LOCUS       sequence-318637-       10617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Edit Ade2 gene in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10617)
  AUTHORS   Sankaran SM, Smith JD, Roy KR
  TITLE     CRISPR-Cas9 Gene Editing in Yeast: A Molecular Biology and 
            Bioinformatics Laboratory Module for Undergraduate and High School 
            Students.
  JOURNAL   J Microbiol Biol Educ. 2021 May 31;22(2):e00106-21. doi: 
            10.1128/jmbe.00106-21. eCollection 2021 Fall.
  PUBMED    34594460
REFERENCE   2  (bases 1 to 10617)
  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"; doi: 
            "10.1128/jmbe.00106-21"; journalName: "J Microbiol Biol Educ"; date:
            "2021-05-31- 31"; volume: "22"; issue: "2"; pages: "e00106-21"
COMMENT     Sequence Label: pG1
FEATURES             Location/Qualifiers
     source          1..10617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    69..572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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     complement(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3046..3063
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3180..3201
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3216..3221,3222..3239,3240..3246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3254..3270
                     /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)."
     primer_bind     3259..3281
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3278..3294
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3278..3294
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3313..3333
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3315..3333
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3351..3749
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3772..3792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3802..5520
                     /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             6803..6814
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             8332..8352
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8368..8615
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8384..8402)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(8633..8652)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8634..8652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8662..8679)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8662..8678)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8671..8693)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8819..9274
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8906..8925)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9116..9137
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9405..10208)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(10209..10424)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(10448..10467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10567..10589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccactgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gctgcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctctt aattaagtat
     2401 atgtgttatg tagtatactc tttcttcaac aattaaatac tctcggtagc caagttggtt
     2461 taaggcgcaa gactgtaatt tatcactacg aaatcttgag atcgggcgtt cgactcgccc
     2521 ccgggagaga tggccggcat ggtcccagcc tcctcgctgg cgccggctgg gcaacacctt
     2581 cgggtggcga atgggacttt ggcggttgtg gtatatttgg tggtttaaga gctatgctgg
     2641 aaacagcata gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     2701 gtcggtgctt ttttgttaac tccgttactg tgggcttgcc atattttttg gaacttttct
     2761 gccctttttc tcaatgagta aggagggcgt ttatcttttc tattttattt ttctacaaat
     2821 ttgattcatg agtccatgga acaaatatat agaactcatt ttcttgtttc ggtatatata
     2881 ttaactccat tgaagcattt gcttttatcc tgactggcta gctagtcgtg caatgtatga
     2941 ctttaagccg cggtctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3001 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3061 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3121 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3181 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     3241 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     3301 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct catagcttca
     3361 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
     3421 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
     3481 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
     3541 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
     3601 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
     3661 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
     3721 agaaagaaag catagcaatc taatctaagt tttctagaga tgtctagagc cccaaagaag
     3781 aagagaaaag ttagacccgg gatggacaag aagtactcca ttgggctcga tatcggcaca
     3841 aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     3901 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc
     3961 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     4021 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     4081 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     4141 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     4201 accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg
     4261 atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac
     4321 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     4381 cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc
     4441 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     4501 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     4561 tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat
     4621 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     4681 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     4741 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     4801 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     4861 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     4921 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt
     4981 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     5041 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     5101 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     5161 cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     5221 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     5281 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     5341 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     5401 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     5461 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     5521 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     5581 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     5641 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     5701 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg
     5761 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatggggt atgtgaactg
     5821 caatattaat agcacgagaa aattgagagg aagatagatg ggaactagta gagttgatat
     5881 tgatgagata cgaaaaccac acgtaaataa actatccgac gacaagaata gtgcattaag
     5941 gacttgatta agatatggtg agcaacgtaa ttatcgggct caacagttta ttagcaatcg
     6001 ttttgataga agcgttgatg ctgtggaagt tgttcttttt actaccaaca gccattaaca
     6061 aatccattag agtgttcgtt cgtttttctg ctcagattca agaaaattat tccatcctca
     6121 ttatactttt ttcttctatt tcgtgctcca cgtcgaggta tcaaggaaca tagtttacta
     6181 acattaacga attcatctcc tatgaattta ctttttgtat agatccgaga caagcagagt
     6241 ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag
     6301 ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc
     6361 cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag
     6421 ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag
     6481 cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag
     6541 aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc
     6601 cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac
     6661 ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg gctctccgac
     6721 tacgacgtgg atcatatcgt gccccagtct tttctcaaag atgattctat tgataataaa
     6781 gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt
     6841 gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg
     6901 aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc
     6961 ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc
     7021 gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt
     7081 attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg
     7141 agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact
     7201 gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga ctataaagtg
     7261 tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag
     7321 tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga
     7381 gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac
     7441 aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt
     7501 aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac
     7561 agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat
     7621 tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa
     7681 aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa
     7741 aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc
     7801 attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct
     7861 agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc
     7921 ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag
     7981 cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa
     8041 ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat
     8101 aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg
     8161 accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg
     8221 tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc
     8281 tatgaaacaa gaatcgacct ctctcagctc ggtggagaca gcagggctga ccccaagaag
     8341 aagaggaagg tgtgatctct tctcgagtca tgtaattagt tatgtcacgc ttacattcac
     8401 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     8461 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     8521 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     8581 agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag
     8641 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     8701 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     8761 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     8821 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     8881 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     8941 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     9001 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     9061 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     9121 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     9181 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     9241 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     9301 atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat tgaagctcta
     9361 atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg
     9421 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
     9481 tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc
     9541 acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca
     9601 ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca
     9661 ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat
     9721 tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt
     9781 tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca
     9841 ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc
     9901 aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac
     9961 ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata
    10021 tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat
    10081 tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata
    10141 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
    10201 ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat
    10261 actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg
    10321 tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga
    10381 aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca
    10441 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
    10501 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
    10561 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
