LOCUS       sequence-102116-       17106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9, MCP-VP64, and PCP-VP64 in Yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17106)
  AUTHORS   Zalatan JG, Lee ME, Almeida R, Gilbert LA, Whitehead EH, La Russa M,
            Tsai JC, Weissman JS, Dueber JE, Qi LS, Lim WA
  TITLE     Engineering Complex Synthetic Transcriptional Programs with CRISPR 
            RNA Scaffolds.
  JOURNAL   Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 
            10.1016/j.cell.2014.11.052.
  PUBMED    25533786
REFERENCE   2  (bases 1 to 17106)
  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: "Cell. 2014 
            Dec 18. pii: S0092-8674(14)01570-0. doi: 
            10.1016/j.cell.2014.11.052."
COMMENT     Sequence Label: pJZC620
FEATURES             Location/Qualifiers
     source          1..17106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        3292..3996
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4024..4044
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4417..4566
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     promoter        5777..6481
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             6509..6529
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6917..7066
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     terminator      7076..7375
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     promoter        7494..8160
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     8030..8053
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             join(8170..8193,8194..8196,8197..10683,10684..10686,
                     10687..12270)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             12283..12303
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12307..12327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12331..12351
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(13653..13672)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(13654..13672)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13679..13696)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13679..13695)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13688..13710)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      13836..14291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13923..13942)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14133..14154
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(14647..14666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14766..14788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(14826..14844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        14929..15033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(15034..15102,15103..15894)
                     /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(15252..15271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      16065..16653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     16554..16573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     16807..16824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    16941..16962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(16977..16982,16983..17000,17001..17007)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    17015..17031
                     /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     17020..17042
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     17039..17055
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     17039..17055
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        17076..17094
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc aaaatccaca
      121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
      181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
      241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
      301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
      361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
      421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
      481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
      541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
      601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
      661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa
      721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa
      781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc
      841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg
      901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa
      961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt
     1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct
     1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc
     1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat
     1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga
     1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa
     1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga
     1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc
     1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt
     1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc
     1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg
     1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga
     1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca
     1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag
     1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca
     1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg
     1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc
     1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc
     2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa
     2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga
     2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc
     2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag
     2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga
     2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc
     2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat
     2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggcccta aaacaagaag
     2521 agggttgact acatcacgat gagggggatc gaagaaatga tggtaaatga aataggaaat
     2581 caaggagcat gaaggcaaaa gacaaatata agggtcgaac gaaaaataaa gtgaaaagtg
     2641 ttgatatgat gtatttggct ttgcggcgcc gaaaaaacga gtttacgcaa ttgcacaatc
     2701 atgctgactc tgtggcggac ccgcgctctt gccggcccgg cgataacgct gggcgtgagg
     2761 ctgtgcccgg cggagttttt tgcgcctgca ttttccaagg tttaccctgc gctaaggggc
     2821 gagattggag aagcaataag aatgccggtt ggggttgcga tgatgacgac cacgacaact
     2881 ggtgtcatta tttaagttgc cgaaagaacc tgagtgcatt tgcaacatga gtatactaga
     2941 agaatgagcc aagacttgcg agacgcgagt ttgccggtgg tgcgaacaat agagcgacca
     3001 tgaccttgaa ggtgagacgc gcataaccgc tagagtactt tgaagaggaa acagcaatag
     3061 ggttgctacc agtataaata gacaggtaca tacaacactg gaaatggttg tctgtttgag
     3121 tacgctttca attcatttgg gtgtgcactt tattatgtta caatatggaa gggaacttta
     3181 cacttctcct atgcacatat attaattaaa gtccaatgct agtagagaag gggggtaaca
     3241 cccctccgcg ctcttttccg atttttttct aaaccgtgga atatttcgga tatccttttg
     3301 ttgtttccgg gtgtacaata tggacttcct cttttctggc aaccaaaccc atacatcggg
     3361 attcctataa taccttcgtt ggtctcccta acatgtaggt ggcggagggg agatatacaa
     3421 tagaacagat accagacagg acataatggg ctaaacaaga ctacaccaat tacactgcct
     3481 cattgatggt ggtacataac gaactaatac tgtagcccta gacttgatag ccatcatcat
     3541 atcgaagttt cactaccctt tttccatttg ccatctattg aagtaataat aggcgcatgc
     3601 aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa
     3661 tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca
     3721 gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc
     3781 ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt
     3841 gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta
     3901 ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc
     3961 acaatatttc aagctatacc aagcatacaa tcaactatct catatacaat gtctctcgag
     4021 atgcccaaaa agaaaagaaa agtgggtagt atggcttcta actttactca gttcgttctc
     4081 gtcgacaatg gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacgggatc
     4141 gctgaatgga tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt
     4201 cagagctctg cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa aggcgcctgg
     4261 cgttcgtact taaatatgga actaaccatt ccaattttcg ccacgaattc cgactgcgag
     4321 cttattgtta aggcaatgca aggtctccta aaagatggaa acccgattcc ctcagcaatc
     4381 gcagcaaact ccggcatcta cggctccgga cgagccgatg ctctcgacga ttttgacctc
     4441 gatatgctgg gatccgacgc tctcgatgac ttcgaccttg acatgctggg gagtgacgct
     4501 ctggacgact tcgatctcga catgctgggc tccgacgccc tggatgactt cgatctggac
     4561 atgctcatca actgaagtgc ttttaactaa gaattattag tcttttctgc ttattttttc
     4621 atcatagttt agaacacttt atattaacga atagtttatg aatctattta ggtttaaaaa
     4681 ttgatacagt tttataagtt actttttcaa agactcgtgc tgtctattgc ataatgcact
     4741 ggaaggggaa aaaaaaggtg cacacgcgtg gctttttctt gaatttgcag tttgaaaaat
     4801 aactacatgg atgataagaa aacatggagt acagtcactt tgagaacctt caatcagctg
     4861 gtaacgtctt cgttaattgg atactcaaaa aagatggata gcatgaatca caagatggaa
     4921 ggaaatgcgg gccacgacca cagtgatatg catatgggag atggagatga tacctgacta
     4981 ctgcacgcta cgctaaaaca agaagagggt tgactacatc acgatgaggg ggatcgaaga
     5041 aatgatggta aatgaaatag gaaatcaagg agcatgaagg caaaagacaa atataagggt
     5101 cgaacgaaaa ataaagtgaa aagtgttgat atgatgtatt tggctttgcg gcgccgaaaa
     5161 aacgagttta cgcaattgca caatcatgct gactctgtgg cggacccgcg ctcttgccgg
     5221 cccggcgata acgctgggcg tgaggctgtg cccggcggag ttttttgcgc ctgcattttc
     5281 caaggtttac cctgcgctaa ggggcgagat tggagaagca ataagaatgc cggttggggt
     5341 tgcgatgatg acgaccacga caactggtgt cattatttaa gttgccgaaa gaacctgagt
     5401 gcatttgcaa catgagtata ctagaagaat gagccaagac ttgcgagacg cgagtttgcc
     5461 ggtggtgcga acaatagagc gaccatgacc ttgaaggtga gacgcgcata accgctagag
     5521 tactttgaag aggaaacagc aatagggttg ctaccagtat aaatagacag gtacatacaa
     5581 cactggaaat ggttgtctgt ttgagtacgc tttcaattca tttgggtgtg cactttatta
     5641 tgttacaata tggaagggaa ctttacactt ctcctatgca catatattaa ttaaagtcca
     5701 atgctagtag agaagggggg taacacccct ccgcgctctt ttccgatttt tttctaaacc
     5761 gtggaatatt tcggatatcc ttttgttgtt tccgggtgta caatatggac ttcctctttt
     5821 ctggcaacca aacccataca tcgggattcc tataatacct tcgttggtct ccctaacatg
     5881 taggtggcgg aggggagata tacaatagaa cagataccag acaggacata atgggctaaa
     5941 caagactaca ccaattacac tgcctcattg atggtggtac ataacgaact aatactgtag
     6001 ccctagactt gatagccatc atcatatcga agtttcacta ccctttttcc atttgccatc
     6061 tattgaagta ataataggcg catgcaactt cttttctttt tttttctttt ctctctcccc
     6121 cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa
     6181 attaacgaca aagacagcac caacagatgt cgttgttcca gagctgatga ggggtatctc
     6241 gaagcacacg aaactttttc cttccttcat tcacgcacac tactctctaa tgagcaacgg
     6301 tatacggcct tccttccagt tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta
     6361 tcaagtataa atagacctgc aattattaat cttttgtttc ctcgtcattg ttctcgttcc
     6421 ctttcttcct tgtttctttt tctgcacaat atttcaagct ataccaagca tacaatcaac
     6481 tatctcatat acaatgtctc tcgagatgcc caaaaagaaa agaaaagtgg gtagtatgtc
     6541 caaaaccatc gttctttcgg tcggcgaggc tactcgcact ctgactgaga tccagtccac
     6601 cgcagaccgt cagatcttcg aagagaaggt cgggcctctg gtgggtcggc tgcgcctcac
     6661 ggcttcgctc cgtcaaaacg gagccaagac cgcgtatcgc gtcaacctaa aactggatca
     6721 ggcggacgtc gttgattccg gacttccgaa agtgcgctac actcaggtat ggtcgcacga
     6781 cgtgacaatc gttgcgaata gcaccgaggc ctcgcgcaaa tcgttgtacg atttgaccaa
     6841 gtccctcgtc gcgacctcgc aggtcgaaga tcttgtcgtc aaccttgtgc cgctgggccg
     6901 tggctccgga cgagccgatg ctctcgacga ttttgacctc gatatgctgg gatccgacgc
     6961 tctcgatgac ttcgaccttg acatgctggg gagtgacgct ctggacgact tcgatctcga
     7021 catgctgggc tccgacgccc tggatgactt cgatctggac atgctcatca actgagcgga
     7081 tctcttatgt ctttacgatt tatagttttc attatcaagt atgcctatat tagtatatag
     7141 catctttaga tgacagtgtt cgaagtttca cgaataaaag ataatattct actttttgct
     7201 cccaccgcgt ttgctagcac gagtgaacac catccctcgc ctgtgagttg tacccattcc
     7261 tctaaactgt agacatggta gcttcagcag tgttcgttat gtacggcatc ctccaacaaa
     7321 cagtcggtta tagtttgtcc tgctcctctg aatcgtctcc ctcgatattt ctcattttcc
     7381 ttcgcatgcc agcattgaaa tgatcgaagt tcaatgatga aacggtaatt cttctgtcat
     7441 ttactcatct catctcatca agttatataa ttctaggccc cagttcgagt ttatcattat
     7501 caatactgcc atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat
     7561 ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc
     7621 gggttacaca gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc
     7681 actaaatata atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca
     7741 ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta
     7801 cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc
     7861 tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct
     7921 tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa
     7981 accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt
     8041 gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct
     8101 tttttttagt tttaaaacac caagaactta gtttcgaata aacacacata aacaaacaaa
     8161 atgctcgagg acaagaagta ttctatcgga ctggccatcg ggactaatag cgtcgggtgg
     8221 gccgtgatca ctgacgagta caaggtgccc tctaagaagt tcaaggtgct cgggaacacc
     8281 gaccggcatt ccatcaagaa aaatctgatc ggagctctcc tctttgattc aggggagacc
     8341 gctgaagcaa cccgcctcaa gcggactgct agacggcggt acaccaggag gaagaaccgg
     8401 atttgttacc ttcaagagat attctccaac gaaatggcaa aggtcgacga cagcttcttc
     8461 cataggctgg aagaatcatt cctcgtggaa gaggataaga agcatgaacg gcatcccatc
     8521 ttcggtaata tcgtcgacga ggtggcctat cacgagaaat acccaaccat ctaccatctt
     8581 cgcaaaaagc tggtggactc aaccgacaag gcagacctcc ggcttatcta cctggccctg
     8641 gcccacatga tcaagttcag aggccacttc ctgatcgagg gcgacctcaa tcctgacaat
     8701 agcgatgtgg ataaactgtt catccagctg gtgcagactt acaaccagct ctttgaagag
     8761 aaccccatca atgcaagcgg agtcgatgcc aaggccattc tgtcagcccg gctgtcaaag
     8821 agccgcagac ttgagaatct tatcgctcag ctgccgggtg aaaagaaaaa tggactgttc
     8881 gggaacctga ttgctctttc acttgggctg actcccaatt tcaagtctaa tttcgacctg
     8941 gcagaggatg ccaagctgca actgtccaag gacacctatg atgacgatct cgacaacctc
     9001 ctggcccaga tcggtgacca atacgccgac cttttccttg ctgctaagaa tctttctgac
     9061 gccatcctgc tgtctgacat tctccgcgtg aacactgaaa tcaccaaggc ccctctttca
     9121 gcttcaatga ttaagcggta tgatgagcac caccaggacc tgaccctgct taaggcactc
     9181 gtccggcagc agcttccgga gaagtacaag gaaatcttct ttgaccagtc aaagaatgga
     9241 tacgccggct acatcgacgg aggtgcctcc caagaggaat tttataagtt tatcaaacct
     9301 atccttgaga agatggacgg caccgaagag ctcctcgtga aactgaatcg ggaggatctg
     9361 ctgcggaagc agcgcacttt cgacaatggg agcattcccc accagatcca tcttggggag
     9421 cttcacgcca tccttcggcg ccaagaggac ttctacccct ttcttaagga caacagggag
     9481 aagattgaga aaattctcac tttccgcatc ccctactacg tgggacccct cgccagagga
     9541 aatagccggt ttgcttggat gaccagaaag tcagaagaaa ctatcactcc ctggaacttc
     9601 gaagaggtgg tggacaaggg agccagcgct cagtcattca tcgaacggat gactaacttc
     9661 gataagaacc tccccaatga gaaggtcctg ccgaaacatt ccctgctcta cgagtacttt
     9721 accgtgtaca acgagctgac caaggtgaaa tatgtcaccg aagggatgag gaagcccgca
     9781 ttcctgtcag gcgaacaaaa gaaggcaatt gtggaccttc tgttcaagac caatagaaag
     9841 gtgaccgtga agcagctgaa ggaggactat ttcaagaaaa ttgaatgctt cgactctgtg
     9901 gagattagcg gggtcgaaga tcggttcaac gcaagcctgg gtacctacca tgatctgctt
     9961 aagatcatca aggacaagga ttttctggac aatgaggaga acgaggacat ccttgaggac
    10021 attgtcctga ctctcactct gttcgaggac cgggaaatga tcgaggagag gcttaagacc
    10081 tacgcccatc tgttcgacga taaagtgatg aagcaactta aacggagaag atataccgga
    10141 tggggacgcc ttagccgcaa actcatcaac ggaatccggg acaaacagag cggaaagacc
    10201 attcttgatt tccttaagag cgacggattc gctaatcgca acttcatgca acttatccat
    10261 gatgattccc tgacctttaa ggaggacatc cagaaggccc aagtgtctgg acaaggtgac
    10321 tcactgcacg agcatatcgc aaatctggct ggttcacccg ctattaagaa gggtattctc
    10381 cagaccgtga aagtcgtgga cgagctggtc aaggtgatgg gtcgccataa accagagaac
    10441 attgtcatcg agatggccag ggaaaaccag actacccaga agggacagaa gaacagcagg
    10501 gagcggatga aaagaattga ggaagggatt aaggagctcg ggtcacagat ccttaaagag
    10561 cacccggtgg aaaacaccca gcttcagaat gagaagctct atctgtacta ccttcaaaat
    10621 ggacgcgata tgtatgtgga ccaagagctt gatatcaaca ggctctcaga ctacgacgtg
    10681 gacgccatcg tccctcagag cttcctcaaa gacgactcaa ttgacaataa ggtgctgact
    10741 cgctcagaca agaaccgggg aaagtcagat aacgtgccct cagaggaagt cgtgaaaaag
    10801 atgaagaact attggcgcca gcttctgaac gcaaagctga tcactcagcg gaagttcgac
    10861 aatctcacta aggctgagag gggcggactg agcgaactgg acaaagcagg attcattaaa
    10921 cggcaacttg tggagactcg gcagattact aaacatgtcg cccaaatcct tgactcacgc
    10981 atgaatacca agtacgacga aaacgacaaa cttatccgcg aggtgaaggt gattaccctg
    11041 aagtccaagc tggtcagcga tttcagaaag gactttcaat tctacaaagt gcgggagatc
    11101 aataactatc atcatgctca tgacgcatat ctgaatgccg tggtgggaac cgccctgatc
    11161 aagaagtacc caaagctgga aagcgagttc gtgtacggag actacaaggt ctacgacgtg
    11221 cgcaagatga ttgccaaatc tgagcaggag atcggaaagg ccaccgcaaa gtacttcttc
    11281 tacagcaaca tcatgaattt cttcaagacc gaaatcaccc ttgcaaacgg tgagatccgg
    11341 aagaggccgc tcatcgagac taatggggag actggcgaaa tcgtgtggga caagggcaga
    11401 gatttcgcta ccgtgcgcaa agtgctttct atgcctcaag tgaacatcgt gaagaaaacc
    11461 gaggtgcaaa ccggaggctt ttctaaggaa tcaatcctcc ccaagcgcaa ctccgacaag
    11521 ctcattgcaa ggaagaagga ttgggaccct aagaagtacg gcggattcga ttcaccaact
    11581 gtggcttatt ctgtcctggt cgtggctaag gtggaaaaag gaaagtctaa gaagctcaag
    11641 agcgtgaagg aactgctggg tatcaccatt atggagcgca gctccttcga gaagaaccca
    11701 attgactttc tcgaagccaa aggttacaag gaagtcaaga aggaccttat catcaagctc
    11761 ccaaagtata gcctgttcga actggagaat gggcggaagc ggatgctcgc ctccgctggc
    11821 gaacttcaga agggtaatga gctggctctc ccctccaagt acgtgaattt cctctacctt
    11881 gcaagccatt acgagaagct gaaggggagc cccgaggaca acgagcaaaa gcaactgttt
    11941 gtggagcagc ataagcatta tctggacgag atcattgagc agatttccga gttttctaaa
    12001 cgcgtcattc tcgctgatgc caacctcgat aaagtcctta gcgcatacaa taagcacaga
    12061 gacaaaccaa ttcgggagca ggctgagaat atcatccacc tgttcaccct caccaatctt
    12121 ggtgcccctg ccgcattcaa gtacttcgac accaccatcg accggaaacg ctatacctcc
    12181 accaaagaag tgctggacgc caccctcatc caccagagca tcaccggact ttacgaaact
    12241 cggattgacc tctcacagct cggaggggat gagggagctg atccaaaaaa gaagagaaag
    12301 gtagatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt atagaattct
    12361 accggcggcc gccaccgcgg tggagctcta agcaaatagc taaattatat acgaattaat
    12421 attatgatta agtgtttacg tgagtgcgat atttttatta ctatcttata cagttgtata
    12481 tactctataa aatgagttgt ctattaatta acgcgatgaa tgctttctgg gtttacctct
    12541 ccaacaactc tagtttactt ctcaatacat tcaattgtat ttgatttgtc aatacttcat
    12601 cattaatcaa ttctatagtt ttgtttttct cgtttatttc caaatttaat gcatcaattt
    12661 tattattcaa tttgtcgttg attttggtta atgattttat ggtttgatct ctggcattga
    12721 ttgtttgtgt tagtttttca ttattgataa ttaaattatt taagttagtt atcaactcgg
    12781 tgttttcaag tttcaagttt tcaatttctt tagagtttat tagatttgtc aaagtttctg
    12841 aattgcttga ttggtcctgt agaagagtat ttgttgttgt ggataattga ttcaattttt
    12901 gagacaattg ctggaaggcg ttgaaatatc tagcatcaat ctcatggttt ttttcccgag
    12961 agtctcgtag attcaattgt tttaatatat cttgggacca ctcttgattt gaactcatgg
    13021 aaattaaact gggtgttgtg ttgtggtgta atgattgtac cccctttgct tataattgtg
    13081 tggccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag
    13141 attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg
    13201 acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac
    13261 atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat
    13321 actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga
    13381 ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt
    13441 tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg
    13501 acctgaccat ttgatggagt ttaagtcaat accttcttga accatttccc ataatggtga
    13561 aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtagg ccggccattt
    13621 aaatgtttaa acggccacgc tggcccaatt cgccctatag tgagtcgtat tacaattcac
    13681 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
    13741 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
    13801 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
    13861 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
    13921 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
    13981 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
    14041 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
    14101 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
    14161 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
    14221 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
    14281 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
    14341 atagggtaat aactgatata attaaattga agctctaatt tgtgagttta gtattaggat
    14401 ctctaccctt ggcgaaaagt cctctgccaa caatgatgat atctgatcca ccacttacaa
    14461 cttcgtcgac ggttctgtac tgctgaccca atatgcgtat atataccaat ctaagtctgt
    14521 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
    14581 accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac
    14641 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
    14701 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
    14761 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
    14821 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
    14881 gacgtcggcg cgccgaggac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
    14941 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
    15001 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
    15061 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
    15121 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
    15181 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
    15241 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
    15301 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
    15361 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
    15421 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    15481 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    15541 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    15601 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    15661 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    15721 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    15781 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    15841 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    15901 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    15961 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    16021 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    16081 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    16141 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    16201 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    16261 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    16321 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    16381 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    16441 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    16501 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    16561 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    16621 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    16681 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    16741 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    16801 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    16861 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    16921 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    16981 cactttatgc ttccggctcc tatgttgtgt ggaattgtga gcggataaca atttcacaca
    17041 ggaaacagct atgaccatga ttacgccaag ctcggaatta accctcacta aagggaacaa
    17101 aagctg
//
