LOCUS       sequence-329407-       11551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Combined sgRNA/Cas9 pLSB vector with natMX6 (cloNAT) marker. Golden 
            Gate-ready for cloning custom sgRNA sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11551)
  AUTHORS   Torres-Garcia S, Di Pompeo L, Eivers L, Gaborieau B, White SA, 
            Pidoux AL, Kanigowska P, Yaseen I, Cai Y, Allshire RC
  TITLE     SpEDIT: A fast and efficient CRISPR/Cas9 method for fission yeast.
  JOURNAL   Wellcome Open Res. 2020 Nov 24;5:274. doi: 
            10.12688/wellcomeopenres.16405.1. eCollection 2020.
  PUBMED    33313420
REFERENCE   2  (bases 1 to 11551)
  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: "Wellcome 
            Open Res."; date: "2020-11-24"; volume: "5:274. doi"; pages: " 
            10.12688/wellcomeopenres.16405.1. eCollection 2020"
COMMENT     Sequence Label: pLSB-NAT
FEATURES             Location/Qualifiers
     source          1..11551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    6..27
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(144..161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(315..903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(395..414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1074..1865,1866..1934))
                     /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     1697..1716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1935..2039)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2072..2813
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             2834..6937
                     /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             2834..6937
                     /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             6950..6970
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6988..7235
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7004..7022)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     7282..7300
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7338..7360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7460..7479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7850..8631
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     gene            8705..9824
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        8705..9048
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             9049..9621
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      9627..9824
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     9999..10021
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10014..10030
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10034..10050
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             10468..10479
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             10486..11199
                     /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      11211..11282
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11298..11325
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        11339..11414
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      11424..11443
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(11444..11460)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11464..11480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11477..11499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11488..11504
                     /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(11512..11518,11519..11536,11537..11542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
       61 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
      901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
      961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1201 gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2041 acatttcccc gaaaagtgcc acctgacgca tgccctacaa caactaagaa aatggctatc
     2101 atgcggaagc tggtgagaag aacagcatcg ggacaaggga aggaagaaca aagacaaaga
     2161 aaacaaaaga aagcaattga aaacaaaaca aaacaatttt cattccttct cttatcattc
     2221 cttttctttt cttttctctc attcaacgca ctccatcgta tccgtattcc tcttattttt
     2281 tctctttctc tatatccatt tctttctctc taggtgtgtc ctctctctct cttcaatttc
     2341 tctactccgc attccaacgc atccttcccc caacctccca tttcctcctt acggcccgat
     2401 agcgatcgtc tttccctcgc tatcactcgc taccggcccc tcctctgcac cgtaacctcc
     2461 tacgtattta ccatatcata aagttttttc cgacgcttat cgctgacccc ctgtcgccct
     2521 cctattggct tccggattat cttcttgtcc ataaggtgat ccatgcttcc tgaagattcc
     2581 cgaaatgtgt ccactttggc ggggaatcat tccatccact tctttctctc tcgctttcct
     2641 cattcggcgc tccccttccg cgtctcattg gtcttccgct ccgtttttgc tttgccgatg
     2701 ttacttgggg agaggtgcga taatcctttc gcaaaaactc ggtttgacgc ctcccatggt
     2761 aaatatagtg ggtggtggac aggtgccttc gcttttcttt aagcaagaga attgctgcag
     2821 gtcgactcta gagatggata agaaatactc tattggcctt gatatcggaa ctaacagtgt
     2881 aggttgggca gtcataacag atgaatacaa agttccatct aagaaattta aagtccttgg
     2941 aaacactgac cgacatagta taaaaaagaa tttaatcggc gcattgcttt tcgactctgg
     3001 tgaaacggca gaagccaccc gattgaaacg aacagcaaga cgtcgatata cgcgaagaaa
     3061 gaaccgtatt tgctatcttc aggagatttt ttctaatgaa atggctaagg tcgacgactc
     3121 ctttttccac agattggagg aaagtttcct tgtcgaagag gacaaaaaac acgagcgtca
     3181 ccctatcttc ggaaacattg tagacgaagt ggcatatcat gaaaagtatc ctacgatata
     3241 ccatttgcgt aaaaagttgg tcgactcaac ggacaaggca gatttgcgtt tgatatactt
     3301 ggctcttgcc cacatgatta aatttcgtgg ccatttctta attgaaggtg accttaaccc
     3361 agataacagt gatgtggaca agttgttcat tcaattggtt cagacctaca atcaattatt
     3421 tgaggaaaac ccaatcaatg catctggcgt agatgcaaag gctattcttt ccgcacgcct
     3481 ttcaaaatca cgccgattag agaatcttat cgctcagtta cctggcgaga agaaaaatgg
     3541 cttgttcggt aatttgatcg ctttgagttt gggccttaca ccaaatttta agtccaactt
     3601 cgatcttgca gaagatgcca aattacaatt aagtaaggat acgtacgacg atgatttgga
     3661 caatttactt gcacaaatcg gtgaccaata tgccgatctt ttcttggcag ctaaaaattt
     3721 atcagacgca attttattgt ccgatatcct tcgtgtgaac accgaaatta ccaaggcccc
     3781 acttagtgct tccatgataa aacgctatga tgagcatcat caggacttaa cattacttaa
     3841 agccttggtg cgccaacaat tacccgaaaa gtacaaagaa atattcttcg accagtccaa
     3901 gaacggttat gccggctaca tcgatggtgg tgcatcacag gaggagttct acaaattcat
     3961 taaaccaatc ttagaaaaaa tggatggtac cgaagagtta ttggtaaagt taaatcgcga
     4021 agacttgctt cgcaaacaac gaacattcga caacggctcc atccctcatc aaattcacct
     4081 tggcgaatta catgccatct tacgcagaca agaagacttt tatccattct tgaaggataa
     4141 ccgcgagaaa attgagaaga tccttacctt tagaattcca tactacgtag gccctttggc
     4201 tagaggcaac tccagatttg catggatgac cagaaaatct gaggaaacca tcacgccctg
     4261 gaacttcgaa gaagtcgtag ataagggcgc aagtgcacag tcctttattg aaagaatgac
     4321 caattttgat aagaacttac ctaacgaaaa ggtcttacca aagcatagtt tactttatga
     4381 atacttcacg gtgtacaacg aattaacaaa agtcaaatac gttactgagg gaatgagaaa
     4441 gcctgcattc ttgtccggag aacaaaagaa ggccatagtt gaccttcttt tcaaaacgaa
     4501 tcgaaaggtc acggtaaaac aacttaagga ggattacttt aagaagatag aatgctttga
     4561 tagtgttgag atatcaggtg ttgaagaccg attcaatgca tcattgggaa cgtaccacga
     4621 ccttttgaag atcataaaag ataaagattt tttagacaat gaggagaacg aagacatttt
     4681 agaagacatc gttttgacac ttacgttatt tgaggacaga gagatgatcg aggagcgtct
     4741 taaaacgtat gcacatttat tcgacgacaa agtgatgaaa caacttaagc gcagaagata
     4801 taccggttgg ggaagacttt ctcgaaagtt gatcaacggc atccgcgata agcaaagtgg
     4861 aaaaaccatt ttggatttct taaagtctga tggctttgct aatcgcaact tcatgcaatt
     4921 gattcatgac gattcattaa ctttcaagga agacatacag aaggcccaag tttcaggtca
     4981 gggagatagt ttgcacgagc acatcgctaa tttagctggt tcccctgcta taaaaaaggg
     5041 tatacttcaa accgtaaaag tcgtagatga gttggtaaaa gtcatgggca gacataaacc
     5101 agaaaatata gtaattgaga tggcccgaga aaatcaaacg acgcaaaagg gccaaaaaaa
     5161 ctctagagaa agaatgaagc gtatcgagga gggcataaag gagttgggtt ctcagatatt
     5221 gaaggagcac cccgttgaga acactcagct tcagaatgag aagctttatt tatattattt
     5281 acaaaacggt cgtgatatgt atgtggatca ggagttggac ataaacagat tgtcagatta
     5341 cgatgtggat catatcgtcc cccaatcatt cttaaaggac gattcaattg acaacaaagt
     5401 tcttacgcgt agtgacaaga acagaggcaa atcagacaat gtacctagtg aggaagtagt
     5461 gaaaaagatg aagaactatt ggagacaact tcttaatgct aagttaataa cgcagcgaaa
     5521 gtttgataac ttaactaaag ccgaacgtgg cggactttca gagttggata aagcaggctt
     5581 tatcaaacgt caattggtag agactcgtca gatcactaag catgtggccc aaatacttga
     5641 ttcacgcatg aatactaagt acgatgaaaa cgataaattg attcgtgagg ttaaggtaat
     5701 cacgttgaaa tctaagcttg tttccgactt tcgtaaggat tttcaattct acaaggttcg
     5761 agagattaat aattaccacc atgcacatga cgcatacttg aacgcagtcg taggtacagc
     5821 attgatcaag aagtacccaa aattagagtc agaatttgtc tacggtgact acaaagtata
     5881 tgatgtccga aagatgatag ccaagtctga gcaagagata ggaaaggcca cagctaagta
     5941 ttttttttac tccaacatta tgaatttctt taagacagag attactttag ctaacggtga
     6001 aatacgaaaa agacccctta tcgaaacgaa tggagagaca ggtgagattg tttgggacaa
     6061 aggtcgagat ttcgccactg tgagaaaagt cttgagtatg ccacaggtca atattgttaa
     6121 gaaaacagaa gtacagactg gtggattttc taaagaatcc attttgccca agagaaattc
     6181 cgacaaactt attgcacgca agaaggattg ggatcccaag aaatacggcg gtttcgacag
     6241 tcccactgta gcatactcag tgttggtcgt ggctaaggtg gaaaagggta agtctaagaa
     6301 gcttaaatct gtgaaggaat tgcttggtat cacgatcatg gagcgctcat cttttgagaa
     6361 aaatcctata gactttcttg aagctaaagg ctataaagag gtgaaaaagg accttataat
     6421 taagttacca aagtactctc ttttcgaact tgagaacgga cgcaagagaa tgcttgccag
     6481 tgctggagaa ttacagaagg gtaatgagtt agcattgcca tcaaagtacg taaacttctt
     6541 atatttagcc tcccactatg agaaattaaa gggttcaccc gaggacaatg aacaaaagca
     6601 attattcgta gaacaacata aacactattt ggacgaaatt atagagcaaa tctctgaatt
     6661 ttccaaaaga gtgattttag ctgacgcaaa tcttgataag gttttatctg cctataacaa
     6721 gcatcgcgac aaacccatca gagaacaagc cgaaaatatc attcacttat tcacacttac
     6781 gaatcttgga gcccctgccg ctttcaagta ttttgatact accatagacc gtaaacgcta
     6841 tacgtctact aaggaagtct tagacgccac tcttattcac caaagtataa ccggcttgta
     6901 tgagacaaga attgatttgt ctcaattggg cggcgacagc agggctgacc ccaagaagaa
     6961 gaggaaggtg tgattttgga cctcgagtca tgtaattagt tatgtcacgc ttacattcac
     7021 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     7081 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     7141 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     7201 agaaggtttt gggacgctcg aaggctttaa tttgcgatcc ccgtctaaga aaccattatt
     7261 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     7321 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     7381 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     7441 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataccatc
     7501 gatggccaac cttccaattc attaaatcga gtcctccatc agatgattta caaggttttg
     7561 catagaatcc ctaagtaaga gaagttgtta gtgatttctt caagtttact aaaatttaca
     7621 tacaaatgga tgatctcttc tccattgttt cctttcttct tgcaaacgcg ttttacaaag
     7681 agatgacatg ctattttctg attatttttt tctatagttt tctattttgt acgtccttgg
     7741 gagtccgaaa tgtaaaatcg gggtatggtt agtagcggca gcgttcatta tggtatataa
     7801 aagataaaaa gtaggggcaa gcgagacaga atgggataca agggcatcgt ctataattat
     7861 agctaaaaat tgtattttaa tttgtatttt ttgtaatttt atttctcatt gttttactta
     7921 aaatgaatgc gaattagaga aaacttcaac gaaatgtcaa aataagctca gcaaaatata
     7981 caattttagg gaaagcgatc agcaacttat tctccgttgt agattctttt tcttcataaa
     8041 ttccaacaat gtaactgcca attgattttt ctctagatgc taataaattg aaactaatga
     8101 atataataac acaggttttg ataaattgta aactcttact caaaaaaaga atgttaaaac
     8161 taatttaacc tttttataaa ggctcatggc gttaaagagg atagtaaaaa ttctacaatt
     8221 ttttaattta atgtcacaat tgtcttgcga atataattct attaattttc gctaactgct
     8281 gttttagtac tattactttc aatatgcctt atatgcaact ttaattcatg aagaatgcaa
     8341 tatttttgca atctttcttg tactgagcgc tttttatcat ttagtactca ttatttaatt
     8401 tttcagtaaa gggcaataaa taaatttgta gaagatgcaa tgtaatctcc tctatccttt
     8461 tgctcatatg tttatgagta tacctagtct agaaaggctt gtatttaaaa tattattcaa
     8521 taaaaattca caatttttac gacatgtgct accattcact taacttcctg attataaaat
     8581 tggttcgttt atactaatta cttaagtacc tttaactaaa caaaatgcct atatatatat
     8641 taatttacaa tgagtgtcag ataagtcact atgtccgagt ggttaaggag ttagactcgc
     8701 catcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     8761 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     8821 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     8881 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     8941 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     9001 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaccact
     9061 cttgacgaca cggcttaccg gtaccgcacc agtgtcccgg gggacgccga ggccatcgag
     9121 gcactggatg ggtccttcac caccgacacc gtcttccgcg tcaccgccac cggggacggc
     9181 ttcaccctgc gggaggtgcc ggtggacccg cccctgacca aggtgttccc cgacgacgaa
     9241 tcggacgacg aatcggacga cggggaggac ggcgacccgg actcccggac gttcgtcgcg
     9301 tacggggacg acggcgacct ggcgggcttc gtggtcgtct cgtactccgg ctggaaccgc
     9361 cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg ggtcgggcgc
     9421 gcgttgatgg ggctcgcgac ggagttcgcc cgcgagcggg gcgccgggca cctctggctg
     9481 gaggtcacca acgtcaacgc accggcgatc cacgcgtacc ggcggatggg gttcaccctc
     9541 tgcggcctgg acaccgccct gtacgacggc accgcctcgg acggcgagca ggcgctctac
     9601 atgagcatgc cctgccccta atcagtactg acaataaaaa gattcttgtt ttcaagaact
     9661 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     9721 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     9781 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgggtgtg aaataccgca
     9841 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     9901 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     9961 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
    10021 gacggccagt gttgtaaaac gacggccagt accatttgag cattggaaac ttcctatcaa
    10081 aaattacata gaaatttttg attttggcta tataaattac atttcatgat aaatgtacag
    10141 ctaaagtcac tatgtccgag tggttaagga gttagacttc aatcctgtat tctagtcatc
    10201 taatgggctt tgcccgcgca ggttcaaatc ctgctggtga cggatggccg gcatggtccc
    10261 agcctcctcg ctggcgccgg ctgggcaaca ccttcgggtg gcgaatggga ctagagaccg
    10321 aaagtgaaac gtgatttcat gcgtcatttt gaacattttg taaatcttat ttaataatgt
    10381 gtgcggcaat tcacatttaa tttatgaatg ttttcttaac atcgcggcaa ctcaagaaac
    10441 ggcaggttcg gatcttagct actagagaaa gaggagaaat actagatgcg taaaggcgaa
    10501 gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat
    10561 aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag
    10621 ttcatctgta ctactggtaa actgccggtt ccttggccga ctctggtaac gacgctgact
    10681 tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag
    10741 tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg
    10801 tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg
    10861 aaaggcattg actttaaaga ggacggcaat atcctgggcc ataagctgga atacaatttt
    10921 aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt
    10981 aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac
    11041 actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc
    11101 gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc
    11161 gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat gaccaggcat caaataaaac
    11221 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
    11281 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat aggtctcagt
    11341 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
    11401 caccgagtcg gtgctttttc attttttttt gttttttatg tctgtcatag ctgtttcctg
    11461 tgtgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    11521 agccggaagc ataaagtgta aagcctgggg t
//
