LOCUS       sequence-462140-       12385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas9 and gRNA GATGAAAACGAAGACGTGAG which targets 
            the TLG1 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12385)
  TITLE     van Leeuwen Lab plasmids 2025
REFERENCE   2  (bases 1 to 12385)
  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"
COMMENT     Sequence Label: pML107-TLG1
FEATURES             Location/Qualifiers
     source          1..12385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     70..86
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     71..87
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        132..798
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     668..691
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      827..836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             833..2551
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             4946..4966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4994..5181
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5049..5073)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(5299..5318)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(5323..5397)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(5418..5686)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     primer_bind     complement(5717..5737)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5717..5735)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5756..5772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5756..5772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5769..5791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5780..5796
                     /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(5804..5810,5811..5828,5829..5834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5849..5870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5987..6004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6158..6746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6238..6257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6917..7708,7709..7777))
                     /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     7540..7559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7778..7882)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7909..9255
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     9296..9314
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9352..9374)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9474..9493
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(9987..11081)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(11082..11489)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(11790..12245)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11927..11948)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12139..12158
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(12371..12385,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(12385,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
       61 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcagt ctactagtca
      121 gttcgagttt atcattatca atactgccat ttcaaagaat acgtaaataa ttaatagtag
      181 tgattttcct aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac
      241 atgcccaaaa tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac
      301 agtttattcc tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa
      361 aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc
      421 attctcttag cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct
      481 caatggagtg atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat
      541 gtatctatct cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag
      601 ctgaaaaaaa aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat
      661 ataaagacgg taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc
      721 tacttttata gttagtcttt tttttagttt taaaacacca agaacttagt ttcgaataaa
      781 cacacataaa caaacaaaag atctgatatg gatcgaatta gatctcgcca ccatggacaa
      841 gaagtattct atcggactgg acatcgggac taatagcgtc gggtgggccg tcatcactga
      901 cgagtacaag gtgccctcta agaagttcaa ggtgctcggg aacaccgacc ggcattccat
      961 caagaaaaat ctgatcggag ctctcctctt tgattcaggg gagaccgctg aagcaacccg
     1021 cctcaagcgg actgctagac ggcggtacac caggaggaag aaccggattt gttaccttca
     1081 agagatattc tccaacgaaa tggcaaaggt cgacgacagc ttcttccata ggctggaaga
     1141 atcattcctc gtggaagagg ataagaagca tgaacggcat cccatcttcg gtaatatcgt
     1201 cgacgaggtg gcctatcacg agaaataccc aaccatctac catcttcgca aaaagctggt
     1261 ggactcaacc gacaaggcag acctccggct tatctacctg gccctggccc acatgattaa
     1321 gttcagaggc cacttcctga tcgagggcga cctcaatcct gacaatagcg atgtggataa
     1381 actgttcatc cagctggtgc agacttacaa ccagctcttt gaagagaacc ccatcaatgc
     1441 aagcggagtc gatgccaagg ccattctgtc agcccggctg tcaaagagcc gcagacttga
     1501 gaatcttatc gctcagctgc cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc
     1561 tctttcactt gggctgactc ccaatttcaa gtctaatttc gacctggcag aggatgccaa
     1621 gctgcaactg tccaaggaca cctatgatga cgatctcgac aacctcctgg cccagatcgg
     1681 tgaccaatac gccgaccttt tccttgctgc taagaatctt tctgacgcca tcctgctgtc
     1741 tgacattctc cgcgtgaaca ctgaaatcac caaggcccct ctttcagctt caatgattaa
     1801 gcggtatgat gagcaccacc aggacctgac cctgcttaag gcactcgtcc ggcagcagct
     1861 tccggagaag tacaaggaaa tcttctttga ccagtcaaag aatggatacg ccggctacat
     1921 cgacggaggt gcctcccaag aggaatttta taagtttatc aaacctatcc ttgagaagat
     1981 ggacggcacc gaagagctcc tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg
     2041 cactttcgac aatgggagca ttccccacca gatccatctt ggggagcttc acgccatcct
     2101 tcggcgccaa gaggacttct acccctttct taaggacaac agggagaaga ttgagaaaat
     2161 tctcactttc cgcatcccct actacgtggg acccctcgcc agaggaaata gccggtttgc
     2221 ttggatgacc agaaagtcag aagaaactat cactccctgg aacttcgaag aggtggtgga
     2281 caagggagcc agcgctcagt cattcatcga acggatgact aacttcgata agaacctccc
     2341 caatgagaag gtcctgccga aacattccct gctctacgag tactttaccg tgtacaacga
     2401 gctgaccaag gtgaaatatg tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga
     2461 acaaaagaag gcaattgtgg accttctgtt caagaccaat agaaaggtga ccgtgaagca
     2521 gctgaaggag gactatttca agaaaattga atgcttcgac tctgtggaga ttagcggggt
     2581 cgaagatcgg ttcaacgcaa gcctgggtac ctaccatgat ctgcttaaga tcatcaagga
     2641 caaggatttt ctggacaatg aggagaaaga ggacatcctt gaggacattg tcctgactct
     2701 cactctgttc gaggaccggg aaatgatcga ggagaggctt aagacctacg cccatctgtt
     2761 cgacgataaa gtgatgaagc aacttaaacg gagaagatat accggatggg gacgccttag
     2821 ccgcaaactc atcaacggaa tccgggacaa acagagcgga aagaccattc ttgatttcct
     2881 taagagcgac ggattcgcta atcgcaactt catgcaactt atccatgatg attccctgac
     2941 ctttaaggag gacatccaga aggcccaagt gtctggacaa ggtgactcac tgcacgagca
     3001 tatcgcaaat ctggctggtt cacccgctat taagaagggt attctccaga ccgtgaaagt
     3061 cgtggacgag ctggtcaagg tgatgggtcg ccataaacca gagaacattg tcatcgagat
     3121 ggccagggaa aaccagacta cccagaaggg acagaagaac agcagggagc ggatgaaaag
     3181 aattgaggaa gggattaagg agctcgggtc acagatcctt aaagagcacc cggtggaaaa
     3241 cacccagctt cagaatgaga agctctatct gtactacctt caaaatggac gcgatatgta
     3301 tgtggaccaa gagcttgata tcaacaggct ctcagactac gacgtggacc acatcgtccc
     3361 tcagagcttc ctcaaagacg actcaattga caataaggtg ctgactcgct cagacaagaa
     3421 ccggggaaag tcagataacg tgccctcaga ggaagtcgtg aaaaagatga agaactattg
     3481 gcgccagctt ctgaacgcaa agctaatcac tcagcggaag ttcgacaatc tcactaaggc
     3541 tgagaggggc ggactgagcg aactggacaa agcaggattc attaaacggc aacttgtgga
     3601 gactcggcag attactaaac atgtagccca aatccttgac tcacgcatga ataccaagta
     3661 cgacgaaaac gacaaactta tccgcgaggt gaaggtgatt accctgaagt ccaagctggt
     3721 cagcgatttc agaaaggact ttcaattcta caaagtgcgg gagatcaata actatcatca
     3781 tgctcatgac gcatatctga atgccgtggt gggaaccgcc ctaatcaaga agtacccaaa
     3841 gctggaaagc gagttcgtgt acggagacta caaggtctac gacgtgcgca agatgattgc
     3901 caaatctgag caggagatcg gaaaggccac cgcaaagtac ttcttctaca gcaacatcat
     3961 gaatttcttc aagaccgaaa tcacccttgc aaacggtgag atccggaaga ggccgctcat
     4021 cgagactaat ggggagactg gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt
     4081 gcgcaaagtg ctttctatgc ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg
     4141 aggcttttct aaggaatcaa tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa
     4201 gaaggattgg gaccctaaga agtacggcgg attcgattca ccaactgtgg cttattctgt
     4261 cctggtcgtg gctaaggtgg aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact
     4321 gctgggtatc accattatgg agcgcagctc cttcgagaag aacccaattg actttctcga
     4381 agccaaaggt tacaaggaag tcaagaagga ccttatcatc aagctcccaa agtatagcct
     4441 gttcgaactg gagaatgggc ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg
     4501 taatgagctg gctctcccct ccaagtacgt gaatttcctc taccttgcaa gccattacga
     4561 gaagctgaag gggagccccg aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa
     4621 gcattatctg gacgagatca ttgagcagat ttccgagttt tctaaacgcg tcattctcgc
     4681 tgatgccaac ctcgataaag tccttagcgc atacaataag cacagagaca aaccaattcg
     4741 ggagcaggct gagaatatca tccacctgtt caccctcacc aatcttggtg cccctgccgc
     4801 attcaagtac ttcgacacca ccatcgaccg gaaacgctat acctccacca aagaagtgct
     4861 ggacgccacc ctcatccacc agagcatcac cggactttac gaaactcgga ttgacctctc
     4921 acagctcgga ggggatgagg gagctcccaa gaaaaagcgc aaggtaggta gttccggatc
     4981 ctagactctc gaggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     5041 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     5101 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct
     5161 tattgaccac acctctaccg gcatgccgag caaatgcctg caaatcgctc cccatttctc
     5221 tagagcggcc gtggtatcgt ttagattggc aattacagtg tcttagctca catgcttata
     5281 actaattaca tgactcgaag acataaaaaa caaaaaaagc accaccgact cggtgccact
     5341 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaacctc
     5401 acgtcttcgt tttcatcgat catttatctt tcactgcgga gaagtttcga acgccgaaac
     5461 atgcgcacca actttcactt ctacagcgtt tgaccaaaat cttttgaaca gaacattgta
     5521 gggtgtgaaa aaatgcgcac ctttaccgct agcccaagag ggcactacaa aatctagagt
     5581 tgtacttcaa acgtacatgt aatcaccttg tatatactcg aaagaaaaca tcaagtttct
     5641 gtataaatat gagtgaaagc ataatcatac attatctttt caaagattgt ccgcggtgga
     5701 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     5761 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5821 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5881 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa cgaagcatct
     7921 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     7981 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     8041 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     8101 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     8161 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     8221 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     8281 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     8341 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     8401 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     8461 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     8521 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     8581 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     8641 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     8701 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     8761 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     8821 ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     8881 atactttcta gctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac
     8941 gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc
     9001 tatatctgcg tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta
     9061 tgcttaaatg cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc
     9121 ctgtgatatt atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag
     9181 ctgttctata tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc
     9241 ctttgatatt ggatcatact aagaaaccat tattatcatg acattaacct ataaaaatag
     9301 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     9361 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     9421 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9481 agagcagatt gtactgagag tgcaccatat cgactacgtc gttaaggccg tttctgacag
     9541 agtaaaattc ttgagggaac tttcaccatt atgggaaatg gttcaagaag gtattgactt
     9601 aaactccatc aaatggtcag gtcattgagt gttttttatt tgttgtattt tttttttttt
     9661 agagaaaatc ctccaatata taaattagga atcatagttt catgattttc tgttacacct
     9721 aactttttgt gtggtgccct cctccttgtc aatattaatg ttaaagtgca attctttttc
     9781 cttatcacgt tgagccatta gtatcaattt gcttacctgt attcctttac atcctccttt
     9841 ttctccttct tgataaatgt atgtagattg cgtatatagt ttcgtctacc ctatgaacat
     9901 attccatttt gtaatttcgt gtcgtttcta ttatgaattt catttataaa gtttatgtac
     9961 aaatatcata aaaaaagaga atctttttaa gcaaggattt tcttaacttc ttcggcgaca
    10021 gcatcaccga cttcggtggt actgttggaa ccacctaaat caccagttct gatacctgca
    10081 tccaaaacct ttttaactgc atcttcaatg gccttacctt cttcaggcaa gttcaatgac
    10141 aatttcaaca tcattgcagc agacaagata gtggcgatag ggttgacctt attctttggc
    10201 aaatctggag cagaaccgtg gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc
    10261 aaagaggcca aggacgcaga tggcaacaaa cccaaggaac ctgggataac ggaggcttca
    10321 tcggagatga tatcaccaaa catgttgctg gtgattataa taccatttag gtgggttggg
    10381 ttcttaacta ggatcatggc ggcagaatca atcaattgat gttgaacctt caatgtagga
    10441 aattcgttct tgatggtttc ctccacagtt tttctccata atcttgaaga ggccaaaaca
    10501 ttagctttat ccaaggacca aataggcaat ggtggctcat gttgtagggc catgaaagcg
    10561 gccattcttg tgattctttg cacttctgga acggtgtatt gttcactatc ccaagcgaca
    10621 ccatcaccat cgtcttcctt tctcttacca aagtaaatac ctcccactaa ttctctgaca
    10681 acaacgaagt cagtaccttt agcaaattgt ggcttgattg gagataagtc taaaagagag
    10741 tcggatgcaa agttacatgg tcttaagttg gcgtacaatt gaagttcttt acggattttt
    10801 agtaaacctt gttcaggtct aacactacct gtaccccatt taggaccacc cacagcacct
    10861 aacaaaacgg catcagcctt cttggaggct tccagcgcct catctggaag tggaacacct
    10921 gtagcatcta tagcagcacc accaattaaa tgattttcga aatcgaactt gacattggaa
    10981 cgaacatcag aaatagcttt aagaacctta atggcttcgg ctgtgatttc ttgaccaacg
    11041 tggtcacctg gcaaaacgac gatcttctta ggggcagaca ttagaatggt atatccttga
    11101 aatatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat
    11161 tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat
    11221 tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc
    11281 gctctcacct ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga
    11341 cctctgaaat taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct
    11401 gtgtgttctc gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca
    11461 tcttacgata cctgagtatt cccacagtta actgcggtca agatatttct tgaatcaggc
    11521 gccttagacc gctcggccaa acaaccaatt acttgttgag aaatagagta taattatcct
    11581 ataaatataa cgtttttgaa cacacatgaa caaggaagta caggacaatt gattttgaag
    11641 agaatgtgga ttttgatgta attgttggga ttccattttt aataaggcaa taatattagg
    11701 tatgtagata tactagaagt tctcctcgac cggtcgatat gcggtgtgaa ataccgcaca
    11761 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
    11821 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
    11881 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
    11941 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
    12001 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
    12061 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
    12121 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
    12181 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
    12241 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
    12301 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
    12361 ccagggtttt cccagtcacg acgtt
//
