LOCUS       sequence-318639-       10615 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 10615)
  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 10615)
  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: pG3
FEATURES             Location/Qualifiers
     source          1..10615
                     /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     3044..3061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3178..3199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3214..3219,3220..3237,3238..3244)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3252..3268
                     /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     3257..3279
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3276..3292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3276..3292
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3311..3331
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3313..3331
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3349..3747
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3770..3790
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3800..5518
                     /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             6801..6812
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             8330..8350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8366..8613
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8382..8400)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(8631..8650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8632..8650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8660..8677)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8660..8676)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8669..8691)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8817..9272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8904..8923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9114..9135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9403..10206)
                     /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(10207..10422)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(10446..10465)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10565..10587
                     /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 gtataatgtc cagagttgtg gtttaagagc tatgctggaa
     2641 acagcatagc aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     2701 cggtgctttt ttgttaactc cgttactgtg ggcttgccat attttttgga acttttctgc
     2761 cctttttctc aatgagtaag gagggcgttt atcttttcta ttttattttt ctacaaattt
     2821 gattcatgag tccatggaac aaatatatag aactcatttt cttgtttcgg tatatatatt
     2881 aactccattg aagcatttgc ttttatcctg actggctagc tagtcgtgca atgtatgact
     2941 ttaagccgcg gtctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3001 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3061 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3121 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3181 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     3241 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3301 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctca tagcttcaaa
     3361 atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca
     3421 cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt
     3481 taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc
     3541 gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga
     3601 tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc
     3661 tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag
     3721 aaagaaagca tagcaatcta atctaagttt tctagagatg tctagagccc caaagaagaa
     3781 gagaaaagtt agacccggga tggacaagaa gtactccatt gggctcgata tcggcacaaa
     3841 cagcgtcggt tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt
     3901 tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga
     3961 ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg
     4021 cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga
     4081 tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga
     4141 gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac
     4201 catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat
     4261 ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct
     4321 gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca
     4381 gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc
     4441 taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa
     4501 gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc
     4561 taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga
     4621 tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa
     4681 gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa
     4741 agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt
     4801 gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca
     4861 gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa
     4921 atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa
     4981 cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat
     5041 tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa
     5101 agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc
     5161 cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac
     5221 tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag
     5281 gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct
     5341 gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat
     5401 gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa
     5461 gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg
     5521 tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta
     5581 tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga
     5641 cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga
     5701 acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg
     5761 ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatggggtat gtgaactgca
     5821 atattaatag cacgagaaaa ttgagaggaa gatagatggg aactagtaga gttgatattg
     5881 atgagatacg aaaaccacac gtaaataaac tatccgacga caagaatagt gcattaagga
     5941 cttgattaag atatggtgag caacgtaatt atcgggctca acagtttatt agcaatcgtt
     6001 ttgatagaag cgttgatgct gtggaagttg ttctttttac taccaacagc cattaacaaa
     6061 tccattagag tgttcgttcg tttttctgct cagattcaag aaaattattc catcctcatt
     6121 atactttttt cttctatttc gtgctccacg tcgaggtatc aaggaacata gtttactaac
     6181 attaacgaat tcatctccta tgaatttact ttttgtatag atccgagaca agcagagtgg
     6241 aaagacaatc ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt
     6301 gatccatgat gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca
     6361 gggggacagt cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg
     6421 aatactgcag accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc
     6481 cgagaatatc gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa
     6541 cagtagggaa aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct
     6601 taaggaacac ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct
     6661 gcagaacggc agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta
     6721 cgacgtggat catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt
     6781 gttgacaaga tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt
     6841 caagaaaatg aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa
     6901 gttcgataat ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt
     6961 catcaaaagg cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga
     7021 ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat
     7081 tactctgaag tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag
     7141 agagatcaac aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc
     7201 acttatcaaa aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta
     7261 cgatgttagg aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta
     7321 cttcttttac agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga
     7381 gattcggaag cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa
     7441 gggtagggat ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa
     7501 aaagaccgaa gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag
     7561 cgacaagctg atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc
     7621 tcctacagtc gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa
     7681 actcaaaagc gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa
     7741 aaaccccatc gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat
     7801 taagcttccc aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag
     7861 tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt
     7921 gtatctggcc agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca
     7981 gctgttcgtg gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt
     8041 ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa
     8101 gcacagggat aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac
     8161 caacttgggc gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta
     8221 cacctctaca aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta
     8281 tgaaacaaga atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa
     8341 gaggaaggtg tgatctcttc tcgagtcatg taattagtta tgtcacgctt acattcacgc
     8401 cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     8461 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     8521 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     8581 aaggttttgg gacgctcgaa ggctttaatt tgcggccggt acccaattcg ccctatagtg
     8641 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     8701 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     8761 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     8821 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     8881 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     8941 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     9001 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     9061 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     9121 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     9181 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     9241 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     9301 ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg aagctctaat
     9361 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca
     9421 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc
     9481 ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac
     9541 atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc
     9601 gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat
     9661 aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc
     9721 tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc
     9781 ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc
     9841 gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa
     9901 tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt
     9961 ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc
    10021 cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc
    10081 cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt
    10141 aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt
    10201 cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac
    10261 tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg
    10321 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa
    10381 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt atggtgcact
    10441 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
    10501 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
    10561 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
