LOCUS       sequence-321817-       11258 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Cas9 and contains natR412 guide which targets the natR 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11258)
  AUTHORS   Parts L, Batte A, Lopes M, Yuen MW, Laver M, San Luis BJ, Yue JX, 
            Pons C, Eray E, Aloy P, Liti G, van Leeuwen J
  TITLE     Natural variants suppress mutations in hundreds of essential genes.
  JOURNAL   Mol Syst Biol. 2021 May;17(5):e10138. doi: 10.15252/msb.202010138.
  PUBMED    34042294
REFERENCE   2  (bases 1 to 11258)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2021-05"; volume: "17(5):e10138. doi"; pages: " 
            10.15252/msb.202010138"
COMMENT     Sequence Label: pML104-natR412
FEATURES             Location/Qualifiers
     source          1..11258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..269
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        290..364
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      369..388
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      complement(506..693)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     614..638
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(721..741)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3136..4854)
                     /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"
     regulatory      4851..4860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4889..5555)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(4996..5019)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(5600..5616)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5601..5617)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5641..5660)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5642..5660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5670..5687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5670..5686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5679..5701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5827..6282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5914..5933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6124..6145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6413..7216)
                     /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(7217..7432)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7456..7475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7575..7597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7635..7653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      7694..9036
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8026..8073
                     /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        9063..9167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9168..9236,9237..10028)
                     /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(9386..9405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10199..10787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10688..10707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10941..10958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11075..11096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11111..11116,11117..11134,11135..11141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11149..11165
                     /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     11154..11176
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11173..11189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11173..11189
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11208..11228
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11210..11228
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt
       61 ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt
      121 agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt
      181 caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga
      241 aacttctccg cagtgaaaga taaatgatcg taccgcacca gtgtcccggg ttttagagct
      301 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      361 ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta attagttata agcatgtgag
      421 ctaagacact gtaattgcca atctaaacga taccacggcc gctctagaga aatggggagc
      481 gatttgcagg catttgctcg gcatgccggt agaggtgtgg tcaataagag cgacctcatg
      541 ctatacctga gaaagcaacc tgacctacag gaaagagtta ctcaagaata agaattttcg
      601 ttttaaaacc taagagtcac tttaaaattt gtatacactt atttttttta taacttattt
      661 aataataaaa atcataaatc ataagaaatt cgcctcgaga gtctaggatc cggaactacc
      721 taccttgcgc tttttcttgg gagctccctc atcccctccg agctgtgaga ggtcaatccg
      781 agtttcgtaa agtccggtga tgctctggtg gatgagggtg gcgtccagca cttctttggt
      841 ggaggtatag cgtttccggt cgatggtggt gtcgaagtac ttgaatgcgg caggggcacc
      901 aagattggtg agggtgaaca ggtggatgat attctcagcc tgctcccgaa ttggtttgtc
      961 tctgtgctta ttgtatgcgc taaggacttt atcgaggttg gcatcagcga gaatgacgcg
     1021 tttagaaaac tcggaaatct gctcaatgat ctcgtccaga taatgcttat gctgctccac
     1081 aaacagttgc ttttgctcgt tgtcctcggg gctccccttc agcttctcgt aatggcttgc
     1141 aaggtagagg aaattcacgt acttggaggg gagagccagc tcattaccct tctgaagttc
     1201 gccagcggag gcgagcatcc gcttccgccc attctccagt tcgaacaggc tatactttgg
     1261 gagcttgatg ataaggtcct tcttgacttc cttgtaacct ttggcttcga gaaagtcaat
     1321 tgggttcttc tcgaaggagc tgcgctccat aatggtgata cccagcagtt ccttcacgct
     1381 cttgagcttc ttagactttc ctttttccac cttagccacg accaggacag aataagccac
     1441 agttggtgaa tcgaatccgc cgtacttctt agggtcccaa tccttcttcc ttgcaatgag
     1501 cttgtcggag ttgcgcttgg ggaggattga ttccttagaa aagcctccgg tttgcacctc
     1561 ggttttcttc acgatgttca cttgaggcat agaaagcact ttgcgcacgg tagcgaaatc
     1621 tctgcccttg tcccacacga tttcgccagt ctccccatta gtctcgatga gcggcctctt
     1681 ccggatctca ccgtttgcaa gggtgatttc ggtcttgaag aaattcatga tgttgctgta
     1741 gaagaagtac tttgcggtgg cctttccgat ctcctgctca gatttggcaa tcatcttgcg
     1801 cacgtcgtag accttgtagt ctccgtacac gaactcgctt tccagctttg ggtacttctt
     1861 gattagggcg gttcccacca cggcattcag atatgcgtca tgagcatgat gatagttatt
     1921 gatctcccgc actttgtaga attgaaagtc ctttctgaaa tcgctgacca gcttggactt
     1981 cagggtaatc accttcacct cgcggataag tttgtcgttt tcgtcgtact tggtattcat
     2041 gcgtgagtca aggatttggg ctacatgttt agtaatctgc cgagtctcca caagttgccg
     2101 tttaatgaat cctgctttgt ccagttcgct cagtccgccc ctctcagcct tagtgagatt
     2161 gtcgaacttc cgctgagtga ttagctttgc gttcagaagc tggcgccaat agttcttcat
     2221 ctttttcacg acttcctctg agggcacgtt atctgacttt ccccggttct tgtctgagcg
     2281 agtcagcacc ttattgtcaa ttgagtcgtc tttgaggaag ctctgaggga cgatgtggtc
     2341 cacgtcgtag tctgagagcc tgttgatatc aagctcttgg tccacataca tatcgcgtcc
     2401 attttgaagg tagtacagat agagcttctc attctgaagc tgggtgtttt ccaccgggtg
     2461 ctctttaagg atctgtgacc cgagctcctt aatcccttcc tcaattcttt tcatccgctc
     2521 cctgctgttc ttctgtccct tctgggtagt ctggttttcc ctggccatct cgatgacaat
     2581 gttctctggt ttatggcgac ccatcacctt gaccagctcg tccacgactt tcacggtctg
     2641 gagaataccc ttcttaatag cgggtgaacc agccagattt gcgatatgct cgtgcagtga
     2701 gtcaccttgt ccagacactt gggccttctg gatgtcctcc ttaaaggtca gggaatcatc
     2761 atggataagt tgcatgaagt tgcgattagc gaatccgtcg ctcttaagga aatcaagaat
     2821 ggtctttccg ctctgtttgt cccggattcc gttgatgagt ttgcggctaa ggcgtcccca
     2881 tccggtatat cttctccgtt taagttgctt catcacttta tcgtcgaaca gatgggcgta
     2941 ggtcttaagc ctctcctcga tcatttcccg gtcctcgaac agagtgagag tcaggacaat
     3001 gtcctcaagg atgtcctctt tctcctcatt gtccagaaaa tccttgtcct tgatgatctt
     3061 aagcagatca tggtaggtac ccaggcttgc gttgaaccga tcttcgaccc cgctaatctc
     3121 cacagagtcg aagcattcaa ttttcttgaa atagtcctcc ttcagctgct tcacggtcac
     3181 ctttctattg gtcttgaaca gaaggtccac aattgccttc ttttgttcgc ctgacaggaa
     3241 tgcgggcttc ctcatccctt cggtgacata tttcaccttg gtcagctcgt tgtacacggt
     3301 aaagtactcg tagagcaggg aatgtttcgg caggaccttc tcattgggga ggttcttatc
     3361 gaagttagtc atccgttcga tgaatgactg agcgctggct cccttgtcca ccacctcttc
     3421 gaagttccag ggagtgatag tttcttctga ctttctggtc atccaagcaa accggctatt
     3481 tcctctggcg aggggtccca cgtagtaggg gatgcggaaa gtgagaattt tctcaatctt
     3541 ctccctgttg tccttaagaa aggggtagaa gtcctcttgg cgccgaagga tggcgtgaag
     3601 ctccccaaga tggatctggt ggggaatgct cccattgtcg aaagtgcgct gcttccgcag
     3661 cagatcctcc cgattcagtt tcacgaggag ctcttcggtg ccgtccatct tctcaaggat
     3721 aggtttgata aacttataaa attcctcttg ggaggcacct ccgtcgatgt agccggcgta
     3781 tccattcttt gactggtcaa agaagatttc cttgtacttc tccggaagct gctgccggac
     3841 gagtgcctta agcagggtca ggtcctggtg gtgctcatca taccgcttaa tcattgaagc
     3901 tgaaagaggg gccttggtga tttcagtgtt cacgcggaga atgtcagaca gcaggatggc
     3961 gtcagaaaga ttcttagcag caaggaaaag gtcggcgtat tggtcaccga tctgggccag
     4021 gaggttgtcg agatcgtcat cataggtgtc cttggacagt tgcagcttgg catcctctgc
     4081 caggtcgaaa ttagacttga aattgggagt cagcccaagt gaaagagcaa tcaggttccc
     4141 gaacagtcca tttttctttt cacccggcag ctgagcgata agattctcaa gtctgcggct
     4201 ctttgacagc cgggctgaca gaatggcctt ggcatcgact ccgcttgcat tgatggggtt
     4261 ctcttcaaag agctggttgt aagtctgcac cagctggatg aacagtttat ccacatcgct
     4321 attgtcagga ttgaggtcgc cctcgatcag gaagtggcct ctgaacttaa tcatgtgggc
     4381 cagggccagg tagataagcc ggaggtctgc cttgtcggtt gagtccacca gctttttgcg
     4441 aagatggtag atggttgggt atttctcgtg ataggccacc tcgtcgacga tattaccgaa
     4501 gatgggatgc cgttcatgct tcttatcctc ttccacgagg aatgattctt ccagcctatg
     4561 gaagaagctg tcgtcgacct ttgccatttc gttggagaat atctcttgaa ggtaacaaat
     4621 ccggttcttc ctcctggtgt accgccgtct agcagtccgc ttgaggcggg ttgcttcagc
     4681 ggtctcccct gaatcaaaga ggagagctcc gatcagattt ttcttgatgg aatgccggtc
     4741 ggtgttcccg agcaccttga acttcttaga gggcaccttg tactcgtcag tgatgacggc
     4801 ccacccgacg ctattagtcc cgatgtccag tccgatagaa tacttcttgt ccatggtggc
     4861 gagatctaat tcgatccata tcagatcttt tgtttgttta tgtgtgttta ttcgaaacta
     4921 agttcttggt gttttaaaac taaaaaaaag actaactata aaagtagaat ttaagaagtt
     4981 taagaaatag atttacagaa ttacaatcaa tacctaccgt ctttatatac ttattagtca
     5041 agtaggggaa taatttcagg gaactggttt caaccttttt tttcagcttt ttccaaatca
     5101 gagagagcag aaggtaatag aaggtgtaag aaaatgagat agatacatgc gtgggtcaat
     5161 tgccttgtgt catcatttac tccaggcagg ttgcatcact ccattgaggt tgtgcccgtt
     5221 ttttgcctgt ttgtgcccct gttctctgta gttgcgctaa gagaatggac ctatgaactg
     5281 atggttggtg aagaaaacaa tattttggtg ctgggattct ttttttttct ggatgccagc
     5341 ttaaaaagcg ggctccatta tatttagtgg atgccaggaa taaactgttc acccagacac
     5401 ctacgatgtt atatattctg tgtaacccgc cccctatttt gggcatgtac gggttacagc
     5461 agaattaaaa ggctaatttt ttgactaaat aaagttagga aaatcactac tattaattat
     5521 ttacgtattc tttgaaatgg cagtattgat aatgataaac tcgaactgac tagtagactg
     5581 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg
     5641 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5701 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5761 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5821 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5881 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5941 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6001 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     6061 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6121 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6181 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6241 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     6301 ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg
     6361 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
     6421 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
     6481 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
     6541 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta
     6601 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
     6661 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
     6721 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
     6781 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
     6841 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
     6901 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa
     6961 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     7021 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     7081 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
     7141 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
     7201 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg
     7261 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
     7321 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
     7381 ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt
     7441 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     7501 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7561 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7621 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     7681 ggtttcttag tatgatccaa tatcaaagga aatgatagca ttgaaggatg agactaatcc
     7741 aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa gcatacgata
     7801 ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat cctacataaa
     7861 tagacgcata taagtacgca tttaagcata aacacgcact atgccgttct tctcatgtat
     7921 atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgc
     7981 agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac gctttgaagt tcctattccg
     8041 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaaaac caaaagcgct
     8101 ctgaagacgc actttcaaaa aaccaaaaac gcaccggact gtaacgagct actaaaatat
     8161 tgcgaatacc gcttccacaa acattgctca aaagtatctc tttgctatat atctctgtgc
     8221 tatatcccta tataacctac ccatccacct ttcgctcctt gaacttgcat ctaaactcga
     8281 cctctacatt ttttatgttt atctctagta ttactcttta gacaaaaaaa ttgtagtaag
     8341 aactattcat agagtgaatc gaaaacaata cgaaaatgta aacatttcct atacgtagta
     8401 tatagagaca aaatagaaga aaccgttcat aattttctga ccaatgaaga atcatcaacg
     8461 ctatcacttt ctgttcacaa agtatgcgca atccacatcg gtatagaata taatcgggga
     8521 tgcctttatc ttgaaaaaat gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt
     8581 ggagtcaggc tttttttatg gaagagaaaa tagacaccaa agtagccttc ttctaacctt
     8641 aacggaccta cagtgcaaaa agttatcaag agactgcatt atagagcgca caaaggagaa
     8701 aaaaagtaat ctaagatgct ttgttagaaa aatagcgctc tcgggatgca tttttgtaga
     8761 acaaaaaaga agtatagatt ctttgttggt aaaatagcgc tctcgcgttg catttctgtt
     8821 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     8881 gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt
     8941 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
     9001 tttttgttct acaaaatgaa gcacagatgc ttcgttcagg tggcactttt cggggaaatg
     9061 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     9121 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     9181 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     9241 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     9301 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     9361 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     9421 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     9481 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     9541 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     9601 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     9661 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     9721 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     9781 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     9841 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     9901 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     9961 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
    10021 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
    10081 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
    10141 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
    10201 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
    10261 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
    10321 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
    10381 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
    10441 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
    10501 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
    10561 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
    10621 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
    10681 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
    10741 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
    10801 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
    10861 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
    10921 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
    10981 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
    11041 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
    11101 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
    11161 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
    11221 actaaaggga acaaaagctg gagctccacc gcggacaa
//
