LOCUS       sequence-367190-       11220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LexA-HBD-B112 under the control of pAct1 and Cas9 under 
            the control of LexA-HBD-B112+Beta-estradiol inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11220)
  AUTHORS   Azizoglu A, Loureiro C, Venetz J, Brent R
  TITLE     Autorepression-Based Conditional Gene Expression System in Yeast for
            Variation-Suppressed Control of Protein Dosage.
  JOURNAL   Curr Protoc. 2023 Jan;3(1):e647. doi: 10.1002/cpz1.647.
  PUBMED    36708363
REFERENCE   2  (bases 1 to 11220)
  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: "Curr 
            Protoc."; date: "2023-01"; volume: "3(1):e647. doi"; pages: " 
            10.1002/cpz1.647"
COMMENT     Sequence Label: P2061
FEATURES             Location/Qualifiers
     source          1..11220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..55)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(37..55)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        276..351
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(951..1202,1203..1556)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     1494..1515
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             1575..2510
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             1623..2513
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        join(3278..3283,3284..3299,3300..3305)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3314..3330
                     /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)."
     CDS             3351..4166
                     /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             3351..4166
                     /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     complement(3424..3443)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4172..4369
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      4413..5755
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5376..5423)
                     /label=FRT (mutant)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT site with a mutation in the core sequence (Azam 
                     et al., 1997)"
     protein_bind    complement(5376..5423)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      5805..6393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6294..6313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6547..6564
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6649..6665
                     /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)."
     primer_bind     6673..6689
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6673..6689
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6710..6728
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(6819..6839)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6846..10949)
                     /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             complement(6846..10949)
                     /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"
ORIGIN
        1 tacaggttta tatattattc tgtttctcca attcgcccta tagtgagtcg tattaaccgg
       61 tgtatactct ttcttcaaca attaaatact ctcggtagcc aagttggttt aaggcgcaag
      121 actgtaattt atcactacga aatcttgaga tcgggcgttc gactcgcccc cgggagagat
      181 ggccggcatg gtcccagcct cctcgctggc gccggctggg caacaccttc gggtggcgaa
      241 tgggaccgga gaccggccct taagtcctgg tctccgtttt agagctagaa atagcaagtt
      301 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgg
      361 cgcgcctctg tcgactctag agaagcgggt aagctgccac agcaattaat gcacaacatt
      421 taacctacat tcttccttat cggatcctca aaacccttaa aaacatatgc ctcaccctaa
      481 catattttcc aattaaccct caatatttct ctgtcacccg gcctctattt tccattttct
      541 tctttacccg ccacgcgttt ttttctttca aatttttttc ttccttcttc tttttcttcc
      601 acgtcctctt gcataaataa ataaaccgtt ttgaaaccaa actcgcctct ctctctcctt
      661 tttgaaatat ttttgggttt gtttgatcct ttccttccca atctctcttg tttaatatat
      721 attcatttat atcacgctct ctttttatct tccttttttt cctctctctt gtattcttcc
      781 ttcccctttc tactcaaacc aagaagaaaa agaaaaggtc aatctttgtt aaagaatagg
      841 atcttctact acatcagctt ttagattttt cacgcttact gcttttttct tcccaagatc
      901 gaaaatttac tgaattaaca actagtggat cccccgggct gcaggaattc atgaaagcgt
      961 taacggccag gcaacaagag gtgtttgatc tcatccgtga tcacatcagc cagacaggta
     1021 tgccgccgac gcgtgcggaa atcgcgcagc gtttggggtt ccgttcccca aacgcggctg
     1081 aagaacatct gaaggcgctg gcacgcaaag gcgttattga aattgtttcc ggcgcatcac
     1141 gcgggattcg tctgttgcag gaagaggaag aagggttgcc gctggtaggt cgtgtggctg
     1201 ccggtgaacc acttctggcg caacagcata ttgaaggtca ttatcaggtc gatccttcct
     1261 tattcaagcc gaatgctgat ttcctgctgc gcgtcagcgg gatgtcgatg aaagacatcg
     1321 gcattatgga tggtgacttg ctggcagtgc ataaaactca ggatgtacgt aacggtcagg
     1381 tcgttgtcgc acgtattgat gacgaggtta ccgttaagcg cctgaaaaaa cagggcaata
     1441 aagtcgaact gttgccagaa aatagcgagt ttaaaccaat tgtcgtagat cttcgtcagc
     1501 agagcttcac cattgaaggg ctggcggttg gggttattcg caacggcgac tggctgggtt
     1561 ctaaggatat ctctgctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga
     1621 tcaaacgctc taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg
     1681 ccttgttgga tgctgagccc cccatactct attccgagta tgatcctacc agacccttca
     1741 gtgaagcttc gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga
     1801 tcaactgggc gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc
     1861 ttctagaatg tgcctggtta gagatcctga tgattggttt ggtctggcgc tccatggagc
     1921 acccagtgaa gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg
     1981 tagagggcat ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga
     2041 tgaatctgca gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag
     2101 tgtacacatt tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag
     2161 tcctggacaa gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc
     2221 agcagcagca ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga
     2281 gtaacaaagg catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg
     2341 acctgctgct ggagatgctg gacgcccacc gcctacatgc gcccactagc cgtggagggg
     2401 catccgtgga ggagacggac caaagccact tggccactgc gggctctact tcatcgcatt
     2461 ccttgcaaaa gtattacatc acgggggagg cagagggttt ccctgccaca gtcgatatca
     2521 tgcaggatct gccgggcaac gataacagca ccgcgggcga atttccaggt attactttga
     2581 gaatccaaga aactgatatg ttgtataaag gtgatacttt gtatttggat tggttggaag
     2641 atggtattgc tgaattggtt ttcgatgctc caggttctgt taataaattg gatactgctg
     2701 ttgcttcttt gggtgaagct attggtgttt tggaacaaca atctgatttg atttgggaaa
     2761 ctttgactgt taaagatgct aaagttaatt tcgattctgg tttggaaaaa ttcgaagaag
     2821 ctattccatc tgctgatgat ttcgatccag ttgctgaaag aagatcctct ggtgaattta
     2881 gagctgaaag acattctggt ggtactgatt tgtgtttcaa gctttaatta acgacatgga
     2941 ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg
     3001 tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca tacattttga
     3061 tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa
     3121 cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa
     3181 aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg
     3241 atacagttct cacatcacat ccgaacataa agagctgttg acaattaatc atcggctcgc
     3301 ataatctgtg gaattgtgag cggataacaa tttcacacag gaaacataaa atgagccata
     3361 ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat gctgatttat
     3421 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     3481 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     3541 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     3601 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
     3661 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     3721 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     3781 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     3841 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     3901 agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     3961 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     4021 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     4081 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     4141 atttgatgct cgatgagttt ttctaatcag tactgacaat aaaaagattc ttgttttcaa
     4201 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
     4261 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     4321 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgg gtacccagct
     4381 tcacatttcc ccgaaaagtg ccacctgacg tcaacgaagc atctgtgctt cattttgtag
     4441 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4501 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     4561 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     4621 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     4681 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     4741 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     4801 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     4861 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     4921 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     4981 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5041 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5101 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     5161 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     5221 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     5281 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     5341 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tttagagaat
     5401 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     5461 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     5521 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     5581 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     5641 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     5701 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatatt
     5761 aattaaacac acacatggta ataactgcag atcaaaggat cttcttgaga tccttttttt
     5821 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5881 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5941 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6001 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6061 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6121 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6181 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6241 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6301 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6361 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6421 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6481 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6541 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6601 cccgcgcgtt ggccgattca ttaatgcctg caggtgcatc agcagctgtt gtgagcggat
     6661 aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgaa attaaccctc
     6721 actaaaggga acaaaagctg ggtacaagtg acaatatata tatatatata ataacgacat
     6781 cgttatctga aatacggccg ttaagcagaa gcatcttcaa cttttctttt cttttttggt
     6841 ccaccgtcgc ctcccagctg agacaggtcg atccgtgtct cgtacaggcc ggtgatgctc
     6901 tggtggatca gggtggcgtc cagcacctct ttggtgctgg tgtacctctt ccggtcgatg
     6961 gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat tggtcagggt aaacaggtgg
     7021 atgatattct cggcctgctc tctgatgggc ttatcccggt gcttgttgta ggcggacagc
     7081 actttgtcca gattagcgtc ggccaggatc actctcttgg agaactcgct gatctgctcg
     7141 atgatctcgt ccaggtagtg cttgtgctgt tccacaaaca gctgtttctg ctcattatcc
     7201 tcgggggagc ccttcagctt ctcatagtgg ctggccaggt acaggaagtt cacatatttg
     7261 gagggcaggg ccagttcgtt tcccttctgc agttcgccgg cagaggccag cattctcttc
     7321 cggccgtttt ccagctcgaa cagggagtac ttaggcagct tgatgatcag gtcctttttc
     7381 acttctttgt agcccttggc ttccagaaag tcgatgggat tcttctcgaa gctgcttctt
     7441 tccatgatgg tgatccccag cagctctttc acactcttca gtttcttgga cttgcccttt
     7501 tccactttgg ccaccaccag cacagaatag gccacggtgg ggctgtcgaa gccgccgtac
     7561 ttcttagggt cccagtcctt ctttctggcg atcagcttat cgctgttcct cttgggcagg
     7621 atagactctt tgctgaagcc gcctgtctgc acctcggtct ttttcacgat attcacttgg
     7681 ggcatgctca gcactttccg cacggtggca aaatcccggc ccttatccca cacgatctcc
     7741 ccggtttcgc cgtttgtctc gatcagaggc cgcttccgga tctcgccgtt ggccagggta
     7801 atctcggtct tgaaaaagtt catgatgttg ctgtagaaga agtacttggc ggtagccttg
     7861 ccgatttcct gctcgctctt ggcgatcatc ttccgcacgt cgtacacctt gtagtcgccg
     7921 tacacgaact cgctttccag cttagggtac tttttgatca gggcggttcc cacgacggcg
     7981 ttcaggtagg cgtcgtgggc gtggtggtag ttgttgatct cgcgcacttt gtaaaactgg
     8041 aaatccttcc ggaaatcgga caccagcttg gacttcaggg tgatcacttt cacttcccgg
     8101 atcagcttgt cattctcgtc gtacttagtg ttcatccggg agtccaggat ctgtgccacg
     8161 tgctttgtga tctgccgggt ttccaccagc tgtctcttga tgaagccggc cttatccagt
     8221 tcgctcaggc cgcctctctc ggccttggtc agattgtcga actttctctg ggtaatcagc
     8281 ttggcgttca gcagctgccg ccagtagttc ttcatcttct tcacgacctc ttcggagggc
     8341 acgttgtcgc tcttgccccg gttcttgtcg cttctggtca gcaccttgtt gtcgatggag
     8401 tcgtccttca gaaagctctg aggcacgata tggtccacat cgtagtcgga cagccggttg
     8461 atgtccagtt cctggtccac gtacatatcc cgcccattct gcaggtagta caggtacagc
     8521 ttctcgttct gcagctgggt gttttccacg gggtgttctt tcaggatctg gctgcccagc
     8581 tctttgatgc cctcttcgat ccgcttcatt ctctcgcggc tgttcttctg tcccttctgg
     8641 gtggtctggt tctctctggc catttcgatc acgatgttct cgggcttgtg ccggcccatc
     8701 actttcacga gctcgtccac caccttcact gtctgcagga tgcccttctt aatggcgggg
     8761 ctgccggcca gattggcaat gtgctcgtgc aggctatcgc cctggccgga cacctgggct
     8821 ttctggatgt cctctttaaa ggtcaggctg tcgtcgtgga tcagctgcat gaagtttctg
     8881 ttggcgaagc cgtcggactt caggaaatcc aggattgtct tgccggactg cttgtcccgg
     8941 atgccgttga tcagcttccg gctcagcctg ccccagccgg tgtatctccg ccgcttcagc
     9001 tgcttcatca ctttgtcgtc gaacaggtgg gcataggttt tcagccgttc ctcgatcatc
     9061 tctctgtcct caaacagtgt cagggtcagc acgatatctt ccagaatgtc ctcgttttcc
     9121 tcattgtcca ggaagtcctt gtccttgata attttcagca gatcgtggta tgtgcccagg
     9181 gaggcgttga accgatcttc cacgccggag atttccacgg agtcgaagca ctcgattttc
     9241 ttgaagtagt cctctttcag ctgcttcacg gtcactttcc ggttggtctt gaacagcagg
     9301 tccacgatgg cctttttctg ctcgccgctc aggaaggcgg gctttctcat tccctcggtc
     9361 acgtatttca ctttggtcag ctcgttatac acggtgaagt actcgtacag caggctgtgc
     9421 ttgggcagca ccttctcgtt gggcaggttc ttatcgaagt tggtcatccg ctcgatgaag
     9481 ctctgggcgg aagcgccctt gtccaccact tcctcgaagt tccagggggt gatggtttcc
     9541 tcgctctttc tggtcatcca ggcgaatctg ctgtttcccc tggccagagg gcccacgtag
     9601 taggggatgc ggaaggtcag gatcttctcg atcttttccc ggttgtcctt caggaatggg
     9661 taaaaatctt cctgccgccg cagaatggcg tgcagctctc ccaggtggat ctggtggggg
     9721 atgctgccgt tgtcgaaggt ccgctgcttc cgcagcaggt cctctctgtt cagcttcacg
     9781 agcagttcct cggtgccgtc catcttttcc aggatgggct tgatgaactt gtagaactct
     9841 tcctggctgg ctccgccgtc aatgtagccg gcgtagccgt tcttgctctg gtcgaagaaa
     9901 atctctttgt acttctcagg cagctgctgc cgcacgagag ctttcagcag ggtcaggtcc
     9961 tggtggtgct cgtcgtatct cttgatcata gaggcgctca ggggggcctt ggtgatctcg
    10021 gtgttcactc tcaggatgtc gctcagcagg atggcgtcgg acaggttctt ggcggccaga
    10081 aacaggtcgg cgtactggtc gccgatctgg gccagcaggt tgtccaggtc gtcgtcgtag
    10141 gtgtccttgc tcagctgcag tttggcatcc tcggccaggt cgaagttgct cttgaagttg
    10201 ggggtcaggc ccaggctcag ggcaatcagg tttccgaaca ggccattctt cttctcgccg
    10261 ggcagctggg cgatcagatt ttccagccgt ctgctcttgc tcagtctggc agacaggatg
    10321 gccttggcgt ccacgccgct ggcgttgatg gggttttcct cgaacagctg gttgtaggtc
    10381 tgcaccagct ggatgaacag cttgtccacg tcgctgttgt cggggttcag gtcgccctcg
    10441 atcaggaagt ggccccggaa cttgatcatg tgggccaggg ccagatagat cagccgcagg
    10501 tcggccttgt cggtgctgtc caccagtttc tttctcaggt ggtagatggt ggggtacttc
    10561 tcgtggtagg ccacctcgtc cacgatgttg ccgaagatgg ggtgccgctc gtgcttctta
    10621 tcctcttcca ccaggaagga ctcttccagt ctgtggaaga agctgtcgtc caccttggcc
    10681 atctcgttgc tgaagatctc ttgcagatag cagatccggt tcttccgtct ggtgtatctt
    10741 cttctggcgg ttctcttcag ccgggtggcc tcggctgttt cgccgctgtc gaacagcagg
    10801 gctccgatca ggttcttctt gatgctgtgc cggtcggtgt tgcccagcac cttgaatttc
    10861 ttgctgggca ccttgtactc gtcggtgatc acggcccagc ccacagagtt ggtgccgatg
    10921 tccaggccga tgctgtactt cttgtccatg gggatccact agtatagaag tatagtaatt
    10981 tatgctgcaa aggtcctaat gtataaggaa agaagagaaa agaagaaaac aagagtttta
    11041 tatacataca gagcacatgc tctagaccct gggtgtatat acagttatgc tgtgagtata
    11101 tacagcacct agaccctggg tgtatataca gttatgctgt gagtatatac agcacctagg
    11161 gagctccaat tcgccctata ggcggccgcg aaaagaaata tagtcttcaa ggttatatta
//
