LOCUS       sequence-467136-       10104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SpCAS9-Tyr-[gRNA: BsaI GFP dropout] with pan(OPT)ARS replication 
            origin and G418 selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10104)
  AUTHORS   Thornbury M, Knoops A, Summerby-Murray I, Dhaliwal J, Johnson S, 
            Utomo JC, Joshi J, Narcross L, Remondetto G, Pouliot M, Whiteway M, 
            Martin VJJ
  TITLE     Sequencing of a Dairy Isolate Unlocks Kluyveromyces marxianus as a 
            Host for Lactose Valorization.
  JOURNAL   ACS Synth Biol. 2025 Jul 8. doi: 10.1021/acssynbio.5c00096.
  PUBMED    40629255
REFERENCE   2  (bases 1 to 10104)
  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: "ACS Synth 
            Biol. 2025 Jul 8. doi: 10.1021/acssynbio.5c00096."
COMMENT     Sequence Label: pMTKM03
FEATURES             Location/Qualifiers
     source          1..10104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(129..717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(209..228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             1556..1567
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1574..2287
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2299..2370
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2386..2413
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        2427..2502
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     gene            2636..3992
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2636..2978
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2980..3789
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3047..3066)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3795..3992
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4143..4934,4935..5003))
                     /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     4766..4785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5004..5108)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5811..9914
                     /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             5811..9914
                     /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             9924..9944
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9960..9983
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      join(9987..10104,1..72)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
ORIGIN
        1 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
       61 ggacgctcga agtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      301 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      541 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      721 gatcctttga tcttttctac actagttcaa catctttgga taatatcaga atgagaaaga
      781 acagatacgc agtacgtttt ttggtgagct ctttgcactt ctttagttct ttccatcaat
      841 atcagttttt taaactttta ggactaaaag tgatgtttaa cttcaaaatg tttaaaattt
      901 tgttcttccc gacgttcatt aagaatacta atacacttta ataattattt taatattttg
      961 ttctaaataa tgacttttaa ttaaaaaaga taaaatataa aaacatcata ataactcacc
     1021 agaggttaag aacaaaaaaa caaattagat atctgctaat ccaatatagt taaatcaatc
     1081 tttccttggt ataatgggta tattacatat atttcaagga ccgacactcc taccaaatat
     1141 ctaaaattta ccatattaac ataacatgta tataaacgtc aaatcataat cagcactagt
     1201 atatgtgtta tgtagtatac tctttcttca acaattaaat actctcggta gccaagttgg
     1261 tttaaggcgc aagactgtaa tttatcacta cgaaatcttg agatcgggcg ttcgactcgc
     1321 ccccgggaga gatggccggc atggtcccag cctcctcgct ggcgccggct gggcaacacc
     1381 ttcgggtggc gaatgggact cgagaccgaa agtgaaacgt gatttcatgc gtcattttga
     1441 acattttgta aatcttattt aataatgtgt gcggcaattc acatttaatt tatgaatgtt
     1501 ttcttaacat cgcggcaact caagaaacgg caggttcgga tcttagctac tagagaaaga
     1561 ggagaaatac tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt
     1621 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga
     1681 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggttcc
     1741 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga
     1801 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg
     1861 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg
     1921 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagagg acggcaatat
     1981 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa
     2041 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt
     2101 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc
     2161 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga
     2221 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact
     2281 gtacaaatga ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2341 tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     2401 gggcctttct gcgtttatag gtctccgttt tagagctaga aatagcaagt taaaataagg
     2461 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttta ttttttgtca
     2521 ctattgttat gtaaaatgcc acctctgaca gtatggaacg caaacttctg tctagtggat
     2581 ataggtctag agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
     2641 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
     2701 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
     2761 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
     2821 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
     2881 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
     2941 aggatacagt tctcacatca catccgaaca taaacaacaa tgggtaagga aaagactcac
     3001 gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     3061 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     3121 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     3181 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     3241 actcctgatg atgcatggtt actcaccact gcgatccccg gcaaaacagc attccaggta
     3301 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3361 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctg
     3421 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3481 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca
     3541 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3601 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3661 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3721 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3781 tttttctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat
     3841 agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt
     3901 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     3961 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc
     4021 cagtgtcgaa aacgagctct cgagaaccct taatcaaaaa ggatcttcac ctagatcctt
     4081 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4141 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4261 cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata
     4321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4441 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4981 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5101 gttcgtcgac agtcgaacaa gaagcaggca aagtttagag cactgcccct ccgcactcaa
     5161 aaaagaaaaa actaggagga aaataaaatt ctcaaccaca caaacacata aacacataca
     5221 aatacaaata caagcttatt tacttgacat cgcgcgatct tccactattc agcgccgtcc
     5281 gccctctctc gtgttttttg tttacgcgac aactatgcga aatccggagc aacgggcaac
     5341 cgtttgggga aagaccacac ccacgcgcga tcgccatggc aacgaggtcg cacacgcccc
     5401 acacccagac ctccctgcga gcgggcatgg gtacaatgtc cccgttgcca cagacaccac
     5461 ttcgtagcac agcgcagagc gtagcgtgtt gttgctgctg acaaaagaaa atttttctta
     5521 gcaaagcaaa ggaggggaag cacgggcaga tagcaccgta ccataccctt ggaaactcga
     5581 aatgaacgaa gcaggaaatg agagaatgag agttttgtag gtatatatag cggtagtgtt
     5641 tgcgcgttac catcatcttc tggatctatc tattgttctt ttcctcatca ctttcccctt
     5701 tttcgctctt cttcttgtct tttatttctt tctttttttt aattgttccc tcgattggct
     5761 atctaccaaa gaatccaaac ttaatacacg tatttatttg tccaattacc atggataaga
     5821 aatactcaat aggcttagat atcggcacaa atagcgtcgg atgggcggtg atcactgatg
     5881 aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca
     5941 aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgac
     6001 tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg
     6061 agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt
     6121 cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag
     6181 atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag
     6241 attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt
     6301 ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac
     6361 tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa
     6421 gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa
     6481 atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt
     6541 tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat
     6601 tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg caaattggag
     6661 atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag
     6721 atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac
     6781 gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc
     6841 cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg
     6901 atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg
     6961 atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga
     7021 cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga
     7081 gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct
     7141 tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat
     7201 ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata
     7261 aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa
     7321 atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat
     7381 tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac
     7441 agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat
     7501 taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg
     7561 aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata
     7621 aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga
     7681 ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg
     7741 atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc
     7801 gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta gattttttga
     7861 aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat
     7921 ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata
     7981 ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg
     8041 ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg
     8101 cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa
     8161 tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata
     8221 ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg
     8281 tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatcac attgttccac
     8341 aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc
     8401 gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga
     8461 gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg
     8521 aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa
     8581 ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg
     8641 atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt
     8701 ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg
     8761 cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac
     8821 ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta
     8881 agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga
     8941 acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg
     9001 aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc
     9061 gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg
     9121 gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa
     9181 aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc
     9241 tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac
     9301 tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag
     9361 ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt
     9421 ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa
     9481 atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa
     9541 agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc
     9601 attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag
     9661 atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg
     9721 aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt
     9781 ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag
     9841 atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc
     9901 agctaggagg tgacggtgga gggccaaaaa agaaaagaaa agttgaagat gcttctgctc
     9961 atcatcacca tcaccatcac cattaaatcc gctctaaccg aaaaggaagg agttagacaa
    10021 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
    10081 atatttcaaa tttttctttt tttt
//
