LOCUS       sequence-330958-       11357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression of SpCas9-Zif268 in yeast after assembly with 
            PID fragments: 
            TEF1p-SpCas9::KanR-1xNLS-3xHA-1xNLS-Zif268-1xNLS-CYC1t (KanR 
            cassette in place of the PID).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11357)
  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 11357)
  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: pGG442
FEATURES             Location/Qualifiers
     source          1..11357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     403..419
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     403..419
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             431..2149
                     /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             3018..3029
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3734..4528)
                     /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(3845..3864)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4455..4474
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             4776..4796
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4797..4886
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4899..4925
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             5217..5237
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5253..5500
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5269..5287)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5518..5537)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5519..5537)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5547..5564)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5547..5563)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5556..5578)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5704..6159
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5791..5810)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6001..6022
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6460..6867
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6868..7962
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(8456..8475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8575..8593)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    8635..9138
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        9175..9279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9280..9348,9349..10140)
                     /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(9498..9517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10311..10899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10800..10819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11053..11070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11187..11208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11223..11228,11229..11246,11247..11253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11261..11277
                     /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     11266..11288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11285..11301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11285..11301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11320..11340
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11322..11340
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 catagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc
       61 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct
      121 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt
      181 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa
      241 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
      301 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
      361 cttgctcatt agaaagaaag catagcaatc taatctaagt tttctagaac tagtggatcc
      421 cccgggaaaa atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg
      481 ttgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
      541 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
      601 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
      661 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
      721 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
      781 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
      841 tctgaggaag aagcttgtag acagcactga taaggctgac ttgcggttga tctatctcgc
      901 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
      961 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     1021 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     1081 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
     1141 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
     1201 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
     1261 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
     1321 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
     1381 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
     1441 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
     1501 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
     1561 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
     1621 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
     1681 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
     1741 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
     1801 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
     1861 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
     1921 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
     1981 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
     2041 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
     2101 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
     2161 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
     2221 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
     2281 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
     2341 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
     2401 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
     2461 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
     2521 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
     2581 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
     2641 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
     2701 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
     2761 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
     2821 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
     2881 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
     2941 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
     3001 gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa
     3061 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
     3121 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
     3181 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
     3241 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
     3301 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
     3361 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
     3421 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
     3481 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
     3541 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
     3601 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
     3661 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caagtcagct agcctcgaac
     3721 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3781 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     3841 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     3901 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     3961 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     4021 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     4081 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     4141 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     4201 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     4261 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     4321 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     4381 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     4441 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4501 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     4561 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     4621 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc acccgtaggc
     4681 atcaaataaa acgaaaggct cagtcgaggg cccaatatgg gctctatgaa acaagaatcg
     4741 acctctctca gctcggtgga gacggcaccg gcggacctaa gaagaaaagg aaggtctacc
     4801 catacgatgt tcctgactat gcgggctatc cctatgacgt cccggactat gcaggatcgt
     4861 atccttatga cgttccagat tacgctggat ccgccgctcc ggcagctaag aaaaagaaac
     4921 tggatttcga atccggacgc ccatatgctt gccctgtcga gtcctgcgat cgccgctttt
     4981 ctcgctcgga tgagcttacc cgccatatcc gcatccatac cggtcagaag cccttccagt
     5041 gtcgaatctg catgcgtaac ttcagtcgta gtgaccacct taccacccac atccgcaccc
     5101 acacaggcga gaagcctttt gcctgtgaca tttgtgggag gaagtttgcc aggagtgatg
     5161 aacgcaagag gcataccaaa atccatttaa gacagaagga cgtcagcagg gctgacccca
     5221 agaagaagag gaaggtgtga tctcttctcg agtcatgtaa ttagttatgt cacgcttaca
     5281 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     5341 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     5401 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     5461 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc
     5521 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     5581 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5641 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5701 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6121 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     6181 tacgcatctg tgcggtattt cacaccgcat atcgaccggt cgaggagaac ttctagtata
     6241 tctacatacc taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc
     6301 cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt
     6361 atatttatag gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg
     6421 tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta
     6481 tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc aacataacga
     6541 gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat
     6601 ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag
     6661 gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt
     6721 gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt
     6781 ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata
     6841 tatatatttc aaggatatac cattctaatg tctgccccta agaagatcgt cgttttgcca
     6901 ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa agctatttct
     6961 gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg tgctgctatc
     7021 gatgctacag gtgttccact tccagatgag gcgctggaag cctccaagaa ggctgatgcc
     7081 gttttgttag gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag acctgaacaa
     7141 ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc atgtaacttt
     7201 gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa aggtactgac
     7261 ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa ggaagacgat
     7321 ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca aagaatcaca
     7381 agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg gtccttggat
     7441 aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga aaccatcaag
     7501 aacgaattcc ctacattgaa ggttcaacat caattgattg attctgccgc catgatccta
     7561 gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt tggtgatatc
     7621 atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc tgcgtccttg
     7681 gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca cggttctgct
     7741 ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg
     7801 ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag
     7861 gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac caccgaagtc
     7921 ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aaaaagattc tcttttttta
     7981 tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac gaaattacaa
     8041 aatggaatat gttcataggg tagacgaaac tatatacgca atctacatac atttatcaag
     8101 aaggagaaaa aggaggatgt aaaggaatac aggtaagcaa attgatacta atggctcaac
     8161 gtgataagga aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac
     8221 aaaaagttag gtgtaacaga aaatcatgaa actatgattc ctaatttata tattggagga
     8281 ttttctctaa aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct gaccatttga
     8341 tggagtttaa gtcaatacct tcttgaacca tttcccataa tggtgaaagt tccctcaaga
     8401 attttactct gtcagaaacg gccttaacga cgtagtcgat atggtgcact ctcagtacaa
     8461 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     8521 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgaca aagggcctcg
     8581 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag gacggatcgc
     8641 ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact
     8701 ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag
     8761 aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
     8821 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
     8881 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
     8941 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
     9001 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     9061 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     9121 atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga
     9181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     9241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     9301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     9361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     9421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     9481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     9541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     9601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     9661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     9721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     9781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     9841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     9901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     9961 tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat tgcagcactg
    10021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    10081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
    10141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
    10201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
    10261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    10321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    10381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    10441 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    10501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
    10561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
    10621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    10681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
    10741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    10801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    10861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
    10921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
    10981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
    11041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
    11101 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
    11161 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
    11221 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
    11281 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
    11341 gaacaaaagc tggagct
//
