LOCUS       sequence-330060-       10771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NLS-AncBE4max-NLS with an N-terminal His Tag (6xHis) for 
            bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10771)
  AUTHORS   Zou RS, Liu Y, Wu B, Ha T
  TITLE     Cas9 deactivation with photocleavable guide RNAs.
  JOURNAL   Mol Cell. 2021 Apr 1;81(7):1553-1565.e8. doi: 
            10.1016/j.molcel.2021.02.007. Epub 2021 Mar 3.
  PUBMED    33662274
REFERENCE   2  (bases 1 to 10771)
  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: 
            "10.1016/j.molcel.2021.02.007"; journalName: "Mol Cell"; date: 
            "2021-04-1- 1"; volume: "81"; issue: "7"; pages: "1553-1565.e8"
COMMENT     Sequence Label: pET42b-AncBE4max
FEATURES             Location/Qualifiers
     source          1..10771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             118..138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(919..942,943..945,946..5019)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             919..5019
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5050..5298
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5329..5577
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5620..5640
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5706..5724)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5720..5767
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5804..6259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5891..5910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6101..6122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6351..7166)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6351..7166)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7074..7093
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7158..7177
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7288..7876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7777..7796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8030..8047
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8062..8201
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8127..8146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8287..8309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8303..8494)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     YELYRSCLARFGDDGENL"
     CDS             complement(9303..10385)
                     /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="MKPVTLYDVAEYAGVSYQTVSRVVNHASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQLLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESWQ"
     primer_bind     10347..10366
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10386..10463)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     10669..10688
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagtc atcatcatca ccatcaccca
      121 aagaagaagc ggaaagtcag cagtgaaacc ggaccagtgg cagtggaccc aaccctgagg
      181 agacggattg agccccatga atttgaagtg ttctttgacc caagggagct gaggaaggag
      241 acatgcctgc tgtacgagat caagtggggc acaagccaca agatctggcg ccacagctcc
      301 aagaacacca caaagcacgt ggaagtgaat ttcatcgaga agtttacctc cgagcggcac
      361 ttctgcccct ctaccagctg ttccatcaca tggtttctgt cttggagccc ttgcggcgag
      421 tgttccaagg ccatcaccga gttcctgtct cagcacccta acgtgaccct ggtcatctac
      481 gtggcccggc tgtatcacca catggaccag cagaacaggc agggcctgcg cgatctggtg
      541 aattctggcg tgaccatcca gatcatgaca gccccagagt acgactattg ctggcggaac
      601 ttcgtgaatt atccacctgg caaggaggca cactggccaa gatacccacc cctgtggatg
      661 aagctgtatg cactggagct gcacgcagga atcctgggcc tgcctccatg tctgaatatc
      721 ctgcggagaa agcagcccca gctgacattt ttcaccattg ctctgcagtc ttgtcactat
      781 cagcggctgc ctcctcatat tctgtgggct acaggcctga agtctggagg atctagcgga
      841 ggatcctctg gcagcgagac accaggaaca agcgagtcag caacaccaga gagcagtggc
      901 ggcagcagcg gcggcagcga caagaagtac agcatcggcc tggccatcgg caccaactct
      961 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     1021 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     1081 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     1141 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     1201 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     1261 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     1321 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     1381 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     1441 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     1501 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     1561 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     1621 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     1681 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     1741 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     1801 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     1861 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     1921 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     1981 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     2041 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     2101 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     2161 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     2221 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     2281 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     2341 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     2401 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     2461 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     2521 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     2581 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     2641 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     2701 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     2761 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     2821 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     2881 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     2941 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     3001 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     3061 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     3121 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     3181 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     3241 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     3301 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     3361 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     3421 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     3481 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     3541 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     3601 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     3661 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     3721 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     3781 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     3841 cgcgagatca acaactacca ccacgcccac gacgcctacc taaacgccgt cgtgggaacc
     3901 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     3961 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     4021 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     4081 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     4141 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     4201 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     4261 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     4321 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     4381 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     4441 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     4501 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     4561 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     4621 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     4681 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     4741 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     4801 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     4861 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     4921 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     4981 tacgagacac ggatcgacct gtctcagctg ggaggtgaca gcggcgggag cggcgggagc
     5041 ggggggagca ctaatctgag cgacatcatt gagaaggaga ctgggaaaca gctggtcatt
     5101 caggagtcca tcctgatgct gcctgaggag gtggaggaag tgatcggcaa caagccagag
     5161 tctgacatcc tggtgcacac cgcctacgac gagtccacag atgagaatgt gatgctgctg
     5221 acctctgacg cccccgagta taagccttgg gccctggtca tccaggattc taacggcgag
     5281 aataagatca agatgctgag cggaggatcc ggaggatctg gaggcagcac caacctgtct
     5341 gacatcatcg agaaggagac aggcaagcag ctggtcatcc aggagagcat cctgatgctg
     5401 cccgaagaag tcgaagaagt gatcggaaac aagcctgaga gcgatatcct ggtccatacc
     5461 gcctacgacg agagtaccga cgaaaatgtg atgctgctga catccgacgc cccagagtat
     5521 aagccctggg ctctggtcat ccaggattcc aacggagaga acaaaatcaa aatgctgtct
     5581 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc
     5641 taataattga ttaataccta ggctgctaaa caaagcccga aaggaagctg agttggctgc
     5701 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     5761 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     5821 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5881 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5941 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6001 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6061 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6121 ggaacaacac tcaaccatat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6181 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6241 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     6361 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     6421 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     6481 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     6541 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     6601 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     6661 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     6721 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     6781 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     6841 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     6901 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     6961 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     7021 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     7081 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     7141 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     7201 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7261 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgcta
     7321 attgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     7441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7501 tctgctaatc atgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7621 cacacagcca agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7681 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7861 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7921 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7981 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     8041 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     8101 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     8161 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     8221 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     8281 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     8341 agctgcggta aagctcataa gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     8401 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     8461 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     8521 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     8581 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     8641 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     8701 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     8761 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     8821 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     8881 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     8941 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     9001 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggagtgcaag attccgaata
     9061 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     9121 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     9181 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     9241 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     9301 gctcactgcc agctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     9361 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     9421 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     9481 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     9541 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     9601 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     9661 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     9721 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     9781 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     9841 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     9901 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     9961 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
    10021 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
    10081 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
    10141 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
    10201 aacaactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
    10261 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcatg gttcaccacg
    10321 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
    10381 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
    10441 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
    10501 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
    10561 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
    10621 gaaacaagcg ctcatgagcc caaagtggcg agcccgatct tccccatcgg tgatgtcggc
    10681 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
    10741 gtagaggatc gagatctcga tcccgcgaaa t
//
