LOCUS       sequence-351363-       15413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS414 backbone containing LINEAR fragment targeting XYL2 in S. 
            stipitis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15413)
  AUTHORS   Ploessl D, Zhao Y, Cao M, Ghosh S, Lopez C, Sayadi M, Chudalayandi 
            S, Severin A, Huang L, Gustafson M, Shao Z
  TITLE     A repackaged CRISPR platform increases homology-directed repair for 
            yeast engineering.
  JOURNAL   Nat Chem Biol. 2021 Oct 28. pii: 10.1038/s41589-021-00893-5. doi: 
            10.1038/s41589-021-00893-5.
  PUBMED    34711982
REFERENCE   2  (bases 1 to 15413)
  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: "Nat Chem 
            Biol. 2021 Oct 28. pii: 10.1038/s41589-021-00893-5. doi: 
            10.1038/s41589-021-00893-5."
COMMENT     Sequence Label: pCashGem-Xyl2-URA3-LINEAR
FEATURES             Location/Qualifiers
     source          1..15413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1762..2265
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2306..2324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2362..2384)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2484..2503
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2524..2804
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             2805..3479
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(3582..4037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3719..3740)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3931..3950
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4163..4185
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4177..4194
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4178..4194
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4204..4223
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4204..4222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5682..5705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5709..5729
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5730..9830
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5730..9830
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9834..9854
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9861..10190
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     CDS             11127..11843
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_RNA        12318..12393
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(14973..14993)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(14973..14991)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(15012..15028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15012..15028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15025..15047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15036..15052
                     /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(15060..15066,15067..15084,15085..15090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15105..15126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15243..15260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta
     1801 atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt
     1861 tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt
     1921 gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca
     1981 cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta
     2041 atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta
     2101 aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc
     2161 taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc
     2221 atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtct aagaaaccat
     2281 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     2341 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2401 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2461 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag
     2521 atcacgacat tactatatat ataatatagg aagcatttaa tagaacagca tcgtaatata
     2581 tgtgtacttt gcagttatga cgccagatgg cagtagtgga agatattctt tattgaaaaa
     2641 tagcttgtca ccttacgtac aatcttgatc cggagctttt ctttttttgc cgattaagaa
     2701 ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg agggcattgg tgactattga
     2761 gcacgtgagt atacgtgatt aagcacacaa aggcagcttg gagtatgtct gttattaatt
     2821 tcacaggtag ttctggtcca ttggtgaaag tttgcggctt gcagagcaca gaggccgcag
     2881 aatgtgctct agattccgat gctgacttgc tgggtattat atgtgtgccc aatagaaaga
     2941 gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata
     3001 gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt
     3061 tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat gagtcgtggc
     3121 aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact
     3181 gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt
     3241 cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa
     3301 ggcaagagag ccccgaaagc ttacatttta tgttagctgg tggactgacg ccagaaaatg
     3361 ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga
     3421 caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg
     3481 ttattactga gtagtattta tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg
     3541 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     3601 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3661 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3721 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3781 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3841 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3901 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3961 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4021 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4081 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     4141 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     4201 gcgtaatacg actcactata gggcgaattg ggtaccgggc cctgcaggga aggtgcacct
     4261 gatgctgaac cagctgaaga tgttgatccg gctgcaacgg tggaactgct ccccggcgcc
     4321 agcttcgttt tggcgctgct gacgtccttg gcttctttgc tggacatatc tgaaacgaga
     4381 atatggcggt gcttactggt tttgcgattg gttgagttgt tgtttccaga atgtaaatac
     4441 tgagagactg tatgagaagt tagaaaccga gaatagtctt agtgaaaatt aatgggaaca
     4501 ggaagtccct tgtgtacgta acagcttatg taataaatgt agaagcctca cccggagctc
     4561 agttccgggc acgcagatca catgaaagtg actgacaagg tgtccgggca gttccgaggg
     4621 cgtccggtga tgaaatagct gcatgcagtg ccctcctttg gcacgaaccg aatgttgacg
     4681 ggcaatggga acttgttgcc tcggcgcacg tgacaaatgt gattatgtcg aaaactccga
     4741 gtccgagagc agacagacgg agtgcggtgg gaaggaacaa tcagatcagg gagaccagcg
     4801 agaatcgtac gctcctctag aactgtgtta ttgagtcatt ggagtggctt caatggatat
     4861 gtgccattgt ggcggttatg aacgttcaga atgcgattga ggggtagagt gaatcatttt
     4921 tgacggtgcc agccagagag gaacgcatgg tgcaagtgaa aaatttcagc tactgtttga
     4981 gagcaattag aatactgtta tgagccaata aactagtcaa accaaatagc caatcaacca
     5041 gccaacaagc cggtggtagc cagccttggg acaggcattg gcacacaaat aacctggtgt
     5101 actgtaacta catcaccagc caccgtatca ctgctccatt atcagtgcca tctcatgagc
     5161 attggagctg ttgatgcaag ctgtcgctaa tatgccgcaa caaattggac tcatttttag
     5221 ggcaattcta tccagtacca ataaagcacg aatcgcttta tgaatcatag cctggccgta
     5281 gcatttcagc aatttcgcag gttatggttt aacagcgacg tacaaaactt ttcacagtca
     5341 tatacggtat acccaaacat ggattcgtgg acttcggctc ctccgttgaa ctcatattcg
     5401 taatccccat tcagattgcc ctctcatgat gcccaccagt tgcaatctgg tgatcgcatt
     5461 atgcacactc ttcgggtatc gggactgagt ggtccagttt cgcacaaaat tcgcacacgg
     5521 tgaacaagat ggcccacact tttttcactc gacatataaa gggaacgaga tttcctcctt
     5581 gatttctcct ggcattgcgt actgtgtatt ttttgcatct agtcaattat ctgatttcca
     5641 gctaattact tgcttcttta tcgattcccg cactaacaat ggactacaag gatgacgatg
     5701 acaagcctcc aaagaaaaag agaaaggttg acaagaagta ctctatcggt ttggacatcg
     5761 gcactaactc tgttggttgg gctgttatca ctgacgagta caaggtccca tccaagaagt
     5821 tcaaggtctt gggtaacacc gacagacact ccatcaagaa gaacttgatc ggcgccttgt
     5881 tgttcgactc tggtgaaact gctgaagcca ccagattgaa gagaaccgcc agaagaagat
     5941 acaccagaag aaagaacaga atctgctact tgcaagagat cttctccaac gaaatggcca
     6001 aggtcgacga ctcgttcttc cacagattgg aagaatcctt cttggtcgaa gaggacaaga
     6061 agcacgaaag acacccaatc ttcggtaaca tcgttgacga agtcgcctac cacgaaaagt
     6121 acccaaccat ctaccacttg agaaagaagt tggtcgactc taccgacaag gccgacttga
     6181 gattgatcta cttggctttg gcccacatga tcaagttcag aggtcacttc ttgatcgagg
     6241 gcgacttgaa cccagataac tctgacgttg acaagttgtt catccagttg gtccagacct
     6301 acaaccagtt gttcgaagaa aacccaatca acgcttctgg tgtcgacgct aaggctatct
     6361 tgtctgccag attgtccaag tccagaagat tggaaaactt gatcgcccag ttgccaggcg
     6421 aaaagaagaa cggtttgttc ggcaacttga tcgctttgtc cttgggcttg acccccaact
     6481 tcaagtctaa cttcgacttg gctgaggacg ccaagttgca gttgtctaag gatacttacg
     6541 acgacgactt ggacaacttg ttggctcaaa tcggtgacca gtacgctgac ttgttcttgg
     6601 ctgctaagaa cttgtctgac gccatcttgt tgtccgacat cttgagagtc aacaccgaaa
     6661 tcaccaaggc cccattgtct gcctctatga tcaagagata cgacgaacac caccaggact
     6721 tgaccttgtt gaaggctttg gttagacagc agttgcccga gaagtacaaa gaaatcttct
     6781 tcgaccagtc caagaacggt tacgctggtt acatcgatgg tggtgcctcc caagaagagt
     6841 tctacaagtt catcaagccc atcttggaaa agatggacgg caccgaagag ttgttggtca
     6901 agttgaacag agaggacttg ttgagaaagc agagaacctt cgacaacggt tctatcccac
     6961 atcagatcca cttgggtgaa ttgcacgcca tcttgagaag acaagaggac ttctacccat
     7021 tcttgaagga caacagagaa aagatcgaga agatcttgac cttcagaatc ccctactacg
     7081 tcggtccatt ggccagaggt aattctagat tcgcctggat gaccagaaag tccgaagaaa
     7141 ctatcacccc ctggaacttc gaagaagttg ttgataaggg cgcctccgcc cagtctttca
     7201 tcgaaagaat gaccaacttc gacaagaact tgcccaacga gaaggtcttg ccaaagcact
     7261 ccttgttgta cgagtacttc accgtctaca acgagttgac caaggtcaag tacgtcaccg
     7321 aaggtatgag aaagccagcc ttcttgtctg gtgaacagaa gaaggctatc gtcgacttgt
     7381 tgttcaagac caacagaaag gtcaccgtca agcagttgaa agaggactac ttcaagaaaa
     7441 tcgagtgctt cgactccgtc gaaatctctg gtgtcgaaga tagattcaac gcctccttgg
     7501 gtacttacca cgacttgttg aagatcatca aggacaagga tttcttggac aacgaagaga
     7561 acgaggacat cttggaggac atcgtcttga ctttgacctt gttcgaggac agagagatga
     7621 tcgaagagag attgaaaacc tacgcccact tgttcgacga caaggtcatg aagcagttga
     7681 agagaagaag atataccggc tggggcagat tgtccagaaa gttgatcaac ggtatcagag
     7741 acaagcagtc cggcaagact atcttggact tcttgaagtc tgacggcttc gccaacagaa
     7801 acttcatgca gttgatccac gacgattcct tgaccttcaa agaggacatc cagaaggctc
     7861 aagtttctgg tcaaggtgac tccttgcacg aacatatcgc taacttggct ggttctccag
     7921 ccatcaagaa gggtatcttg cagactgtta aggtcgtcga cgaattggtg aaggtcatgg
     7981 gtagacacaa gccagagaac atcgtcatcg aaatggctag agaaaaccag accacccaga
     8041 agggtcagaa gaactccaga gaaagaatga agagaatcga agagggcatc aaagagttgg
     8101 gctcccagat cttgaaagaa cacccagtcg aaaacaccca gttgcagaac gagaagttgt
     8161 acttgtacta cttgcagaac ggcagagaca tgtacgtcga ccaagagttg gacatcaaca
     8221 gattgtctga ctacgacgtt gaccacatcg tcccccaatc tttcttgaag gatgactcca
     8281 tcgacaacaa ggtcttgacc agatccgaca agaacagagg taagtctgac aacgttccat
     8341 ccgaagaggt cgtcaagaag atgaagaact attggagaca gttgttgaac gccaagttga
     8401 tcacccagag aaagttcgac aacttgacta aggccgaaag aggtggtttg tctgaattgg
     8461 acaaggccgg cttcatcaag agacagttgg tcgaaactag acagatcacc aagcacgtcg
     8521 ctcagatctt ggactccaga atgaacacca agtacgacga gaacgacaag ttgattagag
     8581 aggtcaaggt catcaccttg aagtccaagt tggtgtccga cttcagaaag gacttccagt
     8641 tctacaaggt cagagagatc aacaactacc accacgctca tgacgcttac ttgaacgctg
     8701 ttgtcggtac tgccttgatc aagaagtacc caaagttgga atccgagttc gtctacggtg
     8761 actacaaggt ctacgacgtc agaaagatga tcgccaagtc cgaacaagaa atcggtaagg
     8821 ctaccgccaa gtacttcttc tactccaaca tcatgaattt cttcaagacc gagatcacct
     8881 tggccaacgg cgagatcaga aaaagaccat tgatcgaaac taacggcgaa accggtgaaa
     8941 tcgtttggga taagggtaga gacttcgcca ccgtcagaaa ggttttgtct atgccccagg
     9001 tcaacatcgt caagaaaact gaagtccaga ccggcggctt ctccaaagaa tctatcttgc
     9061 caaagagaaa ctcggacaag ttgatcgcca gaaagaagga ctgggaccca aagaaatacg
     9121 gtggtttcga ctctccaacc gtcgcttact ctgttttggt tgtcgctaag gtcgagaagg
     9181 gcaagtctaa gaagttgaag tccgtcaaag agttgttggg catcaccatc atggaaagat
     9241 cgtccttcga gaagaaccca atcgacttct tggaagccaa gggctacaaa gaggtgaaga
     9301 aggacttgat catcaagttg cccaagtact cgttgttcga gttggagaac ggtagaaaga
     9361 gaatgttggc ttccgctggt gaattgcaga agggtaacga attggccttg ccctccaagt
     9421 acgtcaactt cttgtacttg gcctcccact acgaaaagtt gaagggttcc cctgaagata
     9481 acgagcagaa gcagttgttc gtcgagcagc ataagcacta cttggacgaa atcatcgagc
     9541 agatctccga gttctctaag agagtcatct tggctgacgc caacttggac aaggtcttgt
     9601 ctgcttacaa caagcacaga gacaagccaa tcagagagca ggccgaaaac atcatccact
     9661 tgttcacttt gaccaacttg ggtgctccag ctgctttcaa gtacttcgac actaccatcg
     9721 acagaaagag atacacctcc accaaagaag tcttggacgc taccttgatc caccagtcta
     9781 tcactggctt gtacgaaacc agaatcgact tgtctcaatt gggtggtgac gatcctaaga
     9841 agaaaagaaa agttcccggg atgaacccat ctatgaagca gaagcaagaa gagatcaaag
     9901 aaaacatcaa gaactcgtct gtccctagaa gaactttgaa gatgatccaa ccatctgcct
     9961 ctggttcttt ggttggtaga gaaaacgaat tgtctgctgg tttgtctaag agaaagcata
    10021 gaaacgacca cttgacttct actacctctt ctccaggtgt tatcgttcca gaatcttctg
    10081 aaaacaagaa ccttggtggt gtcactcaag aatctttcga cttgatgatc aaagagaacc
    10141 cttcttctca gtactggaaa gaggttgctg aaaagagaag aaaggctttg tgagctgatt
    10201 aatttacgta tattcagttt aatatcaata cgttagctac atttccaatg aacgatacta
    10261 gatattgttt aggattattg aactggtata gataatttta gtgtatattc atgtacttga
    10321 taaatgtaat aatatgtgaa aatgtagttg tacattaact gatagacaac atgctggagt
    10381 atatggcatt aaggttgcta caaagtagaa gcaacctaga cacacctcag aagatagatt
    10441 gggcgagtga tcgtattata tgagaacttc tatgtgatac acttccaagt atgattagtg
    10501 tctacaataa ccttatatta tcggcgcgcc ttcatgaagt acgataaggt tggtaaccga
    10561 ttgactcatt ggttcgtggc ggagaagtac gcagagtaaa accggggccg attcgtggta
    10621 aattctggaa tgattcagag gcgcgacatt tatgcagaca atttgtgttt tgtcgcaaac
    10681 gatgttatag cgaaattttt cactctgtca gataaatgga ttttgtcaaa agggggaagt
    10741 agaaggagaa tgggcccgag atgttctgcc aaattctcag tagcataatg tgaaagaagc
    10801 ccttacattg tccagcctct ggcatcatta aaaaccgtag cggaaaccaa ttgtctctgt
    10861 tcttccctgg cacaccctgg tagccccatc cagttgtagt acatctcaca cgctggcaac
    10921 ttgggacaat cagcaacttt tttttctttt aattttttca gcgcgacatt ttgcctcttc
    10981 tgcgagaaca gactttttca cctccatctc accccccttt gcacttatat aaattggacc
    11041 agttcctccc attgtagaaa aaattttgct ggaccttttt cttttttttt tgtcctttag
    11101 tttcatacaa tctaagtcta tctacaatgt ctaaaggtga agaattattc actggtgttg
    11161 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
    11221 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
    11281 aattgccagt tccatggcca accttagtca ctactttcgg ttatggtgtt caatgttttg
    11341 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
    11401 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
    11461 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
    11521 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
    11581 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
    11641 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
    11701 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
    11761 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
    11821 gtatggatga attgtacaaa taattgatgt aagactttaa ttataagaat acaattgtga
    11881 taaagacaat gtagaagtac tgtagaagtc atggaatagc tgatcaactg tactgttata
    11941 caaaggtact attaacaaaa tagacatata ttttataaca tatataaaca tgtttaaact
    12001 ggagggaatc ctaggataaa aacctccgaa tgacattacc taaaaaaaca cataagtgat
    12061 atctttggaa agcaacagaa gttacaacta ttttatttta ttttacacgt gactgaccgc
    12121 gtgattaggc acgtgactaa tcacgtgctg tataatgaca attggagtga atgtgtaatt
    12181 ttgtgcgata ttttgctgga tggcgcattc gctggaccgg cgagtttgga gttcccgtca
    12241 gctgacggtg cctttgaacg agtcgcaggt tcgtttccaa caagatacat ttatttctct
    12301 tagtgtgttg gccgaaagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
    12361 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcgagtttt tttatcgagc
    12421 atctcttagt gtgttggccg aaaaggtctt ggggtcaggt gaagtcggcg aaacgtgaag
    12481 agaaataata gcgaccgaca gtaaaataat agagagaatc aagattagaa gagaatagga
    12541 gacaaagaag ggtgccttat tcgaatagca ccaattaatt ggtggacaag tgtgtgggtg
    12601 aatttgtata tgcaaaagga gctacacttc tactttttaa ggctcaaaca acttgcggag
    12661 ctcgtggtaa ctctgctctt gtagagtttg atatctctat tttgatctta gtatttgtat
    12721 tgtagctgtc gtatattatt cacttagttg tgagtctcat tatcaagaga actcgagaat
    12781 ggtgggctca tcttcggcct ttttttgcaa gtcaacaatt ctcactcttc caataatacg
    12841 aacacactat ctagaccacc ctaagtcgtc cctatgtcgt atgtttgcct ctactacaaa
    12901 gttactagca aatatccgca gcaacaacag ctgccctctt ccagctggat ccggagccgt
    12961 tgtctgagaa gaataaagag aatatagtcg acacggctgc tgagggtgga gaagagaaga
    13021 aaatcgaaac agacggagat aactaatctg caataaatat tacacttatg taatctaatg
    13081 gtttcttaag aaagtagtcg ttatggagaa gtagtatttt gtattttagt tactttcata
    13141 aatatagcaa gcttatgttg catctcaaac agtgcgaaaa atattttagt ctattttata
    13201 gtatctacaa acaaattttc aattcatcac agactttctt tgtgtactag caacacccta
    13261 aaaacattcg acaatggtca acgtccaaac ctatacacaa agggccgagt ctcaccagtc
    13321 accggtagca cagcgtctct tcaaattaat ggacagcaag aaaactaact tgtgtgcttc
    13381 agtggatgtg aaaacaacct ctgagttctt atctttgatt gataagttgg gtccttacat
    13441 ttgtttggtt aagacccata tcgacattat cgacgatttt tcgtatgaag gtaccattgt
    13501 tcctttgttg gaattgtcca agaagcacaa cttcatgatc tttgaagatc gtaaattcgc
    13561 agacattgga aacactgtca agtctcagta ttcaggtgga gtctacaaga ttgcccaatg
    13621 gtcagatatc accaatgctc acggtgttac tggttctggt attgttcagg gattaaagga
    13681 agctgcaaga gaaactaccg acgaaccaag aggtttattg atgcttgccg aactatcttc
    13741 taagggctcg attgctcatg gtaagtatac tgaagaaact gtggaaatcg ccaaaactga
    13801 caaagagttt gttattgggt tcattgccca aagagacatg ggtggacaag atgaaggttt
    13861 cgattggatt gttatgactc ctggtgtcgg tttggatgac aagggtgaca gtttaggtca
    13921 acaatataga acagtagatg aagttgtttc cacaggaacc gatattatca ttgttggtag
    13981 aggcttattc ggcaagggta gagatcctga tgtagagggc caaagataca gagaagctgg
    14041 atggaacgct tacttgaaga agacaagcca agtgtaaaat atgtaaaatt ttgtaaaaat
    14101 cgtaaaactg taatgtacat cgttaagtat ttacaaacaa ctatatgatt atttacaagc
    14161 agaggcctat tccaaaattt cgaattccag taagtcgtca gtcaatctgg ttctcattat
    14221 ttcaaaaacc aagagggcca cggatgtagc cggtattgtt gtcaatgcat tgcggacaaa
    14281 acctttgtat gccatttgat accaagtcaa ctttgatttt gtcttcatcc taaggatttg
    14341 gttatatgta tcaatgtatg agttgtagta caaggttatg gagggtttgt atgaacgcgt
    14401 agctgatgca ttgtagaaat caagagcttc aagttgaaaa attacgatca acttttactc
    14461 ttttccattg tctcttgtgt atcctcactt tagtttgttt caatcacccc taatactctt
    14521 cacacaatta aaatgactgc taacccttcc ttggtgttga acaagatcga cgacatttcg
    14581 ttcgaaactt acgatgcccc agaaatctct gaacctaccg atgtcctcgt ccaggtcaag
    14641 aaaaccggta tctgtggttc cgacatccac ttctacgccc atggtagaat cggtaacttc
    14701 gttttgacca agccaatggt cttgggtcac gaatccgccg gtactgttgt ccaggttggt
    14761 aagggtgtca cctctcttaa ggttggtgac aacgtcgcta tcgaaccagg tattccatcc
    14821 agattctccg acgaatacaa gagcggtcac tacaacttgt gtcctcacat ggccttcgcc
    14881 gctactccta actccaagga aggcgaacca aacccaccag gtaccttatg taaggcggcc
    14941 gcgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
    15001 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    15061 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    15121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    15181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    15241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    15301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    15361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtt
//
