LOCUS       Exported               11034 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  p426_Cas9_gRNA-RDS1a without the ribozyme All-in-one CRISPR plasmid 
            for integrating, deleting, or replacing a gene in S. cerevisiae. 
            High-copy, URA3 plasmid contains human codon-optimized Cas9 and a 
            guide RNA targeting RDS1a sequence ATTCAATACGAAATGTGTGC in yeast 
            chromosome 3..
ACCESSION   .
VERSION     .
KEYWORDS    p426_Cas9_gRNA-R-RDS1a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11034)
  AUTHORS   Reider Apel A, d'Espaux L, Wehrs M, Sachs D, Li RA, Tong GJ, Garber 
            M, Nnadi O, Zhuang W, Hillson NJ, Keasling JD, Mukhopadhyay A
  TITLE     A Cas9-based toolkit to program gene expression in Saccharomyces 
            cerevisiae.
  JOURNAL   Nucleic Acids Res. 2017 Jan 9;45(1):496-508. doi: 
            10.1093/nar/gkw1023. Epub 2016 Nov 28.
  PUBMED    27899650
REFERENCE   2  (bases 1 to 11034)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11034
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     71..90
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     324..341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    458..479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        494..524
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    532..548
                     /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     537..559
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     556..572
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     556..572
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     591..611
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        593..611
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        766..841
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        927..1631
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1641..5744
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKNERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIYLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5757..5777
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5793..6040
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6058..6077)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6059..6077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6087..6104)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6087..6103)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6096..6118)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6244..6699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6331..6350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6541..6562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6830..7633)
                     /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(7634..7849)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7873..7892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7992..8014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8052..8070)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8111..9453
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8443..8490
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        9480..9584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9585..10445
                     /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(9803..9822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10616..170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
       61 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      361 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      421 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      481 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      541 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
      601 ctcactaaag ggaacaaaag ctggagctag tatactcttt cttcaacaat taaatactct
      661 cggtagccaa gttggtttaa ggcgcaagac tgtaatttat cactacgaaa tcttgagatc
      721 gggcgttcga ctcgcccccg ggagaattca atacgaaatg tgtgcgtttt agagctagaa
      781 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      841 ctttttttat tttttgtcac tattgttatg taaaatgcca cctctgacag tatggaacgc
      901 aaacttctgt ctagtggata gtcgacatcc ttttgttgtt tccgggtgta caatatggac
      961 ttcctctttt ctggcaacca aacccataca tcgggattcc tataatacct tcgttggtct
     1021 ccctaacatg taggtggcgg aggggagata tacaatagaa cagataccag acaagacata
     1081 atgggctaaa caagactaca ccaattacac tgcctcattg atggtggtac ataacgaact
     1141 aatactgtag ccctagactt gatagccatc atcatatcga agtttcacta ccctttttcc
     1201 atttgccatc tattgaagta ataataggcg catgcaactt cttttctttt tttttctttt
     1261 ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat ggaagacact
     1321 aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca gagctgatga
     1381 ggggtatctc gaagcacacg aaactttttc cttccttcat tcacgcacac tactctctaa
     1441 tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa aaaagtttgc
     1501 tgtcttgcta tcaagtataa atagacctgc aattattaat cttttgtttc ctcgtcattg
     1561 ttctcgttcc ctttcttcct tgtttctttt tctgcacaat atttcaagct ataccaagca
     1621 tacaatcaac ttggaaaaaa atggacaaga agtactccat tgggctcgat atcggcacaa
     1681 acagcgtcgg ttgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag
     1741 ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg
     1801 actccgggga gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc
     1861 gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg
     1921 atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagaacg
     1981 agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa
     2041 ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga
     2101 tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc
     2161 tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc
     2221 agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg
     2281 ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga
     2341 agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat
     2401 ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg
     2461 atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa
     2521 agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca
     2581 aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt
     2641 tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc
     2701 agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca
     2761 aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta
     2821 acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga
     2881 ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga
     2941 aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc
     3001 ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca
     3061 ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa
     3121 ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc
     3181 tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga
     3241 tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca
     3301 agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat
     3361 gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt
     3421 atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg
     3481 acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag
     3541 aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc
     3601 gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc
     3661 agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca
     3721 tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt
     3781 ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca
     3841 aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc
     3901 ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac
     3961 agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc
     4021 aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt
     4081 actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct
     4141 ccgactacga cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata
     4201 ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag
     4261 aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac
     4321 aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag
     4381 ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa
     4441 ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga
     4501 aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata
     4561 aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag
     4621 gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata
     4681 aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg
     4741 ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt acactggcca
     4801 atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt
     4861 gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca
     4921 tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa
     4981 ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat
     5041 tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt
     5101 ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct
     5161 tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc
     5221 tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc
     5281 tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta
     5341 atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc
     5401 agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa
     5461 gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt
     5521 acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc tacttgttta
     5581 ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa
     5641 agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg
     5701 ggctctatga aacaagaatc gacctctctc agctcggtgg agacagcagg gctgacccca
     5761 agaagaagag gaaggtgtga tctcttctcg agtcatgtaa ttagttatgt cacgcttacg
     5821 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     5881 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     5941 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     6001 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc
     6061 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     6121 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6181 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6241 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6301 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6361 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6421 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6481 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6541 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6601 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6661 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     6721 tacgcatctg tgcggtattt cacaccgcat agggtaataa ctgatataat taaattgaag
     6781 ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct
     6841 ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct
     6901 tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag
     6961 agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac
     7021 ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt
     7081 gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag
     7141 tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc
     7201 taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca
     7261 ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt
     7321 actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat
     7381 tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca
     7441 agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca
     7501 gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca
     7561 tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg
     7621 tagctttcga catgatttat cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta
     7681 agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa
     7741 gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc
     7801 aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa aggtggtatg
     7861 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     7921 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     7981 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     8041 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     8101 ttcttagtat gatccaatat caaaggaaat gatagcattg aaggatgaga ctaatccaat
     8161 tgaggagtgg cagcatatag aacagctaaa gggtagtgct gaaggaagca tacgataccc
     8221 cgcatggaat gggataatat cacaggaggt actagactac ctttcatcct acataaatag
     8281 acgcatataa gtacgcattt aagcataaac acgcactatg ccgttcttct catgtatata
     8341 tatatacagg caacacgcag atataggtgc gacgtgaaca gtgagctgta tgtgcgcagc
     8401 tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct ttgaagttcc tattccgaag
     8461 ttcctattct ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg
     8521 aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc
     8581 gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat
     8641 atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct
     8701 ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac
     8761 tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat
     8821 agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta
     8881 tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc
     8941 ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga
     9001 gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac
     9061 ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa
     9121 aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca
     9181 aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg
     9241 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     9301 ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg
     9361 ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt
     9421 ttgttctaca aaatgaagca cagatgcttc gttcaggtgg cacttttcgg ggaaatgtgc
     9481 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     9541 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     9601 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     9661 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     9721 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     9781 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     9841 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     9901 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     9961 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
    10021 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
    10081 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
    10141 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
    10201 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
    10261 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
    10321 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
    10381 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
    10441 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
    10501 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
    10561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
    10621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
    10681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
    10741 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
    10801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    10861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    10921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    10981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaag
//
