LOCUS       sequence-486055-       10951 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Edited bRA66 containing gRNA to create S. cerevisiae deltaMic60 and 
            S. cerevisiae Mic60(1-317).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10951)
  AUTHORS   Benning FMC, Bell TA, Nguyen TH, Syau D, Connell LB, Liao YT, 
            Keating MP, Coughlin M, Nordstrom AEH, Ericsson M, daCosta CJB, Chao
            LH
  TITLE     Ancestral sequence reconstruction of the Mic60 Mitofilin domain 
            reveals residues supporting respiration in yeast.
  JOURNAL   Protein Sci. 2025 Jul;34(7):e70207. doi: 10.1002/pro.70207.
  PUBMED    40545685
REFERENCE   2  (bases 1 to 10951)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2025-07"; volume: "34(7):e70207. doi"; pages: " 
            10.1002/pro.70207"
COMMENT     Sequence Label: bRA66-deltaMic60
FEATURES             Location/Qualifiers
     source          1..10951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            complement(284..1859)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(284..481)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(487..1515)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(487..1515)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(1516..1859)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        1993..2434
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1993..2110
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2405..2428
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2454..6557
                     /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             2454..6557
                     /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             6570..6590
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6594..6614
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6618..6638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6639..6704
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      6723..6970
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6739..6757)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(7030..7485)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7167..7188)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7379..7398
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7611..7633
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7625..7642
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7626..7642
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7649..7668
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7649..7667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7700..7716
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7700..7716
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        7748..8016
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        8037..8111
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      8116..8135
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(8178..8196)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8217..8233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8217..8233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8230..8252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8241..8257
                     /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)."
     promoter        complement(join(8265..8271,8272..8289,8290..8295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8310..8331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8448..8465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8619..9207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8699..8718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9378..10169,10170..10238))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10001..10020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10239..10343)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    10380..10883
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     10924..10942
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcgaattcg
      241 agctcgttta aactggatgg cggcgttagt atcgaatcga cagcagtata gcgaccagca
      301 ttcacatacg attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga
      361 tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta
      421 caatataaaa aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag
      481 tactgattat tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
      541 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
      601 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
      661 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
      721 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
      781 ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga
      841 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag
      901 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc
      961 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac
     1021 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt
     1081 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc
     1141 cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc
     1201 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc
     1261 gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg
     1321 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc
     1381 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag
     1441 gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc
     1501 aggcttttta cccatggttg tttatgttcg gatgtgatgt gagaactgta tcctagcaag
     1561 attttaaaag gaagtatatg aaagaagaac ctcagtggca aatcctaacc ttttatattt
     1621 ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc gtttccctgc
     1681 tcgcaggtcc gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt
     1741 atggatgcaa atgattatac atggggatgt atgggctaaa tgtacgggcg acagtcacat
     1801 catgcccctg agctgcgcac gtcaagactg tcaaggaggg tattctgggc ctccatgtcg
     1861 ctggccgggt gacccggcgg ggacgaggca agctaaacag atctgtaaag agccccatta
     1921 tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc
     1981 tatattgaag tacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct
     2041 cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg
     2101 cactgctccg aacaataaag attctacaat actagctttt atggttatga agaggaaaaa
     2161 ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga
     2221 tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat
     2281 ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt
     2341 tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat
     2401 tgttaatata cctctatact ttaacgtcaa ggagaaaaaa cccggatctc aaaatggata
     2461 aaaagtatag tattggttta gatattggta ctaactctgt gggttgggca gttatcactg
     2521 acgaatataa agttccatca aagaaattta aggtgttagg taacactgac agacactcaa
     2581 taaaaaagaa tcttatcggt gctcttttgt tcgactccgg tgaaactgcc gaggctacac
     2641 gtttaaaaag aacagcaaga agaagatata cccgtagaaa aaatagaata tgttatttac
     2701 aagaaatctt ttctaatgaa atggctaaag ttgatgattc ctttttccat agattggaag
     2761 agtcattttt ggttgaagaa gacaaaaagc atgagagaca tccaatcttt gggaatatag
     2821 ttgatgaagt ggcttaccat gaaaaatatc ctaccattta tcatttaaga aagaaattgg
     2881 tagattcaac tgataaagct gaccttagat taatctattt agcacttgcc catatgatta
     2941 aatttagagg tcattttttg attgaaggtg atttgaaccc agataattct gacgtggata
     3001 aattatttat tcaattagtc caaacctaca accaattatt tgaggaaaat ccaattaatg
     3061 ctagtggtgt cgatgccaaa gctatattat cagccagatt atcaaaatct agacgtttgg
     3121 aaaatttgat tgcccaattg ccaggagaaa aaaagaatgg attatttgga aacttgatcg
     3181 cattatcatt gggtttgaca ccaaatttta aatctaattt tgatttagct gaagatgcta
     3241 aattacaatt atcaaaagac acctatgacg acgatttgga caatttactt gctcaaattg
     3301 gtgatcaata tgcagatttg ttcttagctg ctaaaaactt atctgatgct attttgttgt
     3361 ctgatatttt gagagtgaac acagaaataa ccaaagctcc attatcagca tctatgatca
     3421 aacgttatga tgaacaccat caggatttga ctttattgaa agctttggtg agacaacaat
     3481 tgccagagaa gtataaagaa atctttttcg atcaatctaa aaacgggtat gcaggttata
     3541 ttgatggggg tgcctcccaa gaggaatttt acaaatttat aaaacctatt ttagaaaaga
     3601 tggatgggac tgaggaactt ttggtcaaat tgaacagaga agatttgtta cgtaaacaga
     3661 gaacttttga taatggtagt atacctcacc aaattcattt gggtgagttg catgcaattt
     3721 taagaagaca agaagatttt tatccatttt taaaagataa tagagaaaaa atcgagaaaa
     3781 ttttaacctt tagaattcca tactatgttg ggcctttggc tagaggtaat tcaagatttg
     3841 cctggatgac acgtaaatca gaagaaacta ttaccccttg gaattttgaa gaggttgttg
     3901 ataaaggagc atcagcacag agttttattg aaagaatgac caatttcgat aaaaacttac
     3961 caaatgaaaa agttttacca aaacattcct tgttatacga atattttact gtttacaatg
     4021 aacttacaaa ggttaaatat gttactgaag gtatgcgtaa gccagccttt ttatctggag
     4081 aacagaaaaa ggcaatagtt gatttattgt ttaaaacaaa tagaaaagtt actgttaaac
     4141 aattaaaaga agattacttt aagaaaattg aatgttttga ttcagttgaa atcagtggtg
     4201 ttgaagacag atttaatgct agtttaggaa cttaccatga tttacttaaa attatcaaag
     4261 ataaagattt cttggataac gaagaaaatg aagacatttt agaagacatt gttttaacct
     4321 taactttatt cgaagataga gagatgattg aagaacgttt gaagacttat gcacatttgt
     4381 ttgacgataa agtgatgaaa cagttgaaaa gaagacgtta tactggatgg ggtagattgt
     4441 ctcgtaaatt gatcaatgga attagagata aacaaagtgg taaaactatc ttggactttt
     4501 tgaaatctga cggatttgct aatagaaatt tcatgcaatt gatccacgac gatagtttga
     4561 catttaaaga agacatccaa aaggcccaag tgagtgggca aggtgattca ttacatgaac
     4621 atattgcaaa tttagccgga tctcctgcta ttaagaaagg gatattacaa actgttaaag
     4681 ttgtggatga attagtgaaa gtaatgggaa gacataaacc tgaaaacatt gtcattgaga
     4741 tggcaagaga aaatcaaact acacaaaaag gacagaaaaa tagtagagaa cgtatgaaaa
     4801 gaatagaaga gggtattaaa gaattgggta gtcaaatatt gaaagaacac ccagtggaaa
     4861 atacccagtt gcaaaatgaa aaattatatc tttactacct tcaaaatgga cgtgatatgt
     4921 atgttgatca ggaattagat ataaatagac tttcagatta tgatgtagat catatagttc
     4981 cacaatcttt cttgaaagat gattccatag acaataaagt attaactaga agtgataaaa
     5041 atagaggtaa aagtgataat gtcccaagtg aggaagtcgt caaaaagatg aaaaattact
     5101 ggcgtcaact tttgaatgct aaattaatta ctcaaagaaa atttgataat ttgactaaag
     5161 cagaaagagg tgggctttct gaattagata aagccgggtt cattaaaaga caattggtcg
     5221 aaactagaca aattactaaa catgttgccc aaattttaga ttcccgtatg aacactaagt
     5281 atgacgaaaa tgataagtta atacgtgagg ttaaagtcat tactttaaaa tcaaaacttg
     5341 tctctgattt cagaaaggat ttccaattct ataaagttag agaaattaat aattatcatc
     5401 atgctcatga tgcatatttg aatgctgtag ttggaactgc tttaatcaag aaatacccta
     5461 aattagaatc tgaatttgta tatggtgatt acaaagtcta tgatgttaga aagatgattg
     5521 ctaaatcaga acaagaaatt ggtaaagcta cagctaaata cttcttttac tctaacatta
     5581 tgaatttctt taaaacagaa attactttgg caaacggtga aattagaaaa agacctctta
     5641 ttgaaacaaa tggtgagact ggagagatag tttgggacaa agggcgtgat ttcgctactg
     5701 ttagaaaagt tttatcaatg ccacaagtta acattgtaaa gaaaacagag gttcaaactg
     5761 gtggtttctc aaaagaaagt attttgccta aaagaaatag tgataaattg attgccagaa
     5821 aaaaggattg ggatccaaag aaatatggtg gtttcgactc accaaccgta gcctattctg
     5881 ttttggttgt ggcaaaggtt gaaaagggta aaagtaaaaa gcttaaatca gtaaaagaac
     5941 ttttgggtat tacaataatg gaaagaagtt cctttgaaaa gaaccctatt gattttttgg
     6001 aagctaaagg ttataaggaa gtaaagaagg acttaataat caaattgcct aaatattctt
     6061 tatttgaatt agaaaatggg agaaaaagaa tgttggcttc tgctggagaa ttgcaaaagg
     6121 gtaatgaatt agcattgcct tccaaatatg ttaacttctt gtatttagct tcacactatg
     6181 aaaagttgaa agggtcacca gaagataacg agcaaaaaca attatttgtt gaacaacaca
     6241 aacactactt agatgagatt atagaacaaa ttagtgaatt cagtaaaaga gtgatattag
     6301 ctgatgcaaa tttagataaa gttttgtcag cctataacaa acatagagat aagccaatta
     6361 gagaacaagc agaaaacatt attcacttat ttacccttac caatttagga gcacctgctg
     6421 ctttcaagta ttttgataca acaattgatc gtaaaagata tacctcaaca aaagaagtct
     6481 tagacgccac cttaattcat caatcaatca ctggattgta tgagacaaga attgatttgt
     6541 ctcaattggg tggtgatgaa ggggctgatc ctaagaagaa aagaaaagtt gatccaaaga
     6601 aaaagcgtaa ggtggatcct aagaaaaaga gaaaggttga ctacaaagac catgacggtg
     6661 attataaaga tcatgacatc gactacaagg atgacgatga caagtgataa tctcttctcg
     6721 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     6781 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     6841 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     6901 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     6961 tttaatttgc ggccggtact gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     7021 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     7081 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     7141 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     7201 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     7261 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     7321 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     7381 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     7441 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     7501 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     7561 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     7621 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattggagct
     7681 ccaccgcggt ggcggccgct ctagaactag tggatccccc gggctgcagg aattcgatat
     7741 caagctttct ttgaaaagat aatgtatgat tatgctttca ctcatattta tacagaaact
     7801 tgatgttttc tttcgagtat atacaaggtg attacatgta cgtttgaagt acaactctag
     7861 attttgtagt gccctcttgg gctagcggta aaggtgcgca ttttttcaca ccctacaatg
     7921 ttctgttcaa aagattttgg tcaaacgctg tagaagtgaa agttggtgcg catgtttcgg
     7981 cgttcgaaac ttctccgcag tgaaagataa atgatcagat tcttgactgt gaaatagttt
     8041 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     8101 ccgagtcggt ggtgcttttt ttgtttttta tgtctaccgt cgacctcgag ggggggcccg
     8161 gtacccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     8221 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     8281 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     8341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     8401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     8461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     8521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     8581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     8641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     8701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     8821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     9001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     9061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     9121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     9181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     9241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     9301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     9361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     9421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     9481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9661 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9961 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    10021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    10081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    10141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    10201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    10261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    10321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtccttttca
    10381 tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa
    10441 tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga
    10501 ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct ctcaggtatt
    10561 aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt
    10621 acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa
    10681 atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc
    10741 ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata
    10801 aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc
    10861 gtgtaagtta caggcaagcg atccgtctaa gaaaccatta ttatcatgac attaacctat
    10921 aaaaataggc gtatcacgag gccctttcgt c
//
