LOCUS       sequence-202306-       11542 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Single copy yeast plasmid expressing Cpf1 from Francisella novicida 
            U112 (FnCpf1), codon optimized for expression in Saccharomyces 
            cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11542)
  AUTHORS   Verwaal R, Buiting-Wiessenhaan N, Dalhuijsen S, Roubos JA
  TITLE     CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2017 Sep 8. doi: 10.1002/yea.3278.
  PUBMED    28886218
REFERENCE   2  (bases 1 to 11542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast. 2017
            Sep 8. doi: 10.1002/yea.3278."
COMMENT     Sequence Label: pCSN068
FEATURES             Location/Qualifiers
     source          1..11542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     494..512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(550..572)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     672..691
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        712..992
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             993..1667
                     /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(1770..2225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1907..1928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2119..2138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2351..2373
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2365..2382
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2366..2382
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2392..2411
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2392..2410
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7337..7357
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    7706..7739
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            7785..9141
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        7785..8128
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             8129..8938
                     /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(8196..8215)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      8944..9141
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    9189..9222
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(9290..9310)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9290..9308)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9329..9345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9329..9345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9342..9364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9353..9369
                     /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(9377..9383,9384..9401,9402..9407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9422..9443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9560..9577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9731..10319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9811..9830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10490..11281,11282..11350))
                     /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     11113..11132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11351..11455)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(11492..11542,1..453)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag
       61 atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct
      121 ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc
      181 ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct
      241 tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta
      301 tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat
      361 acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga
      421 tacgaggcgc gtgtaagtta caggcaagcg atccgtctaa gaaaccatta ttatcatgac
      481 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
      541 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
      601 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
      661 gcttaactat gcggcatcag agcagattgt actgagagtg caccatagat cacgacatta
      721 ctatatatat aatataggaa gcatttaata gaacagcatc gtaatatatg tgtactttgc
      781 agttatgacg ccagatggca gtagtggaag atattcttta ttgaaaaata gcttgtcacc
      841 ttacgtacaa tcttgatccg gagcttttct ttttttgccg attaagaatt aattcggtcg
      901 aaaaaagaaa aggagagggc caagagggag ggcattggtg actattgagc acgtgagtat
      961 acgtgattaa gcacacaaag gcagcttgga gtatgtctgt tattaatttc acaggtagtt
     1021 ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgctctag
     1081 attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc
     1141 cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc
     1201 cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca
     1261 atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag
     1321 agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac
     1381 tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg
     1441 ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc
     1501 ccgaaagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc
     1561 ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa
     1621 aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaataggtt attactgagt
     1681 agtatttatt taagtattgt ttgtgcactt gccgatctat gcggtgtgaa ataccgcaca
     1741 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     1801 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1861 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1921 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1981 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2041 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2101 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2161 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2221 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2281 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     2341 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
     2401 tcactatagg gcgaattggg taccttttct ttttttgcgg tcacccccat gtggcgggga
     2461 ggcagaggag taggtagagc aacgaatcct actatttatc caaattagtc taggaactct
     2521 ttttctagat tttttagatt tgagggcaag cgctgttaac gactcagaaa tgtaagcact
     2581 acggagtaga acgagaaatc cgccataggt ggaaatccta gcaaaatctt gcttacccta
     2641 gctagcctca ggtaagctag ccttagcctg tcaaattttt ttcaaaattt ggtaagtttc
     2701 tactagcaaa gcaaacacgg ttcaacaaac cgaaaactcc actcattata cgtggaaacc
     2761 gaaacaaaaa aacaaaaacc aaaatactcg ccaatgagaa agttgctgcg tttctacttt
     2821 cgaggaagag gaactgagag gattgactac gaaaggggca aaaacgagtc gtattctccc
     2881 attattgtct gctaccacgc ggtctagtag aataagcaac cagtcaacgc taagacaggt
     2941 aatcaaaata ccagtctgct ggctacgggc tagtttttac ctcttttaga acccactgta
     3001 aaagtccgtt gtaaagcccg ttctcactgt tggcgttttt ttttttttgg tttagtttct
     3061 tatttttcat ttttttcttt catgaccaaa aacaaacaaa tctcgcgatt tgtactgcgg
     3121 ccactggggc gtggccaaaa aaatgacaaa tttagaaacc ttagtttctg atttttcctg
     3181 ttatgaggag atatgataaa aaatattact gctttattgt ttttttttta tctactgaaa
     3241 tagagaaact tacccaagga ggaggcaaaa aaaagagtat atatacagca gctaccattc
     3301 agattttaat atattctttt ctcttcttct acactattat tataataatt ttactatatt
     3361 catttttagc ttaaaacctc atagaatatt attcttcagt cactcgctta aatacttatc
     3421 aaaaatgtcc atctaccaag agtttgtcaa caaatactct ttgtctaaga ctttacgttt
     3481 cgaattgatt ccacaaggta agactttgga aaacattaag gctcgtggtt tgatcttgga
     3541 cgacgaaaag agagccaagg attacaagaa ggccaagcaa atcatcgata agtaccacca
     3601 attcttcatt gaagaaatct tatcctctgt ctgtatctcc gaagatctat tgcaaaacta
     3661 ctccgatgtc tacttcaagt tgaaaaagtc tgacgatgac aacttgcaaa aggatttcaa
     3721 atctgccaag gacaccatca agaaacaaat ttctgaatac ataaaggact ctgaaaaatt
     3781 taagaactta ttcaaccaaa acttgatcga tgctaagaag ggtcaagaat ctgacttgat
     3841 cttgtggttg aagcaatcta aggacaacgg tatcgaattg ttcaaggcta actctgatat
     3901 cactgacatt gacgaagctt tggaaatcat caagtctttc aaaggttgga ctacctattt
     3961 caagggtttc cacgaaaacc gtaagaatgt ctactcttcc aacgacatcc caacttccat
     4021 catttacaga atcgttgacg acaatttgcc aaagttccta gaaaacaagg ccaaatacga
     4081 atccttgaag gacaaggctc cagaagccat taactacgaa caaatcaaga aggacttggc
     4141 tgaagaatta actttcgaca ttgactacaa gacttctgaa gttaaccaaa gagttttctc
     4201 tttggacgaa gtcttcgaga ttgctaactt caacaactac ttgaaccaat ctggtatcac
     4261 caaattcaac accatcatcg gtggtaagtt cgtcaacggt gaaaacacca agagaaaggg
     4321 tatcaacgaa tacattaact tgtactctca gcaaatcaac gacaagactt taaagaaata
     4381 caagatgtct gttttgttca agcaaatttt gtctgacact gaatccaagt cttttgtcat
     4441 tgataagttg gaagatgatt ccgacgtcgt taccaccatg caatctttct acgagcaaat
     4501 cgctgctttc aagaccgttg aagaaaagtc tattaaggaa actttgtctt tgttgttcga
     4561 cgatttgaag gctcaaaagt tggatttgtc caagatttac ttcaagaatg acaagtcttt
     4621 gactgatttg tctcaacaag ttttcgatga ctactccgtt attggtactg ctgtcttgga
     4681 atacatcacc caacaaatcg ctcctaagaa cttggacaac ccatccaaga aggaacaaga
     4741 attgattgcc aagaagaccg aaaaagctaa atacttgtct ttggaaacca ttaaattggc
     4801 tttagaagag ttcaacaagc acagagatat tgacaagcaa tgtagattcg aagaaatttt
     4861 ggctaacttc gctgctatcc caatgatctt cgacgaaatt gctcaaaaca aggataactt
     4921 ggctcaaatc tccatcaagt accaaaacca gggtaagaag gatttgttgc aagcctccgc
     4981 tgaagatgac gtcaaggcca tcaaagattt attggaccaa actaacaact tgttgcacaa
     5041 gctaaagatc ttccacatct ctcaatctga agataaagct aacattttgg ataaggacga
     5101 acacttctac ttagttttcg aagaatgtta cttcgaacta gctaacatcg tcccattgta
     5161 caacaagatc agaaactaca ttactcaaaa accatactct gatgaaaagt tcaagttaaa
     5221 cttcgaaaat tctaccttgg ctaacggttg ggacaagaac aaagaaccag acaacaccgc
     5281 catcttgttc attaaggacg acaagtacta cttgggtgtc atgaacaaaa agaacaataa
     5341 gattttcgac gacaaagcta tcaaggagaa caagggtgaa gggtacaaga agattgttta
     5401 taagttgttg ccaggtgcta acaaaatgtt gccaaaggtt ttcttctccg ctaagtctat
     5461 caagttctat aacccttctg aagacatttt gagaatcaga aaccactcca cccacaccaa
     5521 gaacggttct ccacaaaagg gttacgaaaa gttcgaattc aacatcgaag actgtagaaa
     5581 gttcatcgat ttctacaagc aatccatttc caagcatcca gaatggaagg atttcggttt
     5641 cagattctct gacactcaaa gatacaactc cattgatgaa ttctacagag aagtcgaaaa
     5701 ccaaggttac aagttgactt tcgaaaacat ctctgaatct tacattgatt ccgtcgttaa
     5761 ccaaggtaag ttgtacttgt tccaaatcta caacaaggac ttctccgcct actccaaggg
     5821 tagaccaaac ttgcacacct tgtactggaa ggctttgttt gacgaaagaa acttgcaaga
     5881 tgttgtctac aagctgaacg gtgaagctga attgttttac agaaagcaaa gtattccaaa
     5941 gaaaatcact cacccagcta aggaagccat tgccaacaag aataaagaca accctaagaa
     6001 ggaatctgtt tttgaatacg acttaatcaa ggataagaga ttcaccgaag acaaattctt
     6061 cttccattgt ccaatcacca tcaacttcaa gtcctctgga gctaacaagt tcaacgatga
     6121 aatcaacttg ttattgaagg aaaaggctaa cgatgttcac atcttgtcta tcgatcgtgg
     6181 tgaaagacac ttggcttact acactttggt tgatggtaag ggtaacatca ttaagcaaga
     6241 cacctttaac attatcggta acgacagaat gaagaccaac taccacgaca aattggctgc
     6301 tattgaaaag gacagagact ctgctagaaa ggactggaaa aagatcaata acatcaagga
     6361 aatgaaggaa ggttacttgt ctcaagttgt ccatgaaatt gctaagttgg ttatcgaata
     6421 caatgctatc gttgtcttcg aggatttgaa cttcggtttt aagagaggta gattcaaggt
     6481 tgaaaagcaa gtttaccaaa aattggaaaa gatgttgatt gaaaagttga actacttagt
     6541 cttcaaggac aatgaatttg acaagactgg tggtgtcttg agagcttacc aattgactgc
     6601 tccattcgaa actttcaaga agatgggtaa gcaaaccggt atcatctact acgttccagc
     6661 tggtttcact tctaaaatct gtccagttac cggtttcgtc aaccaattgt acccaaagta
     6721 cgaatccgtt tccaagtccc aagaattttt ctccaagttc gacaagatct gttacaactt
     6781 agacaagggt tatttcgagt tttccttcga ttacaaaaac tttggtgaca aagccgctaa
     6841 gggtaaatgg actatcgctt ctttcggttc tagattgatc aacttccgta actccgataa
     6901 gaaccacaac tgggacacta gagaagttta cccaaccaag gaattagaaa aattgttgaa
     6961 ggactactct attgaatacg gtcacggtga atgtatcaag gctgccatct gtggtgaatc
     7021 tgataagaag ttcttcgcta agctaacttc cgtcttgaac accattttgc aaatgagaaa
     7081 ctccaagacc ggtactgaat tggactactt gatttctcca gttgccgatg ttaacggtaa
     7141 cttcttcgac tctagacaag ctccaaagaa catgccacaa gacgctgatg ctaacggtgc
     7201 ctaccacatt ggtttgaagg gtttgatgtt gttgggtcgt attaagaaca accaagaagg
     7261 taagaagttg aacctagtca ttaagaacga agaatacttc gaatttgttc aaaacagaaa
     7321 caactccaga gctgacccaa agaagaagag aaaagtctaa aggagttaaa ggcaaagttt
     7381 tcttttctag agccgttccc acaaataatt atacgtatat gcttcttttc gtttactata
     7441 tatctatatt tacaagcctt tattcactga tgcaatttgt ttccaaatac ttttttggag
     7501 atctcataac tagatatcat gatggcgcaa cttggcgcta tcttaattac tctggctgcc
     7561 aggcccgtgt agagggccgc aagaccttct gtacgccata tagtctctaa gaacttgaac
     7621 aagtttctag acctattgcc gcctttcgga tcgctattgt tgcggccgcc agctgaagct
     7681 tcgtacgctg caggtcgacg aattctaccg ttcgtataat gtatgctata cgaagttata
     7741 gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
     7801 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     7861 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     7921 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     7981 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     8041 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     8101 ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc
     8161 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     8221 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     8281 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     8341 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     8401 tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata
     8461 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     8521 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     8581 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     8641 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     8701 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     8761 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     8821 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     8881 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     8941 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     9001 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     9061 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     9121 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa
     9181 acgagctcat aacttcgtat aatgtatgct atacgaacgg tagaattcga tatcagatcc
     9241 actagtggcc tatgcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     9301 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     9361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     9421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     9481 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     9541 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     9601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     9661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     9721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     9781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     9841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     9901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     9961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
    10021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
    10081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
    10141 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
    10201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
    10261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
    10321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
    10381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
    10441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    10501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    10561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    10621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    10681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    10741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    10801 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    10861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    10921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    10981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    11041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    11101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    11161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    11221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    11281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    11341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    11401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    11461 ttccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt
    11521 aaatttttta atataaatat at
//
