LOCUS       sequence-467135-       10323 bp    DNA     circular SYN 15-APR-2025
DEFINITION  SpCAS9-Tyr-[gRNA: BsaI GFP dropout] with pan(OPT)ARS replication 
            origin and Hyg selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10323)
  TITLE     Sequencing of a dairy isolate unlocks Kluyveromyces marxianus as a 
            host for lactose valorization
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10323)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pMTKM05
FEATURES             Location/Qualifiers
     source          1..10323
                     /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..4211
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2636..2978
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2980..4008
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      4014..4211
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4362..5153,5154..5222))
                     /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     4985..5004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5223..5327)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6030..10133
                     /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             6030..10133
                     /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             10143..10163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10179..10202
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      join(10206..10323,1..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 tgggaaagaa accagaattg
     3001 acagctacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtgtc cgacctgatg
     3061 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     3121 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     3181 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt tagcgagagc
     3241 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     3301 gaactgcccg ctgttctgca accggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     3361 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     3421 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     3481 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     3541 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     3601 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     3661 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     3721 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     3781 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     3841 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     3901 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     3961 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataatc agtactgaca
     4021 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     4081 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     4141 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     4201 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa acgagctctc
     4261 gagaaccctt aatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4501 cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttcgtcgaca gtcgaacaag
     5341 aagcaggcaa agtttagagc actgcccctc cgcactcaaa aaagaaaaaa ctaggaggaa
     5401 aataaaattc tcaaccacac aaacacataa acacatacaa atacaaatac aagcttattt
     5461 acttgacatc gcgcgatctt ccactattca gcgccgtccg ccctctctcg tgttttttgt
     5521 ttacgcgaca actatgcgaa atccggagca acgggcaacc gtttggggaa agaccacacc
     5581 cacgcgcgat cgccatggca acgaggtcgc acacgcccca cacccagacc tccctgcgag
     5641 cgggcatggg tacaatgtcc ccgttgccac agacaccact tcgtagcaca gcgcagagcg
     5701 tagcgtgttg ttgctgctga caaaagaaaa tttttcttag caaagcaaag gaggggaagc
     5761 acgggcagat agcaccgtac catacccttg gaaactcgaa atgaacgaag caggaaatga
     5821 gagaatgaga gttttgtagg tatatatagc ggtagtgttt gcgcgttacc atcatcttct
     5881 ggatctatct attgttcttt tcctcatcac tttccccttt ttcgctcttc ttcttgtctt
     5941 ttatttcttt ctttttttta attgttccct cgattggcta tctaccaaag aatccaaact
     6001 taatacacgt atttatttgt ccaattacca tggataagaa atactcaata ggcttagata
     6061 tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa
     6121 agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc
     6181 ttttatttga cagtggagag acagcggaag cgactcgact caaacggaca gctcgtagaa
     6241 ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg
     6301 cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca
     6361 agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga
     6421 aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt
     6481 tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg
     6541 agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa
     6601 cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga
     6661 ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg
     6721 gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta
     6781 attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt
     6841 acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt
     6901 tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg
     6961 aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag
     7021 acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct
     7081 tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag
     7141 aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg
     7201 tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc
     7261 cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc
     7321 catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt
     7381 atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag
     7441 aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat
     7501 ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac
     7561 atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta
     7621 ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt
     7681 tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa
     7741 aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat
     7801 taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag
     7861 aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga
     7921 tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc
     7981 ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt aatggtatta
     8041 gggataagca atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc
     8101 gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac attcaaaaag
     8161 cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc
     8221 ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa
     8281 tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc
     8341 aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat
     8401 taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc
     8461 tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta
     8521 atcgtttaag tgattatgat gtcgatcaca ttgttccaca aagtttcctt aaagacgatt
     8581 caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc
     8641 caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt
     8701 taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt ttgagtgaac
     8761 ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg
     8821 tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc
     8881 gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc
     8941 aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg
     9001 ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg
     9061 gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca
     9121 aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta
     9181 cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag
     9241 aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc
     9301 aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag gagtcaattt
     9361 taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat
     9421 atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa
     9481 aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa
     9541 gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta
     9601 aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta
     9661 aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca
     9721 aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag
     9781 ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg
     9841 agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc
     9901 ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc
     9961 atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa
    10021 ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat
    10081 ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt gacggtggag
    10141 ggccaaaaaa gaaaagaaaa gttgaagatg cttctgctca tcatcaccat caccatcacc
    10201 attaaatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
    10261 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
    10321 ttt
//
