LOCUS       sequence-355056-       17381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of dCas9-VP64 in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17381)
  AUTHORS   Fischer F, Benner C, Goyala A, Grigolon G, Vitiello D, Wu J, Zarse 
            K, Ewald CY, Ristow M
  TITLE     Ingestion of single guide RNAs induces gene overexpression and 
            extends lifespan in C. elegans via CRISPR activation.
  JOURNAL   J Biol Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085.
  PUBMED    35636511
REFERENCE   2  (bases 1 to 17381)
  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: "J Biol 
            Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085."
COMMENT     Sequence Label: pdestMB14_sur5p-dCas9-VP64
FEATURES             Location/Qualifiers
     source          1..17381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2208..2228
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    4530..4554
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4566..4631
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4638..4658
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4668..5711,5763..6833,6885..7910,7962..8924)
                     /codon_start=1
                     /product="S. pyogenes Cas9, codon optimized for C. elegans 
                     with synthetic introns"
                     /label=Cas9
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8949..8969
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8979..9128
                     /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             9135..9161
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(9173..9197)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9244..9272)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     10020..10040
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     10977..10993
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(12950..12961)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(16740..16759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(16741..16759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(16766..16783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(16766..16782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16775..16797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(16923..17378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(17060..17081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     17272..17291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtacatcaa ctttgtatag
     2221 aaaagttgag tctatacgaa gtatattttt tcttattttg gtttatcgga ggtatgcagt
     2281 tccaaatgtt ggattttcta ccctactaga aatttccttt agttatacaa ttttaattcg
     2341 aagttgtatc ttgaaaagga aaccatcatc aacaacaaat aagcttgaga ctagaattgc
     2401 taatcgtctg caccaaatat gaaattcgtt gaataaatta tataaaacac ttgacaatca
     2461 taaagtattt tacatacata tgtatagtta atgtcatatt agcatattta gtattgactt
     2521 aacacgcaga aataccaaag aaaccctgtt taattcattt ccatgtttta ttcgctttaa
     2581 ctagggaatg accagaataa gtggagcgat attcaaaaaa aaaaaattgt atcggaaagc
     2641 tgacattctc tactataaga atatgactga aatttttgcc cattcgggct ggaaatctga
     2701 atggaaattg tttatatgag ggcatccatg gttgtagtta caattttgag aaaaaagttt
     2761 aaattttagg ggaaaagatg ttgaatgaat cattatggtg ttgctcgaca cattgatcat
     2821 gctgatatcg atcagaaaaa ttaaatttgg gcttttatat agtttcgaaa aaacacgttt
     2881 aaatttttga aaaattttac gttagggtgg aattgaaccc cagagtccca ccatgcttgt
     2941 catccactct atccactcgg ccatctcgct gttgcaggca gtactaattg gggatagtga
     3001 aaagacttaa cgtagaattt cagacgtaaa aatttcagat ttccagcccg aatgggccaa
     3061 aatttcagtc atattcttat agtagagaat gtcagctttc cgatacaata tttttttttt
     3121 gaatatcgct ccacttattc tggtcattcc ctagtaagtt ttttcagttt taccagaaat
     3181 aaaaccactc ataaaaacta taaactacac atttaaactg taaatgttgg aaatcatttt
     3241 gaccatcaaa caatagctat cggaaaatgc aaaggccatt ccgtgatggc acgctcttct
     3301 tctatccatc gctccttttc aacgaaacac acacacaaac gcagactatt ggaaaagaaa
     3361 aatatagaaa aaaccttttt tctccttgtc ccttgtgttg atctcacttt tcagtggacg
     3421 tgtgcagtgt ggaggccaga gactgtgctg accgacggac gcgcaggcgg taaacatacg
     3481 ttgcagtgga gagtgtagtc cctcgtcggc tctctgtgta tttctttcaa tgcctgtgtc
     3541 tcggtctatc tttgtctcta tctgttttaa aagttttttg caaagtttca atttcttttt
     3601 tattttctct tctttttttg cgcatttatt ttttttaaag agcgctgttc ttcaattgga
     3661 aaacagtata acgatcgaaa taaaaaaaaa acttgaaata gtatcggaat tgagatgggg
     3721 gcaattgaaa ttgaagacgc aacaaaaaca tgtttttatc gtctgaaatt atctaaactt
     3781 tgaatctaac gtgtcaatgg ctacttttac agtagacttt ttagagtaat gttaaaaagg
     3841 ataatatagg tttttcttca aactattttg tcacgccgca tatattcttt tcatattcca
     3901 ttcgcaaagt gcatgatgta acaaaaatga aacgtcataa ttttttaatc tttcacactt
     3961 tttttgccgg ccgttaaatc aaaatcaaat caacttaatc caacacgtgc gggcttgcaa
     4021 ttttattaca ccggtgtcct acagcgccat gttatcttct ttgattagag atctctcacg
     4081 caacatttga cattgaaaat tttgtctccg aattgttgtg tacctacttt gtcccgatgt
     4141 cccaaatttt cactgataac aagagatatc tcaactgttc ttatcacgtt gatacgtaga
     4201 acgttttaca ctaaaccaaa ttgcagtccc cttctacgaa ccgtttgcta attgttttgc
     4261 atagaacagt gatcaatgtg ttgtgctcag tagtcaatga tgttcgtctc tcatttttcc
     4321 tacaacttca ctgaaaaggt tttgaatatc tggtgattgt aggcaatcac accacagtta
     4381 atggcctggc aaataatgat aagcgtgtga taaacctcac aaaaatacgc ctagaagtgg
     4441 tgatatcgct agttttgtgt cctttcgatt tttttcgtgt tgcttttttc tgtggataaa
     4501 cctttgaact ttacattttg ttttcagaga caagtttgta caaaaaagca ggctgcggcc
     4561 gcatggatta taaagatcat gatggagatt ataaagatca tgatattgat tataaagatg
     4621 atgatgataa aatggctcca aaaaaaaaaa gaaaagttgg aagaggaatg gacaaaaaat
     4681 acagcatcgg ccttgctatc ggaacgaact ccgtcggatg ggccgtcatc accgacgagt
     4741 acaaggtccc atccaagaag ttcaaggtcc tcggaaacac cgaccgtcac tccatcaaga
     4801 agaacctcat cggagccctc ctcttcgact ccggagagac cgccgaggcc acccgtctca
     4861 agcgtaccgc ccgtcgtcgt tacacccgtc gtaagaaccg tatctgctac ctccaagaga
     4921 tcttctccaa cgagatggcc aaggtcgacg actccttctt ccaccgtctc gaggagtcct
     4981 tcctcgtcga ggaggacaag aagcacgagc gtcacccaat cttcggaaac atcgtcgacg
     5041 aggtcgccta ccacgagaag tacccaacca tctaccacct ccgtaagaag ctcgtcgact
     5101 ccaccgacaa ggccgacctc cgtctcatct acctcgccct cgcccacatg atcaagttcc
     5161 gtggacactt cctcatcgag ggagacctca acccagacaa ctccgacgtc gacaagctct
     5221 tcatccaact cgtccaaacc tacaaccaac tcttcgagga gaacccaatc aacgcctccg
     5281 gagtcgacgc caaggccatc ctctccgccc gtctctccaa gtcccgtcgt ctcgagaacc
     5341 tcatcgccca actcccagga gagaagaaga acggactctt cggaaacctc atcgccctct
     5401 ccctcggact caccccaaac ttcaagtcca acttcgacct cgccgaggac gccaagctcc
     5461 aactctccaa ggacacctac gacgacgacc tcgacaacct cctcgcccaa atcggagacc
     5521 aatacgccga cctcttcctc gccgccaaga acctctccga cgccatcctc ctctccgaca
     5581 tcctccgtgt caacaccgag atcaccaagg ccccactctc cgcctccatg atcaagcgtt
     5641 acgacgagca ccaccaagac ctcaccctcc tcaaggccct cgtccgtcaa caactcccag
     5701 agaagtacaa ggtaagttta aacatatata tactaactaa ccctgattat ttaaattttc
     5761 aggagatctt cttcgaccaa tccaagaacg gatacgccgg atacatcgac ggaggagcct
     5821 cccaagagga gttctacaag ttcatcaagc caatcctcga gaagatggac ggaaccgagg
     5881 agctcctcgt caagctcaac cgtgaggacc tcctccgtaa gcaacgtacc ttcgacaacg
     5941 gatccatccc acaccaaatc cacctcggag agctccacgc catcctccgt cgtcaagagg
     6001 acttctaccc attcctcaag gacaaccgtg agaagatcga gaagatcctc accttccgta
     6061 tcccatacta cgtcggacca ctcgcccgtg gaaactcccg tttcgcctgg atgacccgta
     6121 agtccgagga gaccatcacc ccatggaact tcgaggaggt cgtcgacaag ggagcctccg
     6181 cccaatcctt catcgagcgt atgaccaact tcgacaagaa cctcccaaac gagaaggtcc
     6241 tcccaaagca ctccctcctc tacgagtact tcaccgtcta caacgagctc accaaggtca
     6301 agtacgtcac cgagggaatg cgtaagccag ccttcctctc cggagagcaa aagaaggcca
     6361 tcgtcgacct cctcttcaag accaaccgta aggtcaccgt caagcaactc aaggaggact
     6421 acttcaagaa gatcgagtgc ttcgactccg tcgagatctc cggagtcgag gaccgtttca
     6481 acgcctccct cggaacctac cacgacctcc tcaagatcat caaggacaag gacttcctcg
     6541 acaacgagga gaacgaggac atcctcgagg acatcgtcct caccctcacc ctcttcgagg
     6601 accgtgagat gatcgaggag cgtctcaaga cctacgccca cctcttcgac gacaaggtca
     6661 tgaagcagct taagcgtcgt cgttacaccg gatggggacg tctctcccgt aagctcatca
     6721 acggaatccg tgacaagcaa tccggaaaga ccatcctcga cttcctcaag tccgacggat
     6781 tcgccaaccg taacttcatg caactcatcc acgacgactc cctcaccttc aaggtaagtt
     6841 taaacagttc ggtactaact aaccatacat atttaaattt tcaggaggac atccaaaagg
     6901 cccaagtctc cggacaagga gactccctcc acgagcacat cgccaacctc gccggatccc
     6961 cagccatcaa gaagggaatc ctccaaaccg tcaaggtcgt cgacgagctc gtcaaggtca
     7021 tgggacgtca caagccagag aacatcgtca tcgagatggc ccgtgagaac caaaccaccc
     7081 aaaagggaca aaagaactcc cgtgagcgta tgaagcgtat cgaggaggga atcaaggagc
     7141 tcggatccca aatcctcaag gagcacccag tcgagaacac ccaactccaa aacgagaagc
     7201 tctacctcta ctacctccaa aacggacgtg acatgtacgt cgaccaagag ctcgacatca
     7261 accgtctctc cgactacgac gtcgacgcta tcgtcccaca atccttcctc aaggacgact
     7321 ccatcgacaa caaggtcctc acccgttccg acaagaaccg tggaaagtcc gacaacgtcc
     7381 catccgagga ggtcgtcaag aagatgaaga actactggcg tcaactcctc aacgccaagc
     7441 tcatcaccca acgtaagttc gacaacctca ccaaggctga gcgtggagga ctctccgagc
     7501 tcgacaaggc cggattcatc aagcgtcaac tcgtcgagac ccgtcaaatc accaagcacg
     7561 tcgcccaaat cctcgactcc cgtatgaaca ccaagtacga cgagaacgac aagctcatcc
     7621 gtgaggtcaa ggtcatcacc ctcaagtcca agctcgtctc cgacttccgt aaggacttcc
     7681 aattctacaa ggtccgtgag atcaacaact accaccacgc ccacgacgcc tacctcaacg
     7741 ccgtagtagg aaccgccctc atcaagaagt acccaaagct cgagtccgag ttcgtctacg
     7801 gagactacaa ggtctacgac gtccgtaaga tgatcgccaa gtccgagcaa gagatcggaa
     7861 aggccaccgc caagtacttc ttctactcca acatcatgaa cttcttcaag gtaagtttaa
     7921 acatgatttt actaactaac taatctgatt taaattttca gaccgagatc accctcgcca
     7981 acggagagat ccgtaagcgt ccactcatcg agaccaacgg agagaccgga gagatcgtct
     8041 gggacaaggg acgtgacttc gccacagtcc gtaaggtcct ctccatgcca caagtcaaca
     8101 tcgtcaagaa gaccgaggtc caaaccggag gattctccaa ggagtccatc ctcccaaagc
     8161 gtaactccga caagctcatc gcccgtaaga aggactggga cccaaagaag tacggaggat
     8221 tcgactcccc aaccgtcgcc tactccgtcc tcgtcgtcgc caaggtcgag aagggaaagt
     8281 ccaagaagct caagtccgtc aaggagctcc tcggaatcac catcatggag cgttcctcct
     8341 tcgagaagaa cccaatcgac ttcctcgagg ccaagggata caaggaggtc aagaaggacc
     8401 tcatcatcaa gctcccaaag tactccctct tcgagctcga gaacggacgt aagcgtatgc
     8461 tcgcctccgc cggagagctc caaaagggaa acgagctcgc cctcccatcc aagtacgtca
     8521 acttcctcta cctcgcctcc cactacgaga agctcaaggg atccccagag gacaacgagc
     8581 aaaagcaact cttcgtcgag caacacaagc actacctcga cgagatcatc gagcaaatct
     8641 ccgagttctc caagcgtgtc atcctcgccg acgccaacct cgacaaggtc ctctccgcct
     8701 acaacaagca ccgtgacaag ccaatccgtg agcaagccga gaacatcatc cacctcttca
     8761 ccctcaccaa cctcggagcc ccagccgcct tcaagtactt cgacaccacc atcgaccgta
     8821 agcgttacac ctccaccaag gaggtcctcg acgccaccct catccaccaa tccatcaccg
     8881 gactctacga gacccgtatc gacctctccc aactcggagg agacccaatt gctggatcaa
     8941 aagcttcacc aaaaaaaaaa agaaaagttt ctagagctga tgctcttgat gatttcgatc
     9001 ttgatatgct tggatcagat gctcttgatg atttcgatct tgatatgctt ggatcagatg
     9061 ctcttgatga tttcgatctt gatatgcttg gatcagatgc tcttgatgat ttcgatcttg
     9121 atatgcttat taattatcca tatgatgttc cagattatgc ttcatagggt acccccagct
     9181 ttcttgtaca aagttgatgt accggtagaa aaaatgagta aaggagaaga acttttcact
     9241 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     9301 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
     9361 actggaaaac tacctgttcc atgggtaagt ttaaacatat atatactaac taaccctgat
     9421 tatttaaatt ttcagccaac acttgtcact actttctgtt atggtgttca atgcttctcg
     9481 agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     9541 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg taagtttaaa
     9601 cagttcggta ctaactaacc atacatattt aaattttcag gtgctgaagt caagtttgaa
     9661 ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac
     9721 attcttggac acaaattgga atacaactat aactcacaca atgtatacat catggcagac
     9781 aaacaaaaga atggaatcaa agttgtaagt ttaaacatga ttttactaac taactaatct
     9841 gatttaaatt ttcagaactt caaaattaga cacaacattg aagatggaag cgttcaacta
     9901 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     9961 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
    10021 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
    10081 tagcattcgt agaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc
    10141 aaaaataata ggggccgctg tcatcagagt aagtttaaac tgagttctac taactaacga
    10201 gtaatattta aattttcagc atctcgcgcc cgtgcctctg acttctaagt ccaattactc
    10261 ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca
    10321 aaaaacttct cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa
    10381 ttgtgtagat tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc
    10441 tccccccatt aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta
    10501 tgttttttac ttctgataaa tttttttgaa acatcataga aaaaaccgca cacaaaatac
    10561 cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc
    10621 ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt
    10681 caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg gtttccccta
    10741 ttgtttgtca agatttcgag gacggcgttt ttcttgctaa aatcacaagt attgatgagc
    10801 acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa
    10861 gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg acagaataca
    10921 ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc ccccctcgag
    10981 gtcgacggta tcgataagct tcagtaaaag aagtagaatt ttatagtttt ttttctgttt
    11041 gaaaaattct ccccatcaat gttctttcaa ataaatacat cactaatgca aagtattcta
    11101 taacctcata tctaaattct tcaaaatctt aacatatctt atcattgctt taagtcaaac
    11161 gtaacattaa aaaaaatgtt ttggaaaatg tgtcaagtct ctcaaaattc agttttttaa
    11221 accactccta tagtcctata gtcctatagt tacccatgaa atccttatat attactgtaa
    11281 aatgtttcaa aaaccattgg caaattgcca gaactgaaaa tttccggcaa attggggaac
    11341 cggcaaattg ccaatttgct gaatttgccg gaaaaacggt aattgccgaa agtttttgac
    11401 acgaaaatgg caaattgtgg ttttaaaatt tttttttttg gaaatttcag aatttcaatt
    11461 ttaatcggca aaactgtagg catcctaaga atgttcctac atctattttg aaaagtaagc
    11521 gaattctatg aaaatgtcta aagaaaatgg ggaaacaatt tcaaaaaggc acagtttcaa
    11581 gtgtttccga attatactaa atccctctaa aaacttccgg caaattgata tccgtaaaag
    11641 agcaaatccg catttttgcc gaaaattaaa atttccgaca aatcggcaaa ccggcaattt
    11701 ggcgaaattt gccggaaggc aattgccgcc cacccctgtt ccagaggttc aaactggtag
    11761 caaagctcaa aatttctcaa attctccaat ttttttttga attttggcag tgtaccaaaa
    11821 tgacattcag tcatattggt ttattataga tttatttaga taaaatccta aatgattcta
    11881 cctttaaaga tgcccacttt aaaagtaatg actcaaactt caaattgctc taagattcta
    11941 ttgaattacc atcttttcct ctcattttct ctcactgtct atttcatcac aaattcatcc
    12001 ctctctccct ctctttctct ctccctctct ctctttctct ttgctcatca tctgtcattt
    12061 tgtccgttcc tctctctgcg ccctcagcgt tccccacact ctctcgcttc tcttttccta
    12121 gacgtcttct tttttcatct tcttcagccc tttttcgcca ttttccatct ctgtcaatca
    12181 ttacggacga cccccattaa tttttggatg tgctctgcga gcaagagcac acggaactca
    12241 tttatttttt gttggaattt ctttttcatt ttagttagtt aaaaggttat tcagtgcaaa
    12301 aattgatttt ttgctctttt ggtacattag aggggtttga aagttttcca taaaaagggt
    12361 ttccggagta aaaaatctaa aaatttttac aaacatttcc aaaaattttg ggaacatatc
    12421 ttttcaaaaa ctcaaaaaaa aacgcaaaca tcataaattt aatcaattaa ttggatgcgt
    12481 tcaaatattt taaaaatatt tttgaattgc atatttcttt actcataaaa aaattattaa
    12541 tgtttaagta aaaaaaatat ttcaggattt taaacaaaaa ctgtaaatcc cataaaacag
    12601 tgttattttt tagttatttt tagtaattat ttgtgaatta aaaacacatt ttttttcaaa
    12661 atattccttg aataaataaa attgaaagtg aattcaaaat atttcaccaa ttttacaaaa
    12721 tttgaaaatt tgaaaaaata ttttctacca aattgtgctc aaaaaatatt attaagttcc
    12781 caggaaattt atgatgaaaa cttggaatat ttttggtaaa attttctcaa aaaattgaaa
    12841 gttgtcaaaa acacactaac actatacctc gcggcataga aaaaactggt ggccgaattt
    12901 tttaaaccaa tttaaattaa atatattttt gcactttgta caaaaaaatt tgaagttccc
    12961 aggaaattca tacggaaata ttttccagaa cttttataat ttttatttta attctcaaaa
    13021 tttacaataa aaacttggaa acctgaaatt tgcttaaaga ataaccctat cccccacggc
    13081 ctagaaaata ctggtggccg aattttttcc gcagccacac cacctctaat ctccagagca
    13141 ctctccaaaa ttcccataat ccccaaaatt ttcagctaca acagcatata tgaaggcaga
    13201 tctacaacaa caatcgatcg cacccggctc ggcaaccttc ccgtctcagg tgaggaagac
    13261 gcagagaaat gagagagaga gcgtgtttat gcacgcagta tactaactcc tgatatccaa
    13321 tttcatatat cgatatgctt gctttctttt cttctacgtt gaaattgttt gaacttttct
    13381 caaaaaacgc ggattttaga gagacttaga gttgctccaa ataaatttaa tttgcaattt
    13441 tgcacttact attagaatct acttttatag cgctaaaatg accgaataac acagtagatt
    13501 ttaaaagaaa attttaaaaa ttattttttt gtgcagtcaa atttcgctag tttttgccac
    13561 tcatttttga acatttgtat atgtgccttt cggtggttca atcaggggtg gcacgtggtg
    13621 tcagagtgtt ccatttcggt ttgatctacg ttgatctaca aaaaatgcgg gagaagagac
    13681 gcagagttct caactgattt cacatggtta tgaaccaggg gtgttcggca aatttgccgt
    13741 ttgccgagct gggcaaattt tgagatttgc cgcacacccc ttttatgaac gtgctgacgt
    13801 cacatctttt tggacaaaga attcccgcat tttttgtata tcaaaccgtg atgggacagc
    13861 ctggcaccac gtggggttgg cggcaaatga ttttttccga caaatcggca aattggcaaa
    13921 ttgccggaat ttaaagtttc tggcaaattg ttgatttgcc gaaaaaaaat ttcaaaaagg
    13981 cacagtttta agtgtttccg tcttataaaa tatttctcta aaaatttccg gaaaacggaa
    14041 aattgccgga aatacaaagt ttcgtcaaat cggcaaacct gcaatttgcc gaacggcaac
    14101 tgccgccaac acctggttcc caaaaaagcg tgcactgttt gaatattaaa aaaaaaattc
    14161 acacaaaaat aataaattga aataggctaa attcacatct atgtacaaaa attgcatgcc
    14221 agcaccggtc aattttctca attacttttt gtctattttt gccgaacata atttgctgtt
    14281 tccttggcag aatacagaaa actctttttt ttttcagatg ccccgacccc caccggtaac
    14341 cgaacaggct ataaccaccg aggcggagct tctcgcgaaa aatcaaatta caccaaatga
    14401 tgtgctagct cttccaggaa tcactcaagg taagtgaata tttgctctat tgataaacgc
    14461 gccgtgtact ccacgtggac aaacatatca aattttcagg attcctatgc tccccatcgg
    14521 caaacgtcta taacattgag ttcaccaaat tccaaatccg tgacctcgac acggaacacg
    14581 tgctcttcga aattgcaaaa ccggaaaatg agacggaaga gaatctgcag gcgcaagccg
    14641 aatcggcaag atatgtccga tatcgatttg cgccgaattt tctgaaatta aagacggtcg
    14701 gcgcgacggt cgagttcaag gtcggcgatg tgccgatcac acattttcga atgattgaac
    14761 ggcacttttt caaggatcgt ttattgaaat gctttgattt tgaatttgga ttctgtatgc
    14821 caaattcgag aaacaactgt gaacatatct atgaatttcc acaactttca caacaactca
    14881 gtgagtttga agcgggaaaa attatttgag ctcattttgt caatagagcg aattttctag
    14941 gtttttaaaa ataaaataaa tattgacaaa aagctgccct acagtagttc aaaatgtgaa
    15001 aacttggagg aaattcgagt tttctctgag aagtttttgg cgggtaattc aaattttctg
    15061 agaaaaaaat tagccgcgag aattcaaatt ttctgagaaa attatttgac gggaaattca
    15121 aaatttctgg aaatgtattg aacatttgag aaattttgtt aaaaactaca ttaaaccgat
    15181 aaaaaagttc gcaaattttt tttaagtgct catggtgcat cgaagagaag aattcttgcc
    15241 aaaactagcc catgaaatta aaaattatcg cggaaaaaaa ctgactaatt ggaaagtaaa
    15301 atgaaatttg caaaaaaaaa atatgttttt ttgttgtaaa attaaaaaaa aaactgtata
    15361 ttttcagtgg acgacatgat caataatcca aacgagaccc gctccgatag cttctatttt
    15421 gtggagaata agctcgtaat gcacaataag gccgactact cgtatgatgc ataatttccc
    15481 gccgatggaa taaaactaat aatttttctc gcaatttata gaaatcatgt gattttccta
    15541 ccgctaaatt taaaatttcc taatttttca ctatatttct cacaaaatgc attccgaaaa
    15601 ctctcatttt tcagcttttc aatacaaatc ccaataaatg atttttcaaa tataatgttc
    15661 tcaattgata tgttttattt gaatgttgga aaaaaccata aaattaaaaa aaacttgaat
    15721 tattcaaaaa taattcggcc cttttacaaa aaattatcat aaaaattaaa tttggaaaat
    15781 ctgtgatttg tcgctttgcc ggaagtttcc aattccggca actaaccgat ttgccgatta
    15841 gctagaaatt tcaattccga caatgtgccg ctttgccgat ttgccggaaa tttccaattc
    15901 cggcaattaa cggacttgcc gattagctag aaatttaact ttagataaaa cttctcaaca
    15961 aaatcaaatt ggaaagtcag cgatttgccg ctttgccgga agttttcagt tccggcatct
    16021 aaccgatttg ccgatttgcc aaaaatgtca attccgaaaa actgccgatt tgccgctttg
    16081 tcggaaattt caattccgac aaattgccga tttgccgatt tgccgaacat tccaattccg
    16141 accatttgtc gatttgccgg aagttaccaa ttccggcaac taacggattt gccgatttgc
    16201 cggaaaaaaa ttgtttgtcg ccaaccctga tttaaatgac taatcatgtt gcaaaagata
    16261 actgcttgca tcaaaaatcc gatcattggc attcccaaca tcttcttcat ccaaaacacc
    16321 acccttttct tccactaatc atgaattccc gaccaattgt tatcagtagg aaaagcctcc
    16381 ttttatctca ttatcagcaa ttgccttctt cttcttcttc tttgaatacc atttgaatgg
    16441 agatatcatt tacataaatc tgccgccgag attgtgaagc aggaggaaaa gctgaagata
    16501 ctgatattat atgtatattt cattattctt tttccacaaa attttccgac atttaattcc
    16561 acttcacttt catcgtctct ttttccccca ttgtcgcccc cactcatttg tcttcttctt
    16621 caaatacggg tcaatcactt tccgtttttt attcattttt tgttatgttc ggcttaacaa
    16681 atttgaagta ttgttaaaat tctctagagc ggccgccacc gcggtggagc tccaattcgc
    16741 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
    16801 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
    16861 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
    16921 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
    16981 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
    17041 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
    17101 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
    17161 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
    17221 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
    17281 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
    17341 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca g
//
