LOCUS       sequence-322090-       10928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression of SpCas9 in yeast after assembly with PID 
            fragments: TEF1p-SpCas9::KanR-1xNLS-CYC1t (KanR cassette in place of
            the PID).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10928)
  AUTHORS   Goldberg GW, Spencer JM, Giganti DO, Camellato BR, Agmon N, Ichikawa
            DM, Boeke JD, Noyes MB
  TITLE     Engineered dual selection for directed evolution of SpCas9 PAM 
            specificity.
  JOURNAL   Nat Commun. 2021 Jan 13;12(1):349. doi: 10.1038/s41467-020-20650-x.
  PUBMED    33441553
REFERENCE   2  (bases 1 to 10928)
  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"; journalName: "Nat 
            Commun."; date: "2021-01-13"; volume: "12(1):349. doi"; pages: " 
            10.1038/s41467-020-20650-x"
COMMENT     Sequence Label: pGG431
FEATURES             Location/Qualifiers
     source          1..10928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    16..519
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        556..660
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(661..729,730..1521)
                     /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(879..898)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1692..2280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2181..2200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2434..2451
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2604..2609,2610..2627,2628..2634)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2642..2658
                     /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     2647..2669
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2666..2682
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2666..2682
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2701..2721
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2703..2721
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2739..3137
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3141..3157
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3141..3157
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3169..4887
                     /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             5756..5767
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(6472..7266)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6583..6602)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7193..7212
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             7514..7534
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7550..7797
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7566..7584)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7815..7834)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7816..7834)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7844..7860)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      8006..8461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8093..8112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8303..8324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8761..9165
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             9178..10272
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(10765..10784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10884..10902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tggtttctta ggacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat
       61 ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta
      121 gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt
      181 taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat
      241 taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg
      301 tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc
      361 aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca
      421 aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat atatttatat
      481 taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact
      541 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      601 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      661 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      721 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      781 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      841 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      901 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      961 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1021 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1081 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1141 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1201 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1261 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1321 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggatct
     1381 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1441 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1501 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1561 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1621 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1681 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1741 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1801 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1861 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1921 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1981 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2041 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2101 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2161 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2221 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2281 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2341 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2401 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2461 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2521 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttac
     2581 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcctat gttgtgtgga
     2641 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     2701 gcaattaacc ctcactaaag ggaacaaaag ctggagctca tagcttcaaa atgtttctac
     2761 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     2821 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     2881 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     2941 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga tttttttctc
     3001 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
     3061 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
     3121 tagcaatcta atctaagttt tctagaacta gtggatcccc cgggaaaaat ggacaagaag
     3181 tactccattg ggctcgatat cggcacaaac agcgtcggtt gggccgtcat tacggacgag
     3241 tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag
     3301 aagaacctca ttggcgccct cctgttcgac tccggggaga cggccgaagc cacgcggctc
     3361 aaaagaacag cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag
     3421 atctttagta atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc
     3481 tttttggtgg aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac
     3541 gaggtggcgt accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac
     3601 agcactgata aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt
     3661 cggggacact tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc
     3721 tttatccaac tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc
     3781 ggagttgacg ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac
     3841 ctcatcgcac agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg
     3901 tcactcgggc tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt
     3961 caactgagca aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac
     4021 cagtacgcag accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat
     4081 attctgcgag tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc
     4141 tatgatgagc accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct
     4201 gagaagtaca aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac
     4261 ggcggagcaa gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac
     4321 ggcaccgagg agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact
     4381 ttcgacaatg gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg
     4441 cggcaagagg atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc
     4501 acatttcgga taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg
     4561 atgactcgca aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag
     4621 ggggcctctg cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac
     4681 gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc
     4741 accaaggtca aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag
     4801 aagaaagcta tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc
     4861 aaagaagact atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag
     4921 gatcgcttca acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag
     4981 gacttcctgg acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg
     5041 ttgtttgaag atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac
     5101 gacaaagtca tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga
     5161 aaactgatca atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag
     5221 tccgatggat ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt
     5281 aaggaggaca tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc
     5341 gctaatcttg caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg
     5401 gatgaactcg tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc
     5461 cgagagaacc aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt
     5521 gaagagggta taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc
     5581 cagcttcaga atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg
     5641 gatcaggaac tggacatcaa tcggctctcc gactacgacg tggatcatat cgtgccccag
     5701 tcttttctca aagatgattc tattgataat aaagtgttga caagatccga taaaaataga
     5761 gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg
     5821 cagctgctga acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa
     5881 cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca
     5941 cgccagatca ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat
     6001 gaaaatgaca aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca
     6061 gatttcagaa aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg
     6121 catgatgcct acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt
     6181 gaatctgaat ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag
     6241 tctgagcagg aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat
     6301 tttttcaaga ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa
     6361 acaaacggag aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg
     6421 aaggtcctgt ccatgccgca agtcagctag cctcgaaccc cagagtcccg ctcagaagaa
     6481 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     6541 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     6601 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     6661 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     6721 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     6781 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     6841 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     6901 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     6961 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     7021 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     7081 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     7141 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     7201 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     7261 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     7321 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     7381 gggcgcccca gctggcaatt ccgacgtcac ccgtaggcat caaataaaac gaaaggctca
     7441 gtcgagggcc caatatgggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga
     7501 cagcagggct gaccccaaga agaagaggaa ggtgtgatct cttctcgagt catgtaatta
     7561 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     7621 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     7681 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     7741 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     7801 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     7861 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     7921 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     7981 cgcagcctga atggcgaatg gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8041 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8101 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8161 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8221 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8281 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     8341 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     8401 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     8461 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatcgacgg
     8521 tcgaggagaa cttctagtat atccacatac ctaatattat tgccttatta aaaatggaat
     8581 cccaacaatt acatcaaaat ccacattctc ttcaaaatca attgtcctgt acttccttgt
     8641 tcatgtgtgt tcaaaaacgt tatatttata ggataattat actctatttc tcaacaagta
     8701 attggttgtt tggccgagcg gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt
     8761 aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta
     8821 tttttttcct caacataacg agaacacaca ggggcgctat cgcacagaat caaattcgat
     8881 gactggaaat tttttgttaa tttcagaggt cgcctgacgc atataccttt ttcaactgaa
     8941 aaattgggag aaaaaggaaa ggtgagaggc cggaaccggc ttttcatata gaatagagaa
     9001 gcgttcatga ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc
     9061 tattgttttt tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt
     9121 acatttcagc aatatatata tatatttcaa ggatatacca ttctaatgtc tgcccctatg
     9181 tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc
     9241 gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa gttcgatttc
     9301 gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgtcccact tccagatgag
     9361 gcgctggaag cctccaagaa ggttgatgcc gttttgttag gtgctgtggg tggtcctaaa
     9421 tggggtaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa agaacttcaa
     9481 ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga cttatctcca
     9541 atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt agtgggaggt
     9601 atttactttg gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa
     9661 tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc cctacaacat
     9721 gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc ttcaagatta
     9781 tggagaaaaa ctgtggagga aaccatcaag aacgaattcc ctacattgaa ggttcaacat
     9841 caattgattg attctgccgc catgatccta gttaagaacc caacccacct aaatggtatt
     9901 ataatcacca gcaacatgtt tggtgatatc atctccgatg aagcctccgt tatcccaggt
     9961 tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa caccgcattt
    10021 ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa ggttgaccct
    10081 atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa
    10141 ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag aactggtgat
    10201 ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa
    10261 atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt tataaatgaa
    10321 attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg tagacgaaac
    10381 tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatag taaaggaata
    10441 caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac
    10501 attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga
    10561 aactacgatt cctaatttga tattggagga ttttctctaa aaaaaaaaaa atacaacaaa
    10621 taaaaaacac tcaatgacct gaccatttga tggagtttaa gtcaatacct tcttgaacca
    10681 tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg gccttacgac
    10741 gtagtcgata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
    10801 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
    10861 ttacagacaa gctgtgacaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
    10921 gataataa
//
