LOCUS       sequence-329224-       11788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Combined sgRNA/Cas9 pLSB vector with kanMX6 (G418) 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 11788)
  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 11788)
  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-KAN
FEATURES             Location/Qualifiers
     source          1..11788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..50)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(204..792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(284..303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(963..1754,1755..1823))
                     /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     1586..1605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1824..1928)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1961..2702
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             2723..6826
                     /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             2723..6826
                     /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             6839..6859
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6877..7124
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6893..6911)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     7171..7189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7227..7249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7349..7368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7739..8520
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     gene            8594..9950
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        8594..8937
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             8938..9747
                     /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(9005..9024)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      9753..9950
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     10125..10147
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10140..10156
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10160..10176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             10594..10605
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             10612..11325
                     /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      11337..11408
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11424..11451
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        11465..11540
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      11550..11569
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(11570..11586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11590..11606)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11603..11625)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11614..11630
                     /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(11638..11644,11645..11662,11663..11668))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11683..11704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
       61 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
      961 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1021 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1081 cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata
     1141 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1201 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1261 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1321 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1381 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1441 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1501 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1561 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1621 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1681 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1741 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1801 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1861 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1921 gttccgcgca catttccccg aaaagtgcca cctgacgcat gccctacaac aactaagaaa
     1981 atggctatca tgcggaagct ggtgagaaga acagcatcgg gacaagggaa ggaagaacaa
     2041 agacaaagaa aacaaaagaa agcaattgaa aacaaaacaa aacaattttc attccttctc
     2101 ttatcattcc ttttcttttc ttttctctca ttcaacgcac tccatcgtat ccgtattcct
     2161 cttatttttt ctctttctct atatccattt ctttctctct aggtgtgtcc tctctctctc
     2221 ttcaatttct ctactccgca ttccaacgca tccttccccc aacctcccat ttcctcctta
     2281 cggcccgata gcgatcgtct ttccctcgct atcactcgct accggcccct cctctgcacc
     2341 gtaacctcct acgtatttac catatcataa agttttttcc gacgcttatc gctgaccccc
     2401 tgtcgccctc ctattggctt ccggattatc ttcttgtcca taaggtgatc catgcttcct
     2461 gaagattccc gaaatgtgtc cactttggcg gggaatcatt ccatccactt ctttctctct
     2521 cgctttcctc attcggcgct ccccttccgc gtctcattgg tcttccgctc cgtttttgct
     2581 ttgccgatgt tacttgggga gaggtgcgat aatcctttcg caaaaactcg gtttgacgcc
     2641 tcccatggta aatatagtgg gtggtggaca ggtgccttcg cttttcttta agcaagagaa
     2701 ttgctgcagg tcgactctag agatggataa gaaatactct attggccttg atatcggaac
     2761 taacagtgta ggttgggcag tcataacaga tgaatacaaa gttccatcta agaaatttaa
     2821 agtccttgga aacactgacc gacatagtat aaaaaagaat ttaatcggcg cattgctttt
     2881 cgactctggt gaaacggcag aagccacccg attgaaacga acagcaagac gtcgatatac
     2941 gcgaagaaag aaccgtattt gctatcttca ggagattttt tctaatgaaa tggctaaggt
     3001 cgacgactcc tttttccaca gattggagga aagtttcctt gtcgaagagg acaaaaaaca
     3061 cgagcgtcac cctatcttcg gaaacattgt agacgaagtg gcatatcatg aaaagtatcc
     3121 tacgatatac catttgcgta aaaagttggt cgactcaacg gacaaggcag atttgcgttt
     3181 gatatacttg gctcttgccc acatgattaa atttcgtggc catttcttaa ttgaaggtga
     3241 ccttaaccca gataacagtg atgtggacaa gttgttcatt caattggttc agacctacaa
     3301 tcaattattt gaggaaaacc caatcaatgc atctggcgta gatgcaaagg ctattctttc
     3361 cgcacgcctt tcaaaatcac gccgattaga gaatcttatc gctcagttac ctggcgagaa
     3421 gaaaaatggc ttgttcggta atttgatcgc tttgagtttg ggccttacac caaattttaa
     3481 gtccaacttc gatcttgcag aagatgccaa attacaatta agtaaggata cgtacgacga
     3541 tgatttggac aatttacttg cacaaatcgg tgaccaatat gccgatcttt tcttggcagc
     3601 taaaaattta tcagacgcaa ttttattgtc cgatatcctt cgtgtgaaca ccgaaattac
     3661 caaggcccca cttagtgctt ccatgataaa acgctatgat gagcatcatc aggacttaac
     3721 attacttaaa gccttggtgc gccaacaatt acccgaaaag tacaaagaaa tattcttcga
     3781 ccagtccaag aacggttatg ccggctacat cgatggtggt gcatcacagg aggagttcta
     3841 caaattcatt aaaccaatct tagaaaaaat ggatggtacc gaagagttat tggtaaagtt
     3901 aaatcgcgaa gacttgcttc gcaaacaacg aacattcgac aacggctcca tccctcatca
     3961 aattcacctt ggcgaattac atgccatctt acgcagacaa gaagactttt atccattctt
     4021 gaaggataac cgcgagaaaa ttgagaagat ccttaccttt agaattccat actacgtagg
     4081 ccctttggct agaggcaact ccagatttgc atggatgacc agaaaatctg aggaaaccat
     4141 cacgccctgg aacttcgaag aagtcgtaga taagggcgca agtgcacagt cctttattga
     4201 aagaatgacc aattttgata agaacttacc taacgaaaag gtcttaccaa agcatagttt
     4261 actttatgaa tacttcacgg tgtacaacga attaacaaaa gtcaaatacg ttactgaggg
     4321 aatgagaaag cctgcattct tgtccggaga acaaaagaag gccatagttg accttctttt
     4381 caaaacgaat cgaaaggtca cggtaaaaca acttaaggag gattacttta agaagataga
     4441 atgctttgat agtgttgaga tatcaggtgt tgaagaccga ttcaatgcat cattgggaac
     4501 gtaccacgac cttttgaaga tcataaaaga taaagatttt ttagacaatg aggagaacga
     4561 agacatttta gaagacatcg ttttgacact tacgttattt gaggacagag agatgatcga
     4621 ggagcgtctt aaaacgtatg cacatttatt cgacgacaaa gtgatgaaac aacttaagcg
     4681 cagaagatat accggttggg gaagactttc tcgaaagttg atcaacggca tccgcgataa
     4741 gcaaagtgga aaaaccattt tggatttctt aaagtctgat ggctttgcta atcgcaactt
     4801 catgcaattg attcatgacg attcattaac tttcaaggaa gacatacaga aggcccaagt
     4861 ttcaggtcag ggagatagtt tgcacgagca catcgctaat ttagctggtt cccctgctat
     4921 aaaaaagggt atacttcaaa ccgtaaaagt cgtagatgag ttggtaaaag tcatgggcag
     4981 acataaacca gaaaatatag taattgagat ggcccgagaa aatcaaacga cgcaaaaggg
     5041 ccaaaaaaac tctagagaaa gaatgaagcg tatcgaggag ggcataaagg agttgggttc
     5101 tcagatattg aaggagcacc ccgttgagaa cactcagctt cagaatgaga agctttattt
     5161 atattattta caaaacggtc gtgatatgta tgtggatcag gagttggaca taaacagatt
     5221 gtcagattac gatgtggatc atatcgtccc ccaatcattc ttaaaggacg attcaattga
     5281 caacaaagtt cttacgcgta gtgacaagaa cagaggcaaa tcagacaatg tacctagtga
     5341 ggaagtagtg aaaaagatga agaactattg gagacaactt cttaatgcta agttaataac
     5401 gcagcgaaag tttgataact taactaaagc cgaacgtggc ggactttcag agttggataa
     5461 agcaggcttt atcaaacgtc aattggtaga gactcgtcag atcactaagc atgtggccca
     5521 aatacttgat tcacgcatga atactaagta cgatgaaaac gataaattga ttcgtgaggt
     5581 taaggtaatc acgttgaaat ctaagcttgt ttccgacttt cgtaaggatt ttcaattcta
     5641 caaggttcga gagattaata attaccacca tgcacatgac gcatacttga acgcagtcgt
     5701 aggtacagca ttgatcaaga agtacccaaa attagagtca gaatttgtct acggtgacta
     5761 caaagtatat gatgtccgaa agatgatagc caagtctgag caagagatag gaaaggccac
     5821 agctaagtat tttttttact ccaacattat gaatttcttt aagacagaga ttactttagc
     5881 taacggtgaa atacgaaaaa gaccccttat cgaaacgaat ggagagacag gtgagattgt
     5941 ttgggacaaa ggtcgagatt tcgccactgt gagaaaagtc ttgagtatgc cacaggtcaa
     6001 tattgttaag aaaacagaag tacagactgg tggattttct aaagaatcca ttttgcccaa
     6061 gagaaattcc gacaaactta ttgcacgcaa gaaggattgg gatcccaaga aatacggcgg
     6121 tttcgacagt cccactgtag catactcagt gttggtcgtg gctaaggtgg aaaagggtaa
     6181 gtctaagaag cttaaatctg tgaaggaatt gcttggtatc acgatcatgg agcgctcatc
     6241 ttttgagaaa aatcctatag actttcttga agctaaaggc tataaagagg tgaaaaagga
     6301 ccttataatt aagttaccaa agtactctct tttcgaactt gagaacggac gcaagagaat
     6361 gcttgccagt gctggagaat tacagaaggg taatgagtta gcattgccat caaagtacgt
     6421 aaacttctta tatttagcct cccactatga gaaattaaag ggttcacccg aggacaatga
     6481 acaaaagcaa ttattcgtag aacaacataa acactatttg gacgaaatta tagagcaaat
     6541 ctctgaattt tccaaaagag tgattttagc tgacgcaaat cttgataagg ttttatctgc
     6601 ctataacaag catcgcgaca aacccatcag agaacaagcc gaaaatatca ttcacttatt
     6661 cacacttacg aatcttggag cccctgccgc tttcaagtat tttgatacta ccatagaccg
     6721 taaacgctat acgtctacta aggaagtctt agacgccact cttattcacc aaagtataac
     6781 cggcttgtat gagacaagaa ttgatttgtc tcaattgggc ggcgacagca gggctgaccc
     6841 caagaagaag aggaaggtgt gattttggac ctcgagtcat gtaattagtt atgtcacgct
     6901 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     6961 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     7021 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     7081 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcgatccc cgtctaagaa
     7141 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7201 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7261 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     7321 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7381 cataccatcg atggccaacc ttccaattca ttaaatcgag tcctccatca gatgatttac
     7441 aaggttttgc atagaatccc taagtaagag aagttgttag tgatttcttc aagtttacta
     7501 aaatttacat acaaatggat gatctcttct ccattgtttc ctttcttctt gcaaacgcgt
     7561 tttacaaaga gatgacatgc tattttctga ttattttttt ctatagtttt ctattttgta
     7621 cgtccttggg agtccgaaat gtaaaatcgg ggtatggtta gtagcggcag cgttcattat
     7681 ggtatataaa agataaaaag taggggcaag cgagacagaa tgggatacaa gggcatcgtc
     7741 tataattata gctaaaaatt gtattttaat ttgtattttt tgtaatttta tttctcattg
     7801 ttttacttaa aatgaatgcg aattagagaa aacttcaacg aaatgtcaaa ataagctcag
     7861 caaaatatac aattttaggg aaagcgatca gcaacttatt ctccgttgta gattcttttt
     7921 cttcataaat tccaacaatg taactgccaa ttgatttttc tctagatgct aataaattga
     7981 aactaatgaa tataataaca caggttttga taaattgtaa actcttactc aaaaaaagaa
     8041 tgttaaaact aatttaacct ttttataaag gctcatggcg ttaaagagga tagtaaaaat
     8101 tctacaattt tttaatttaa tgtcacaatt gtcttgcgaa tataattcta ttaattttcg
     8161 ctaactgctg ttttagtact attactttca atatgcctta tatgcaactt taattcatga
     8221 agaatgcaat atttttgcaa tctttcttgt actgagcgct ttttatcatt tagtactcat
     8281 tatttaattt ttcagtaaag ggcaataaat aaatttgtag aagatgcaat gtaatctcct
     8341 ctatcctttt gctcatatgt ttatgagtat acctagtcta gaaaggcttg tatttaaaat
     8401 attattcaat aaaaattcac aatttttacg acatgtgcta ccattcactt aacttcctga
     8461 ttataaaatt ggttcgttta tactaattac ttaagtacct ttaactaaac aaaatgccta
     8521 tatatatatt aatttacaat gagtgtcaga taagtcacta tgtccgagtg gttaaggagt
     8581 tagactcgcc atcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
     8641 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
     8701 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
     8761 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
     8821 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
     8881 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
     8941 ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga tgctgattta
     9001 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     9061 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     9121 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     9181 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
     9241 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     9301 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     9361 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     9421 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     9481 aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     9541 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     9601 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     9661 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     9721 catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt cttgttttca
     9781 agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta
     9841 gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc
     9901 agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ggtgtgaaat
     9961 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
    10021 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
    10081 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
    10141 taaaacgacg gccagtgttg taaaacgacg gccagtacca tttgagcatt ggaaacttcc
    10201 tatcaaaaat tacatagaaa tttttgattt tggctatata aattacattt catgataaat
    10261 gtacagctaa agtcactatg tccgagtggt taaggagtta gacttcaatc ctgtattcta
    10321 gtcatctaat gggctttgcc cgcgcaggtt caaatcctgc tggtgacgga tggccggcat
    10381 ggtcccagcc tcctcgctgg cgccggctgg gcaacacctt cgggtggcga atgggactag
    10441 agaccgaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa tcttatttaa
    10501 taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg cggcaactca
    10561 agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta gatgcgtaaa
    10621 ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
    10681 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
    10741 ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct ggtaacgacg
    10801 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
    10861 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
    10921 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
    10981 gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa gctggaatac
    11041 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
    11101 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
    11161 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
    11221 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
    11281 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc aggcatcaaa
    11341 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
    11401 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttataggt
    11461 ctcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
    11521 aagtggcacc gagtcggtgc tttttcattt ttttttgttt tttatgtctg tcatagctgt
    11581 ttcctgtgtg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    11641 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    11701 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    11761 ttaatgaatc ggccaacgcg cggggaga
//
