LOCUS       sequence-484247-       15193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast vector for integration of pGAL10-dCas9 and pGAL10-MCP-VPR at 
            LEU2 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15193)
  AUTHORS   Stohr AM, Hansen H, Richards B, Park H, Goncalves AG, Agrawal A, 
            Blenner M, Chen W
  TITLE     Metabolite-responsive scaffold RNAs for dynamic CRISPR 
            transcriptional regulation.
  JOURNAL   Nucleic Acids Res. 2025 Nov 13;53(21):gkaf1290. doi: 
            10.1093/nar/gkaf1290.
  PUBMED    41277686
REFERENCE   2  (bases 1 to 15193)
  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: "Nucleic 
            Acids Res."; date: "2025-11-13"; volume: "53(21):gkaf1290. doi"; 
            pages: " 10.1093/nar/gkaf1290"
COMMENT     Sequence Label: pSC-pGAL10-CRISPRa
FEATURES             Location/Qualifiers
     source          1..15193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        2509..3173
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2840..2957
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             3200..3220
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3593..3742
                     /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"
     CDS             3767..3787
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4217..4573
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             4592..5161
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     promoter        5580..6244
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    5911..6028
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             join(6265..6288,6289..6291,6292..8778,8779..8781,
                     8782..10365)
                     /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             10378..10398
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10402..10422
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10426..10446
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(11740..11759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11741..11759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11766..11783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11766..11782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11775..11797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11923..12378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12010..12029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12220..12241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12734..12753)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12853..12875
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12913..12931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13016..13120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13121..13189,13190..13981)
                     /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(13339..13358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14152..14740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14641..14660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14894..14911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    15028..15049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(15064..15069,15070..15087,15088..15094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15102..15118
                     /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     15107..15129
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     15126..15142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15126..15142
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        15163..15181
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa cgattacatc 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 ccgggcccgt tttttctcct
     2521 tgacgttaaa gtatagaggt atattaacaa ttttttgttg atacttttat tacatttgaa
     2581 taagaagtaa tacaaaccga aaatgttgaa agtattagtt aaagtggtta tgcagttttt
     2641 gcatttatat atctgttaat agatcaaaaa tcatcgcttc gctgattaat taccccagaa
     2701 ataaggctaa aaaactaatc gcattatcat cctatggttg ttaatttgat tcgttcattt
     2761 gaaggtttgt ggggccaggt tactgccaat ttttcctctt cataaccata aaagctagta
     2821 ttgtagaatc tttattgttc ggagcagtgc ggcgcgaggc acatctgcgt ttcaggaacg
     2881 cgaccggtga agacgaggac gcacggagga gagtcttcct tcggagggct gtcacccgct
     2941 cggcggcttc taatccgtac ttcaatatag caatgagcag ttaagcgtat tactgaaagt
     3001 tccaaagaga aggttttttt aggctaagat aatggggctc tttacatttc cacaacatat
     3061 aagtaagatt agatatggat atgtatatgg atatgtatat ggtggtaatg ccatgtaata
     3121 tgattattaa acttctttgc gtccatccaa aaaaaaagta agaatttttg aaaattcaat
     3181 ataaatgtct ctcgagatgc ccaaaaagaa aagaaaagtg ggtagtatgg cttctaactt
     3241 tactcagttc gttctcgtcg acaatggcgg aactggcgac gtgactgtcg ccccaagcaa
     3301 cttcgctaac gggatcgctg aatggatcag ctctaactcg cgttcacagg cttacaaagt
     3361 aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa tacaccatca aagtcgaggt
     3421 gcctaaaggc gcctggcgtt cgtacttaaa tatggaacta accattccaa ttttcgccac
     3481 gaattccgac tgcgagctta ttgttaaggc aatgcaaggt ctcctaaaag atggaaaccc
     3541 gattccctca gcaatcgcag caaactccgg catctacggc tccggacgag ccgatgctct
     3601 cgacgatttt gacctcgata tgctgggatc cgacgctctc gatgacttcg accttgacat
     3661 gctggggagt gacgctctgg acgacttcga tctcgacatg ctgggctccg acgccctgga
     3721 tgacttcgat ctggacatgc tcatcaactc tagaagttcc ggatctccga aaaagaaacg
     3781 caaagttggt agccagtacc tgcccgacac cgacgaccgg caccggatcg aggaaaagcg
     3841 gaagcggacc tacgagacat tcaagagcat catgaagaag tcccccttca gcggccccac
     3901 cgaccctaga cctccaccta gaagaatcgc cgtgcccagc agatccagcg ccagcgtgcc
     3961 aaaacctgcc ccccagcctt accccttcac cagcagcctg agcaccatca actacgacga
     4021 gttccctacc atggtgttcc ccagcggcca gatctctcag gcctctgctc tggctccagc
     4081 ccctcctcag gtgctgcctc aggctcctgc tcctgcacca gctccagcca tggtgtctgc
     4141 actggctcag gcaccagcac ccgtgcctgt gctggctcct ggacctccac aggctgtggc
     4201 tccaccagcc cctaaaccta cacaggccgg cgagggcaca ctgtctgaag ctctgctgca
     4261 gctgcagttc gacgacgagg atctgggagc cctgctggga aacagcaccg atcctgccgt
     4321 gttcaccgac ctggccagcg tggacaacag cgagttccag cagctgctga accagggcat
     4381 ccctgtggcc cctcacacca ccgagcccat gctgatggaa taccccgagg ccatcacccg
     4441 gctcgtgaca ggcgctcaga ggcctcctga tccagctcct gcccctctgg gagcaccagg
     4501 cctgcctaat ggactgctgt ctggcgacga ggacttcagc tctatcgccg acatggactt
     4561 ctccgcactg ctgggtagcg gatcgggatc tcgggattcc agggaaggga tgtttttgcc
     4621 gaagcctgag gccggctccg ctattagtga cgtgtttgag ggccgcgagg tgtgccagcc
     4681 aaaacgaatc cggccatttc atcctccagg aagtccatgg gccaaccgcc cactccccgc
     4741 cagcctcgca ccaacaccaa ccggtccagt acatgagcca gtcgggtcac tgaccccggc
     4801 accagtccct cagccactgg atccagcgcc cgcagtgact cccgaggcca gtcacctgtt
     4861 ggaggatccc gatgaagaga cgagccaggc tgtcaaagcc cttcgggaga tggccgatac
     4921 tgtgattccc cagaaggaag aggctgcaat ctgtggccaa atggaccttt cccatccgcc
     4981 cccaaggggc catctggatg agctgacaac cacacttgag tccatgaccg aggatctgaa
     5041 cctggactca cccctgaccc cggaattgaa cgagattctg gataccttcc tgaacgacga
     5101 gtgcctcttg catgccatgc atatcagcac aggactgtcc atcttcgaca catctctgtt
     5161 ttgaagtgct tttaactaag aattattagt cttttctgct tattttttca tcatagttta
     5221 gaacacttta tattaacgaa tagtttatga atctatttag gtttaaaaat tgatacagtt
     5281 ttataagtta ctttttcaaa gactcgtgct gtctattgca taatgcactg gaaggggaaa
     5341 aaaaaggtgc acacgcgtgg ctttttcttg aatttgcagt ttgaaaaata actacatgga
     5401 tgataagaaa acatggagta cagtcacttt gagaaccttc aatcagctgg taacgtcttc
     5461 gttaattgga tactcaaaaa agatggatag catgaatcac aagatggaag gaaatgcggg
     5521 ccacgaccac agtgatatgc atatgggaga tggagatgat acctggcccc ggtgctatag
     5581 ttttttctcc ttgacgttaa agtatagagg tatattaaca attttttgtt gatactttta
     5641 ttacatttga ataagaagta atacaaaccg aaaatgttga aagtattagt taaagtggtt
     5701 atgcagtttt tgcatttata tatctgttaa tagatcaaaa atcatcgctt cgctgattaa
     5761 ttaccccaga aataaggcta aaaaactaat cgcattatca tcctatggtt gttaatttga
     5821 ttcgttcatt tgaaggtttg tggggccagg ttactgccaa tttttcctct tcataaccat
     5881 aaaagctagt attgtagaat ctttattgtt cggagcagtg cggcgcgagg cacatctgcg
     5941 tttcaggaac gcgaccggtg aagacgagga cgcacggagg agagtcttcc ttcggagggc
     6001 tgtcacccgc tcggcggctt ctaatccgta cttcaatata gcaatgagca gttaagcgta
     6061 ttactgaaag ttccaaagag aaggtttttt taggctaaga taatggggct ctttacattt
     6121 ccacaacata taagtaagat tagatatgga tatgtatatg gatatgtata tggtggtaat
     6181 gccatgtaat atgattatta aacttctttg cgtccatcca aaaaaaaagt aagaattttt
     6241 gaaaattcaa tataaatgct cgaggacaag aagtattcta tcggactggc catcgggact
     6301 aatagcgtcg ggtgggccgt gatcactgac gagtacaagg tgccctctaa gaagttcaag
     6361 gtgctcggga acaccgaccg gcattccatc aagaaaaatc tgatcggagc tctcctcttt
     6421 gattcagggg agaccgctga agcaacccgc ctcaagcgga ctgctagacg gcggtacacc
     6481 aggaggaaga accggatttg ttaccttcaa gagatattct ccaacgaaat ggcaaaggtc
     6541 gacgacagct tcttccatag gctggaagaa tcattcctcg tggaagagga taagaagcat
     6601 gaacggcatc ccatcttcgg taatatcgtc gacgaggtgg cctatcacga gaaataccca
     6661 accatctacc atcttcgcaa aaagctggtg gactcaaccg acaaggcaga cctccggctt
     6721 atctacctgg ccctggccca catgatcaag ttcagaggcc acttcctgat cgagggcgac
     6781 ctcaatcctg acaatagcga tgtggataaa ctgttcatcc agctggtgca gacttacaac
     6841 cagctctttg aagagaaccc catcaatgca agcggagtcg atgccaaggc cattctgtca
     6901 gcccggctgt caaagagccg cagacttgag aatcttatcg ctcagctgcc gggtgaaaag
     6961 aaaaatggac tgttcgggaa cctgattgct ctttcacttg ggctgactcc caatttcaag
     7021 tctaatttcg acctggcaga ggatgccaag ctgcaactgt ccaaggacac ctatgatgac
     7081 gatctcgaca acctcctggc ccagatcggt gaccaatacg ccgacctttt ccttgctgct
     7141 aagaatcttt ctgacgccat cctgctgtct gacattctcc gcgtgaacac tgaaatcacc
     7201 aaggcccctc tttcagcttc aatgattaag cggtatgatg agcaccacca ggacctgacc
     7261 ctgcttaagg cactcgtccg gcagcagctt ccggagaagt acaaggaaat cttctttgac
     7321 cagtcaaaga atggatacgc cggctacatc gacggaggtg cctcccaaga ggaattttat
     7381 aagtttatca aacctatcct tgagaagatg gacggcaccg aagagctcct cgtgaaactg
     7441 aatcgggagg atctgctgcg gaagcagcgc actttcgaca atgggagcat tccccaccag
     7501 atccatcttg gggagcttca cgccatcctt cggcgccaag aggacttcta cccctttctt
     7561 aaggacaaca gggagaagat tgagaaaatt ctcactttcc gcatccccta ctacgtggga
     7621 cccctcgcca gaggaaatag ccggtttgct tggatgacca gaaagtcaga agaaactatc
     7681 actccctgga acttcgaaga ggtggtggac aagggagcca gcgctcagtc attcatcgaa
     7741 cggatgacta acttcgataa gaacctcccc aatgagaagg tcctgccgaa acattccctg
     7801 ctctacgagt actttaccgt gtacaacgag ctgaccaagg tgaaatatgt caccgaaggg
     7861 atgaggaagc ccgcattcct gtcaggcgaa caaaagaagg caattgtgga ccttctgttc
     7921 aagaccaata gaaaggtgac cgtgaagcag ctgaaggagg actatttcaa gaaaattgaa
     7981 tgcttcgact ctgtggagat tagcggggtc gaagatcggt tcaacgcaag cctgggtacc
     8041 taccatgatc tgcttaagat catcaaggac aaggattttc tggacaatga ggagaacgag
     8101 gacatccttg aggacattgt cctgactctc actctgttcg aggaccggga aatgatcgag
     8161 gagaggctta agacctacgc ccatctgttc gacgataaag tgatgaagca acttaaacgg
     8221 agaagatata ccggatgggg acgccttagc cgcaaactca tcaacggaat ccgggacaaa
     8281 cagagcggaa agaccattct tgatttcctt aagagcgacg gattcgctaa tcgcaacttc
     8341 atgcaactta tccatgatga ttccctgacc tttaaggagg acatccagaa ggcccaagtg
     8401 tctggacaag gtgactcact gcacgagcat atcgcaaatc tggctggttc acccgctatt
     8461 aagaagggta ttctccagac cgtgaaagtc gtggacgagc tggtcaaggt gatgggtcgc
     8521 cataaaccag agaacattgt catcgagatg gccagggaaa accagactac ccagaaggga
     8581 cagaagaaca gcagggagcg gatgaaaaga attgaggaag ggattaagga gctcgggtca
     8641 cagatcctta aagagcaccc ggtggaaaac acccagcttc agaatgagaa gctctatctg
     8701 tactaccttc aaaatggacg cgatatgtat gtggaccaag agcttgatat caacaggctc
     8761 tcagactacg acgtggacgc catcgtccct cagagcttcc tcaaagacga ctcaattgac
     8821 aataaggtgc tgactcgctc agacaagaac cggggaaagt cagataacgt gccctcagag
     8881 gaagtcgtga aaaagatgaa gaactattgg cgccagcttc tgaacgcaaa gctgatcact
     8941 cagcggaagt tcgacaatct cactaaggct gagaggggcg gactgagcga actggacaaa
     9001 gcaggattca ttaaacggca acttgtggag actcggcaga ttactaaaca tgtcgcccaa
     9061 atccttgact cacgcatgaa taccaagtac gacgaaaacg acaaacttat ccgcgaggtg
     9121 aaggtgatta ccctgaagtc caagctggtc agcgatttca gaaaggactt tcaattctac
     9181 aaagtgcggg agatcaataa ctatcatcat gctcatgacg catatctgaa tgccgtggtg
     9241 ggaaccgccc tgatcaagaa gtacccaaag ctggaaagcg agttcgtgta cggagactac
     9301 aaggtctacg acgtgcgcaa gatgattgcc aaatctgagc aggagatcgg aaaggccacc
     9361 gcaaagtact tcttctacag caacatcatg aatttcttca agaccgaaat cacccttgca
     9421 aacggtgaga tccggaagag gccgctcatc gagactaatg gggagactgg cgaaatcgtg
     9481 tgggacaagg gcagagattt cgctaccgtg cgcaaagtgc tttctatgcc tcaagtgaac
     9541 atcgtgaaga aaaccgaggt gcaaaccgga ggcttttcta aggaatcaat cctccccaag
     9601 cgcaactccg acaagctcat tgcaaggaag aaggattggg accctaagaa gtacggcgga
     9661 ttcgattcac caactgtggc ttattctgtc ctggtcgtgg ctaaggtgga aaaaggaaag
     9721 tctaagaagc tcaagagcgt gaaggaactg ctgggtatca ccattatgga gcgcagctcc
     9781 ttcgagaaga acccaattga ctttctcgaa gccaaaggtt acaaggaagt caagaaggac
     9841 cttatcatca agctcccaaa gtatagcctg ttcgaactgg agaatgggcg gaagcggatg
     9901 ctcgcctccg ctggcgaact tcagaagggt aatgagctgg ctctcccctc caagtacgtg
     9961 aatttcctct accttgcaag ccattacgag aagctgaagg ggagccccga ggacaacgag
    10021 caaaagcaac tgtttgtgga gcagcataag cattatctgg acgagatcat tgagcagatt
    10081 tccgagtttt ctaaacgcgt cattctcgct gatgccaacc tcgataaagt ccttagcgca
    10141 tacaataagc acagagacaa accaattcgg gagcaggctg agaatatcat ccacctgttc
    10201 accctcacca atcttggtgc ccctgccgca ttcaagtact tcgacaccac catcgaccgg
    10261 aaacgctata cctccaccaa agaagtgctg gacgccaccc tcatccacca gagcatcacc
    10321 ggactttacg aaactcggat tgacctctca cagctcggag gggatgaggg agctgatcca
    10381 aaaaagaaga gaaaggtaga tccaaaaaag aagagaaagg tagatccaaa aaagaagaga
    10441 aaggtataga attctaccgg cggccgccac cgcggtggag ctctaagcaa atagctaaat
    10501 tatatacgaa ttaatattat gattaagtgt ttacgtgagt gcgatatttt tattactatc
    10561 ttatacagtt gtatatactc tataaaatga gttgtctatt aattaacgcg atgaatgctt
    10621 tctgggttta cctctccaac aactctagtt tacttctcaa tacattcaat tgtatttgat
    10681 ttgtcaatac ttcatcatta atcaattcta tagttttgtt tttctcgttt atttccaaat
    10741 ttaatgcatc aattttatta ttcaatttgt cgttgatttt ggttaatgat tttatggttt
    10801 gatctctggc attgattgtt tgtgttagtt tttcattatt gataattaaa ttatttaagt
    10861 tagttatcaa ctcggtgttt tcaagtttca agttttcaat ttctttagag tttattagat
    10921 ttgtcaaagt ttctgaattg cttgattggt cctgtagaag agtatttgtt gttgtggata
    10981 attgattcaa tttttgagac aattgctgga aggcgttgaa atatctagca tcaatctcat
    11041 ggtttttttc ccgagagtct cgtagattca attgttttaa tatatcttgg gaccactctt
    11101 gatttgaact catggaaatt aaactgggtg ttgtgttgtg gtgtaatgat tgtaccccct
    11161 ttgcttataa ttgtgtggcc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
    11221 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
    11281 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
    11341 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
    11401 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
    11461 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
    11521 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaat acaacaaata
    11581 aaaaacactc aatgacctga ccatttgatg gagtttaagt caatacctcc ttgaaccatt
    11641 tcccataatg gtgaaagttc cctcaagaat tttactctgt cagaaacggc cttaacgacg
    11701 taggccggcc atttaaatgt ttaaacggcc acgctggccc cctatagtga gtcgtattac
    11761 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
    11821 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
    11881 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
    11941 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
    12001 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
    12061 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
    12121 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
    12181 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
    12241 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
    12301 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
    12361 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
    12421 acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
    12481 ttaggatctc tacccttggc gaaaagtcct ctgccaacaa tgatgatatc tgatccacca
    12541 cttacaactt cgtcgacggt tctgtactgc tgacccaata tgcgtatata taccaatcta
    12601 agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt
    12661 caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat
    12721 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
    12781 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
    12841 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
    12901 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
    12961 tttcttagac gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg
    13021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
    13081 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
    13141 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
    13201 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
    13261 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
    13321 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
    13381 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
    13441 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
    13501 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
    13561 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
    13621 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
    13681 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
    13741 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
    13801 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
    13861 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
    13921 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
    13981 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
    14041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
    14101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    14161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    14221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    14281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    14341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    14401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    14461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    14521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    14581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    14641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    14701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    14761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    14821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    14881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
    14941 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
    15001 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
    15061 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
    15121 tcacacagga aacagctatg accatgatta cgccaagctc ggaattaacc ctcactaaag
    15181 ggaacaaaag ctg
//
