4330..4350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4375..4392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4445..4463)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4459..4506
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4543..4998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4630..4649)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4840..4861
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5024..5128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5129..5197,5198..5989)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5347..5366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6160..6748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6649..6668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6902..6919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6934..7073
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6999..7018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7159..7181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7175..7366)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(8175..9257)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     9219..9238
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(9258..9335)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     9541..9560
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     9644..9663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9644..9662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    9663..9687
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             9702..9724
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             9744..9761
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gattacgata tcccaacgac cgaaaacctg tattttcagg gcgccatgga tccgatggct
       61 cccaaggctg ctgacaagaa gcccgcctcc aaggctcccg ctactgcctc caaggctcct
      121 gagaagaagg atgctggcaa gaagactgcc gcctctggtg acaagaagaa gcgctccaag
      181 acccgcaagg agacctactc ttcttacatc tacaaggtcg acatggacaa gaagtatagc
      241 atcgggctgg acattggaac gaactcggtt ggttgggctg tgattacgga cgaatacaag
      301 gtgccatcca agaagtttaa ggtcctggga aacaccgacc gtcactcaat caagaagaat
      361 ctcattggag ccctgctctt cgatagtggg gagaccgccg aagctactcg actgaagcga
      421 acggctcgcc ggcgttatac acgacgcaag aatcgcatct gctacctcca ggagattttc
      481 agcaacgaaa tggctaaggt tgatgactca ttctttcatc gactcgaaga aagtttcttg
      541 gtcgaggagg ataagaagca cgagcgccat ccgatctttg gtaacattgt ggatgaggtt
      601 gcctatcacg aaaagtaccc aactatctat catcttcgta agaagctggt cgatagcacg
      661 gacaaggctg atttgcgact tatctacctg gcactcgcgc acatgattaa gttccgcggc
      721 cattttctta tcgagggtga cctgaacccc gataattctg acgttgataa gctcttcatc
      781 cagttggtcc aaacctacaa tcagctgttt gaggaaaacc ctattaatgc atctggcgtg
      841 gacgccaagg ctatcctttc ggcgcgcctg tctaagtcgc ggcgtttgga gaaccttatc
      901 gcacaactcc ccggcgaaaa gaagaacggc ctcttcggta atttgattgc gttgtcactt
      961 ggtctgactc ctaacttcaa gagtaatttt gacctggcag aggatgcgaa gctccagttg
     1021 tctaaggata cgtatgatga cgatctcgac aacttgcttg cccaaatcgg tgaccagtac
     1081 gctgatcttt tcctggccgc taagaatctc tcagatgcaa tcctgctcag tgacattttg
     1141 cgggtcaaca ccgagattac taaggccccc ctgtcagcta gtatgatcaa gcggtatgat
     1201 gagcaccatc aggacctcac cttgcttaag gccctcgtgc gtcagcaatt gcctgagaag
     1261 tataaggaaa tcttctttga ccaatccaag aacggatacg cagggtatat tgatggcggt
     1321 gcgagccagg aggaattcta caagtttatc aagccgattt tggagaagat ggacggcact
     1381 gaggaactgc tcgtcaagct gaatcgcgaa gatttgcttc gtaagcaacg aacgttcgac
     1441 aacggctcca tcccgcacca gattcatctg ggcgagctcc acgccatcct tcgacgccag
     1501 gaagatttct acccatttct gaaggacaac cgtgagaaga tcgaaaagat tcttacattc
     1561 cgaatcccct actatgtggg acctttggcc cgtgggaatt cccgatttgc ttggatgacc
     1621 cgaaagagcg aggaaaccat cactccgtgg aacttcgagg aagtcgtgga caagggtgca
     1681 tccgcgcaga gcttcattga gcggatgacc aattttgata agaaccttcc gaatgaaaag
     1741 gtcctgccaa agcattcgct gctctacgag tatttcaccg tgtataacga actgactaag
     1801 gtcaagtacg tgacggaggg aatgcggaag ccagccttcc tctcagggga acaaaagaag
     1861 gctatcgtcg atttgctttt taagaccaat cgtaaagtga ctgttaagca gctgaaggag
     1921 gattatttca agaagattga atgtttcgac tccgtcgaga tcagcggcgt ggaagatcgc
     1981 tttaacgctt ccctcggtac ctaccacgac ctgctcaaga tcattaagga caaggatttc
     2041 ctcgataacg aggaaaatga ggacatcttg gaagatattg tcctcacgtt gacacttttt
     2101 gaggaccgcg aaatgatcga ggaacggctc aagacatatg cccatttgtt cgacgataag
     2161 gtgatgaagc agctgaagcg gcgtcgatac accggatggg gtcgccttag ccggaagctg
     2221 atcaacggca ttcgagataa gcaatctggt aagactatct tggatttcct taagtcggac
     2281 ggcttcgcca accgcaattt tatgcagctt attcacgacg attccctgac gttcaaggag
     2341 gacatccaga aggcacaagt ctcaggacaa ggggattccc tgcacgagca tatcgccaac
     2401 ctggctggat ccccggcgat caagaagggg attcttcaga ccgtcaaggt tgtcgacgag
     2461 ctggtcaagg tgatgggccg tcataagcca gaaaacatcg tgattgagat ggcccgagaa
     2521 aatcagacca ctcaaaaggg tcagaagaac agccgcgagc ggatgaagcg gatcgaggaa
     2581 ggcattaagg aacttggttc tcagatcctg aaggagcacc ctgttgaaaa cacacagctc
     2641 caaaatgaga agctgtatct ctactatttg caaaatggac gcgacatgta cgtcgatcag
     2701 gagctcgaca ttaaccggtt gtcggactac gatgttgacc atatcgtccc gcaatccttc
     2761 cttaaggacg atagcattga taacaaggtg ctgactcgct cagataagaa ccggggcaag
     2821 tccgacaatg ttccaagcga ggaagtggtt aagaagatga agaactactg gcgccaattg
     2881 cttaatgcca agctcatcac acagcgcaag tttgacaact tgaccaaggc cgagcgggga
     2941 gggctgagtg aactcgataa ggctggcttc atcaagcgtc aactcgtgga gacgcgacag
     3001 atcacaaagc acgttgctca gattctggac tcccggatga acacaaagta cgacgagaat
     3061 gataagctca tccgtgaagt taaggtcatt accctcaagt ctaagttggt gtcggatttc
     3121 cgcaaggact tccaatttta taaggttcgg gagatcaaca attatcacca tgcacatgat
     3181 gcgtacctca acgcagtcgt gggaactgcg ctcatcaaga agtatcccaa gttggagtcc
     3241 gaattcgtct acggggatta taaggtttac gacgtccgca agatgatcgc caagagtgag
     3301 caggaaattg gcaaggccac ggctaagtat ttcttttact ccaacatcat gaatttcttt
     3361 aagacggaga tcacactcgc caatggagaa atccgtaagc gacctttgat tgagaccaac
     3421 ggcgagactg gtgaaatcgt ttgggataag gggcgcgact tcgctaccgt gcggaaggtt
     3481 ctgagcatgc cgcaagtcaa tatcgtcaag aaaaccgagg tgcagacagg cggtttctct
     3541 aaggaatcga ttcttccaaa gcgtaactct gacaagctga tcgctcgaaa gaaggattgg
     3601 gaccccaaga agtatggagg gttcgattct cctacagtgg catactcggt tctcgttgtc
     3661 gcgaaggttg agaagggaaa gtctaagaag ctgaagtcgg tcaaggaact gctcgggatc
     3721 accattatgg agcgctccag cttcgaaaag aatcccatcg actttctcga ggccaagggc
     3781 tataaggaag tcaagaagga tcttatcatt aagctgccta agtactcttt gttcgagctt
     3841 gaaaacggtc gaaagcgaat gctcgcatcg gcaggagagt tgcagaaggg gaatgaattg
     3901 gcacttccct caaagtacgt gaacttcctg tatctcgcgt cccactacga gaagctgaag
     3961 ggtagccctg aggacaacga acagaagcaa ctttttgttg agcaacacaa gcattatctg
     4021 gatgagatca ttgaacagat ttcagagttc agtaagcgcg tcatcctcgc cgatgctaat
     4081 ctcgacaagg tgttgtcggc ctacaacaag caccgtgaca agccgatccg agagcaggct
     4141 gaaaatatca ttcatctgtt caccctcact aacttgggag caccagcagc gttcaagtat
     4201 tttgatacga caatcgaccg taagcgatac acgtccacaa aggaggtgct tgatgcgacc
     4261 ctgattcatc aatccatcac tgggctctat gaaacccgta tcgaccttag tcaactgggg
     4321 ggcgaccctc ccaagaagaa gcgcaaggtc tgaaagcttg cggccgcact cgagcaccac
     4381 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     4441 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     4501 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     4561 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4621 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4681 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4741 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4801 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4861 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4921 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4981 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5041 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5101 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5161 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5221 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5281 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5341 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5401 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5461 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5521 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5581 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5641 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     5701 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5761 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5821 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5881 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5941 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6001 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6301 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6781 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6841 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6901 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     6961 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     7021 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     7081 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7141 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7201 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     7261 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     7321 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     7381 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     7441 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     7501 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     7561 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     7621 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     7681 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     7741 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     7801 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     7861 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     7921 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     7981 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     8041 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     8101 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     8161 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     8221 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     8281 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     8341 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     8401 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     8461 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     8521 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     8581 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     8641 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     8701 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     8761 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     8821 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     8881 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     8941 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     9001 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     9061 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     9121 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     9181 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     9241 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     9301 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     9361 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     9421 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     9481 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     9541 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     9601 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     9661 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     9721 gagatataca tatgtcgtac taccatcacc atcaccatca c
//
