LOCUS       sequence-398131-       12254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  double-DHFR-degron controlled expression of dCas9-BFP-KRAB from the 
            CLYBL locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12254)
  AUTHORS   Tian R, Gachechiladze MA, Ludwig CH, Laurie MT, Hong JY, Nathaniel 
            D, Prabhu AV, Fernandopulle MS, Patel R, Abshari M, Ward ME, 
            Kampmann M
  TITLE     CRISPR Interference-Based Platform for Multimodal Genetic Screens in
            Human iPSC-Derived Neurons.
  JOURNAL   Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 
            10.1016/j.neuron.2019.07.014.
  PUBMED    31422865
REFERENCE   2  (bases 1 to 12254)
  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: "Neuron. 
            2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 
            10.1016/j.neuron.2019.07.014."
COMMENT     Sequence Label: pRT029
FEATURES             Location/Qualifiers
     source          1..12254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    419..444
                     /label=SA
                     /note="splice acceptor site"
     primer_bind     complement(486..503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    492..716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        722..1101
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1379..2395
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2319..2341
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2403..2422
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2466..2939
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILER"
     regulatory      2958..2967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2964..2990,2991..2993,2994..5480,5481..5483,
                     5484..7067)
                     /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="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             2964..7067
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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             7071..7097
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7077..7103
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             7116..7136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7143..7163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7203..7898
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             7944..8129
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             8130..8177
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8178..8654
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     primer_bind     complement(8744..8763)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8809..8864
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8863..8882)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(10196..10784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10276..10295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10955..11746,11747..11815))
                     /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     11578..11597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttctttggt ggtggtcaca gtcatgcttc agcccagttt ccacttggac aaatggtaca
       61 tattttcaat gagatgaaaa ttaagataca atccatgtgc tcagagagtg atcacagctc
      121 tgactaaaca ctgtgcccca aagtgttgag gaattgggaa aacctagctg agttagtggt
      181 ctcttttctg ttacaataaa gctcataatg aaaattagcc ttctttgttc ttccccaagt
      241 ctttctttct agacgaaact actttcaact gttttaactt ccttactgtt aacttccata
      301 tttctagata gtatggtcag actgttattt cttgactttt aaatgtaaga tattatctac
      361 tgacttcctt ctatgtaaga tgaggattga gctctcttac ccttctccca gcggccgcct
      421 gacctcttct cttcctccca cagggcctcg agagatctgg cagcggatag aaacccgctg
      481 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
      541 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
      601 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
      661 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatgggtct
      721 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      781 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      841 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      901 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      961 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     1021 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     1081 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     1141 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     1201 atgggggcgg gggggggggg gggncgcgcc rggggggggg gggggggggg gggggggggg
     1261 gggggsgrgg sggrgaggkg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     1321 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     1381 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     1441 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     1501 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
     1561 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
     1621 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
     1681 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     1741 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     1801 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     1861 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     1921 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1981 gccgcctcgg gccggggrgg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     2041 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     2101 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     2161 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     2221 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     2281 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     2341 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     2401 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaatgtc gacgctagcg
     2461 ccaccatgat cagtctgatt gcggcgttag cggtagatta cgttatcggc atggaaaacg
     2521 ccatgccgtg gaacctgcct gccgatctcg cctggtttaa acgcaacacc ttaaataaac
     2581 ccgtgattat gggccgccat acctgggaat caatcggtcg tccgttgcca ggacgcaaaa
     2641 atattatcct cagcagtcaa ccgagtacgg acgatcgcgt aacgtgggtg aagtcggtgg
     2701 atgaagccat cgcggcgtgt ggtgacgtac cagaaatcat ggtgattggc ggcggtcgcg
     2761 ttattgaaca gttcttgcca aaagcgcaaa aactgtatct gacgcatatc gacgcagaag
     2821 tggaaggcga cacccatttc ccggattacg agccggatga ctgggaatcg gtattcagcg
     2881 aattccacga tgctgatgcg cagaactctc acagctattg ctttgagatt ctggagcggg
     2941 gctctggagg gactagtgcc accatggaca agaagtacag catcggcctg gccatcggca
     3001 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     3061 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcggc gccctgctgt
     3121 tcgacagcgg agaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     3181 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     3241 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     3301 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     3361 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     3421 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     3481 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     3541 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     3601 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     3661 agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca
     3721 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     3781 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     3841 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     3901 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     3961 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     4021 accagagcaa gaacggctac gccggctaca tcgatggcgg agccagccag gaagagttct
     4081 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     4141 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     4201 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     4261 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     4321 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     4381 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc cagcgcccag agcttcatcg
     4441 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     4501 tgctgtacga gtacttcacc gtgtacaacg agctgaccaa agtgaaatac gtgaccgagg
     4561 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg gacctgctgt
     4621 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     4681 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     4741 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     4801 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     4861 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     4921 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     4981 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     5041 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     5101 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     5161 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     5221 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     5281 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     5341 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     5401 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     5461 tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac gactccatcg
     5521 ataacaaagt gctgactcgg agcgacaaga accggggcaa gagcgacaac gtgccctccg
     5581 aagaggtcgt gaagaagatg aagaactact ggcgccagct gctgaatgcc aagctgatta
     5641 cccagaggaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     5701 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     5761 agatcctgga ctcccggatg aacactaagt acgacgagaa cgacaaactg atccgggaag
     5821 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     5881 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     5941 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     6001 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     6061 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     6121 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaaca ggcgagatcg
     6181 tgtgggataa gggccgggac tttgccaccg tgcggaaagt gctgtctatg ccccaagtga
     6241 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     6301 agaggaacag cgacaagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     6361 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     6421 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     6481 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     6541 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     6601 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     6661 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     6721 agcagaaaca gctgtttgtg gaacagcaca aacactacct ggacgagatc atcgagcaga
     6781 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaag gtgctgagcg
     6841 cctacaacaa gcacagagac aagcctatca gagagcaggc cgagaatatc atccacctgt
     6901 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     6961 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     7021 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacgcc tatccctatg
     7081 acgtgcccga ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag
     7141 atcctaagaa aaagcggaaa gtggacggca ttggtagtgg gagcaacggc agcagcggat
     7201 ccagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
     7261 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     7321 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
     7381 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
     7441 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     7501 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     7561 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     7621 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     7681 ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca
     7741 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     7801 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     7861 actgcgacct ccctagcaaa ctggggcaca agcttaatgg cggtggcgga gggatggatg
     7921 ctaagtcact aactgcctgg tcccggacac tggtgacctt caaggatgta tttgtggact
     7981 tcaccaggga ggagtggaag ctgctggaca ctgctcagca gatcgtgtac agaaatgtga
     8041 tgctggagaa ctataagaac ctggtttcct tgggttatca gcttactaag ccagatgtga
     8101 tcctccggtt ggagaaggga gaagagccca aaaggccggc ggccacgaaa aaggccggcc
     8161 aggcaaaaaa gaaaaagatc agtctgattg cggcgttagc ggtagatcac gttatcggca
     8221 tggaaaccgt catgccgtgg aacctgcctg ccgatctcgc ctggtttaaa cgcaacacct
     8281 taaataaacc cgtgattatg ggccgccata cctgggaatc aatcggtcgt ccgttgccag
     8341 gacgcaaaaa tattatcctc agcagtcaac cgagtacgga cgatcgcgta acgtgggtga
     8401 agtcggtgga tgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg gttattggcg
     8461 gcggtcgcgt ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg acgcatatcg
     8521 acgcagaagt ggaaggcgac acccatttcc cggattacga gccggatgac tgggaatcgg
     8581 tattcagcga attccacgat gctgatgcgc agaactctca cagctattgc tttgagattc
     8641 tggagcggcg ataaacgcgt gaattcactc ctcaggtgca ggctgcctat cagaaggtgg
     8701 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     8761 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     8821 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     8881 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     8941 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     9001 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     9061 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     9121 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     9181 tatggagatc gatatctcct catccttcca acataaatat attttgggat tatatcaaca
     9241 ttcaatgtta cttaaagtga ccttgtaaat attttcacaa ctgagccatg tttgatttgt
     9301 atacttatgt ttactttact gtttttcctg aagttaataa ttgccttgaa tttatttatt
     9361 tctttaaaaa tgtttcatta ctcaggactg tagtttacat tacgattctt tgtgttatac
     9421 agttgatggg tttcttttct ttcttaattt ctttaaaaaa tagagatggg gtcttactat
     9481 attacccagg ctggtcttga agtcctgggc tcaagtgatc ttcctgtctc agcctaccaa
     9541 gtagctgaga ctataggtgc aaaaaagcca ctatacctgg ctagtttaca ggttttaaca
     9601 aatgcattat gccacgtatc cattattaca ggatcacaca agatattttc attaccctga
     9661 aagcatccct gtgttccacc aattcatcct gcctccatga gccgctggca accactgatc
     9721 tctatagttt tgccttttct aaaatgtcat ataattggaa tcatacagtc tgtagcattt
     9781 tcagactagc ttttaaaatt tggcaatatg catttaaggt tcctccttaa atgtgaaggg
     9841 catagccaat gtggcttgat agctcatttc tttttattgg tgaatatttc attgtctgga
     9901 tgttccacag tttgtttatc cattcaccta ttcaatttgc tttttttctg tgtatctatc
     9961 actaattcaa tactggactc tccaacagag ccgtcccgct tccctaaggc cctgctctgg
    10021 gcttctgggt ttgagtcctt ggcaagccca ggagaggcgc tcaggcttcc ctgtccccct
    10081 tcctcgtcca ccatctcatg cccctggctc tcctgcccct tccctacagg ggttcctggc
    10141 tctgctctaa gggcgaattc gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
    10201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
    10261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
    10321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    10381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    10441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    10501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    10561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
    10621 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    10681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    10741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    10801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
    10861 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
    10921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
    10981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
    11041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
    11101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
    11161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
    11221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
    11281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
    11341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
    11401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    11461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    11521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    11581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    11641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    11701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    11761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    11821 tcctttttca atagcaggag aatcatgcca atgggccaat atacattctg acccacagtt
    11881 tcataataaa ataaaatggt tgtggttgta agccactatg tttcagagtg gtttgttaca
    11941 cagcaataaa taactaatat agtaggcata ccatcaagtc caaagtaggt agagaagaat
    12001 gtaaatagca gagcaaaaca gcatgactgg tggctgggag gcttaaaact gggacaggat
    12061 cagagtcatg aaagaagtca aagaaatggt tcagaagtaa ggctgagact gacttacaaa
    12121 agctgaaagt ccctttaagt tggtgtttgg tgcattggca ggggcaggta tggtgactta
    12181 aaagagccat gctcaacaag atcaagcaca acacaatcac gggtcacccc agcagacctt
    12241 agcgagtcta gcca
//
