LOCUS      pEmpty_Control      10604 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Empty control for Cas9 experiments and to clone new guide RNAs for 
            paper.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10604)
  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 10604)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
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: pEmpty_Control
FEATURES             Location/Qualifiers
     source          1..10604
                     /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     3033..3050
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3167..3188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3203..3208,3209..3226,3227..3233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3241..3257
                     /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     3246..3268
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3265..3281
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3265..3281
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3300..3320
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3302..3320
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3338..3736
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3759..3779
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3789..5507
                     /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             6790..6801
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             8319..8339
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8355..8602
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8371..8389)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(8620..8639)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8621..8639)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8649..8666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8649..8665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8658..8680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8806..9261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8893..8912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9103..9124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9392..10195)
                     /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(10196..10411)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(10435..10454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10554..10576
                     /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 ggcggccgcg tttaagagct atgctggaaa cagcatagca
     2641 agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     2701 tgttaactcc gttactgtgg gcttgccata ttttttggaa cttttctgcc ctttttctca
     2761 atgagtaagg agggcgttta tcttttctat tttatttttc tacaaatttg attcatgagt
     2821 ccatggaaca aatatataga actcattttc ttgtttcggt atatatatta actccattga
     2881 agcatttgct tttatcctga ctggctagct agtcgtgcaa tgtatgactt taagccgcgg
     2941 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3001 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3061 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3121 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     3181 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     3241 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     3301 caattaaccc tcactaaagg gaacaaaagc tggagctcat agcttcaaaa tgtttctact
     3361 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
     3421 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
     3481 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
     3541 caataaaaat ttttatcacg tttctttttc ttgaaaattt tttttttgat ttttttctct
     3601 ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
     3661 tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat
     3721 agcaatctaa tctaagtttt ctagagatgt ctagagcccc aaagaagaag agaaaagtta
     3781 gacccgggat ggacaagaag tactccattg ggctcgatat cggcacaaac agcgtcggtt
     3841 gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata
     3901 ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac tccggggaga
     3961 cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc agaaagaatc
     4021 ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat gactctttct
     4081 tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag cgccacccaa
     4141 tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc atatatcatc
     4201 tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc tatctcgcgc
     4261 tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg aacccagaca
     4321 acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag cttttcgaag
     4381 agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct aggctgtcca
     4441 aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag aacggcctgt
     4501 ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct aacttcgacc
     4561 tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat ctcgacaatc
     4621 tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag aacctgtcag
     4681 acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa gctccgctga
     4741 gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg ctgaaggccc
     4801 ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag tctaaaaatg
     4861 gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa tttattaagc
     4921 ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac agagaagatc
     4981 tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt cacctgggcg
     5041 aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa gataacaggg
     5101 aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc ctcgcccggg
     5161 gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact ccctggaact
     5221 tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg atgactaact
     5281 ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact
     5341 tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg agaaagccag
     5401 cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag acgaaccgga
     5461 aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt ttcgactctg
     5521 ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat cacgatctcc
     5581 tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac attcttgagg
     5641 acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa cgcttgaaaa
     5701 cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc cgatatacag
     5761 gatgggggcg gctgtcaaga aaactgatca atggggtatg tgaactgcaa tattaatagc
     5821 acgagaaaat tgagaggaag atagatggga actagtagag ttgatattga tgagatacga
     5881 aaaccacacg taaataaact atccgacgac aagaatagtg cattaaggac ttgattaaga
     5941 tatggtgagc aacgtaatta tcgggctcaa cagtttatta gcaatcgttt tgatagaagc
     6001 gttgatgctg tggaagttgt tctttttact accaacagcc attaacaaat ccattagagt
     6061 gttcgttcgt ttttctgctc agattcaaga aaattattcc atcctcatta tacttttttc
     6121 ttctatttcg tgctccacgt cgaggtatca aggaacatag tttactaaca ttaacgaatt
     6181 catctcctat gaatttactt tttgtataga tccgagacaa gcagagtgga aagacaatcc
     6241 tggattttct taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg
     6301 actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc
     6361 ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga
     6421 ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg
     6481 ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa
     6541 ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc
     6601 cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca
     6661 gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc
     6721 atatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat
     6781 ccgataaaaa tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga
     6841 aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc
     6901 tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc
     6961 agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga
     7021 acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt
     7081 ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca
     7141 attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa
     7201 aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga
     7261 aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca
     7321 gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc
     7381 gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt
     7441 tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag
     7501 tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga
     7561 tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg
     7621 cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg
     7681 tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg
     7741 actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca
     7801 agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc
     7861 tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca
     7921 gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg
     7981 aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag
     8041 tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata
     8101 agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg
     8161 cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa
     8221 aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa
     8281 tcgacctctc tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgt
     8341 gatctcttct cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     8401 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     8461 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     8521 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     8581 acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac
     8641 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     8701 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     8761 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     8821 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     8881 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     8941 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     9001 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     9061 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     9121 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     9181 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     9241 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     9301 ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt tgtgagttta
     9361 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     9421 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     9481 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     9541 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     9601 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     9661 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     9721 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     9781 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     9841 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     9901 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     9961 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
    10021 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
    10081 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
    10141 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
    10201 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
    10261 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
    10321 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
    10381 aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat
    10441 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
    10501 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
    10561 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//