LOCUS       sequence-160892-       15460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AND gate ABA/GA-inducible VPR-Sp dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15460)
  AUTHORS   Gao Y, Xiong X, Wong S, Charles EJ, Lim WA, Qi LS
  TITLE     Complex transcriptional modulation with orthogonal and inducible 
            dCas9 regulators.
  JOURNAL   Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042.
  PUBMED    27776111
REFERENCE   2  (bases 1 to 15460)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042."
COMMENT     Sequence Label: pSLQ2816 pPB: CAG-PYL1-VPR-p2A-GID1-ABI-WPRE
            P...
FEATURES             Location/Qualifiers
     source          1..15460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..284
                     /label=chicken beta-actin promoter
     intron          286..1303
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1227..1249
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1311..1330
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1358..1367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1940..2089
                     /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             2096..2116
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2546..2917
                     /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
                     EDFSSIADMDFSALLSQISS"
     CDS             2936..3505
                     /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"
     CDS             3515..3571
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    5571..6159
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5624..5644)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6042..6053)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6180..6679
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6215..6238)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6612..6631
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6721..6738
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7012..7710
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      7726..7735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7732..7758,7759..7761,7762..10248,10249..10251,
                     10252..11835)
                     /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             7732..11835
                     /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             11839..11865
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11845..11871
                     /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             11884..11904
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11911..11931
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   12148..12210
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12261..12281)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12261..12279)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12300..12316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12300..12316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12313..12335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12324..12340
                     /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)."
     promoter        complement(join(12348..12354,12355..12372,12373..12378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12393..12414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12531..12548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12702..13290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12782..12801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13461..14252,14253..14321))
                     /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     14084..14103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14322..14426)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14452..14907)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14589..14610)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14801..14820
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15034..15056
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15048..15065
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15049..15065
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15075..15094
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15075..15093
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   15123..15157
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
       61 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      121 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      181 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      241 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg
      301 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      361 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      421 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      481 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      541 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      601 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
      661 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
      721 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
      781 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
      841 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
      901 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
      961 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1021 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1081 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1141 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1201 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1261 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1321 ggttattgtg ctgtctcatc attttggcaa agaattcgcc accatgggtg ggggcgcgcc
     1381 aactcaagac gaattcaccc aactctccca atcaatcgcc gagttccaca cgtaccaact
     1441 cggtaacggc cgttgctcat ctctcctagc tcagcgaatc cacgcgccgc cggaaacagt
     1501 atggtccgtg gtgagacgtt tcgataggcc acagatttac aaacacttca tcaaaagctg
     1561 taacgtgagt gaagatttcg agatgcgagt gggatgcacg cgcgacgtga acgtgataag
     1621 tggattaccg gcgaatacgt ctcgagagag attagatctg ttggacgatg atcggagagt
     1681 gactgggttt agtataaccg gtggtgaaca taggctgagg aattataaat cggttacgac
     1741 ggttcataga tttgagaaag aagaagaaga agaaaggatc tggaccgttg ttttggaatc
     1801 ttatgttgtt gatgtaccgg aaggtaattc ggaggaagat acgagattgt ttgctgatac
     1861 ggttattaga ttgaatcttc agaaacttgc ttcgatcact gaagctatga acgaattctc
     1921 aagcgctgca ggtactagtg atgctttaga cgattttgac ttagatatgc ttggttcaga
     1981 cgcgttagac gacttcgacc tagacatgtt aggctcagat gcattggacg acttcgattt
     2041 agatatgttg ggctccgatg ccctagatga ctttgatcta gatatgctag gtagtcccaa
     2101 aaagaagagg aaagtgggat cccagtatct gcccgacaca gatgatagac accgaatcga
     2161 agagaaacgc aagcgaacgt atgaaacctt caaatcgatc atgaagaaat cgcccttctc
     2221 gggtccgacc gatcccaggc ccccaccgag aaggattgcg gtcccgtccc gctcgtcggc
     2281 cagcgtgccg aagcctgcgc cgcagcccta ccccttcacg tcgagcctga gcacaatcaa
     2341 ttatgacgag ttcccgacga tggtgttccc ctcgggacaa atctcacaag cctcggcgct
     2401 cgcaccagcg cctccccaag tccttccgca agcgcctgcc ccagcgcctg caccggcaat
     2461 ggtgtccgcc ctcgcacagg cccctgcgcc cgtccccgtg ctcgcgcctg gaccgcccca
     2521 ggcggtcgct ccaccggctc cgaagccgac gcaggccgga gagggaacac tctccgaagc
     2581 acttcttcaa ctccagtttg atgacgagga tcttggagca ctccttggaa actcgacaga
     2641 ccctgcggtg tttaccgacc tcgcgtcagt agataactcc gaatttcagc agcttttgaa
     2701 ccagggtatc ccggtcgcgc cacatacaac ggagcccatg ttgatggaat accccgaagc
     2761 aatcacgaga cttgtgacgg gagcgcagcg gcctcccgat cccgcacccg cacctttggg
     2821 ggcacctggc ctccctaacg gacttttgag cggcgacgag gatttctcct ccatcgccga
     2881 tatggatttc tcagccttgc tgtcacagat ttccagcggc tctggcagcg gcagccggga
     2941 ttccagggaa gggatgtttt tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt
     3001 tgagggccgc gaggtgtgcc agccaaaacg aatccggcca tttcatcctc caggaagtcc
     3061 atgggccaac cgcccactcc ccgccagcct cgcaccaaca ccaaccggtc cagtacatga
     3121 gccagtcggg tcactgaccc cggcaccagt ccctcagcca ctggatccag cgcccgcagt
     3181 gactcccgag gccagtcacc tgttggagga tcccgatgaa gagacgagcc aggctgtcaa
     3241 agcccttcgg gagatggccg atactgtgat tccccagaag gaagaggctg caatctgtgg
     3301 ccaaatggac ctttcccatc cgcccccaag gggccatctg gatgagctga caaccacact
     3361 tgagtccatg accgaggatc tgaacctgga ctcacccctg accccggaat tgaacgagat
     3421 tctggatacc ttcctgaacg acgagtgcct cttgcatgcc atgcatatca gcacaggact
     3481 gtccatcttc gacacatctc tgtttggaag cggagctact aacttcagcc tgctgaagca
     3541 ggctggagac gtggaggaga accctggacc tcctaggatg gctgcgagcg atgaagttaa
     3601 tcttattgag agcagaacag tggttcctct caatacatgg gttttaatat ccaacttcaa
     3661 agtagcctac aatatccttc gtcgccctga tggaaccttt aaccgacact tagctgagta
     3721 tctagaccgt aaagtcactg caaacgccaa tccggttgat ggggttttct cgttcgatgt
     3781 cttgattgat cgcaggatca atcttctaag cagagtctat agaccagctt atgcagatca
     3841 agagcaacct cctagtattt tagatctcga gaagcctgtt gatggcgaca ttgtccctgt
     3901 tatattgttc ttccatggag gtagctttgc tcattcttct gcaaacagtg ccatctacga
     3961 tactctttgt cgcaggcttg ttggtttgtg caagtgtgtt gttgtctctg tgaattatcg
     4021 gcgtgcacca gagaatccat acccttgtgc ttatgatgat ggttggattg ctcttaattg
     4081 ggttaactcg agatcttggc ttaaatccaa gaaagactca aaggtccata ttttcttggc
     4141 tggtgatagc tctggaggta acatcgcgca taatgtggct ttaagagcgg gtgaatcggg
     4201 aatcgatgtt ttggggaaca ttctgctgaa tcctatgttt ggtgggaatg agagaacgga
     4261 gtctgagaaa agtttggatg ggaaatactt tgtgacggtt agagaccgcg attggtactg
     4321 gaaagcgttt ttacccgagg gagaagatag agagcatcca gcgtgtaatc cgtttagccc
     4381 gagagggaaa agcttagaag gagtgagttt ccccaagagt cttgtggttg tcgcgggttt
     4441 ggatttgatt agagattggc agttggcata cgcggaaggg ctcaagaaag cgggtcaaga
     4501 ggttaagctt atgcatttag agaaagcaac tgttgggttt tacctcttgc ctaataacaa
     4561 tcatttccat aatgttatgg atgagatttc ggcgtttgta aacgcggaat gtcaattggg
     4621 tggaggctct ggaacgcgtg tgcctttgta tggttttact tcgatttgtg gaagaagacc
     4681 tgagatggaa gctgctgttt cgactatacc aagattcctt caatcttcct ctggttcgat
     4741 gttagatggt cggtttgatc ctcaatccgc cgctcatttc ttcggtgttt acgacggcca
     4801 tggcggttct caggtagcga actattgtag agagaggatg catttggctt tggcggagga
     4861 gatagctaag gagaaaccga tgctctgcga tggtgatacg tggctggaga agtggaagaa
     4921 agctcttttc aactcgttcc tgagagttga ctcggagatt gagtcagttg cgccggagac
     4981 ggttgggtca acgtcggtgg ttgccgttgt tttcccgtct cacatcttcg tcgctaactg
     5041 cggtgactct agagccgttc tttgccgcgg caaaactgca cttccattat ccgttgacca
     5101 taaaccggat agagaagatg aagctgcgag gattgaagcc gcaggaggga aagtgattca
     5161 gtggaatgga gctcgtgttt tcggtgttct cgccatgtcg agatccattg gcgatagata
     5221 cttgaaacca tccatcattc ctgatccgga agtgacggct gtgaagagag taaaagaaga
     5281 tgattgtctg attttggcga gtgacggggt ttgggatgta atgacggatg aagaagcgtg
     5341 tgagatggca aggaagcgga ttctcttgtg gcacaagaaa aacgcggtgg ctggggatgc
     5401 atcgttgctc gcggatgagc ggagaaagga agggaaagat cctgcggcga tgtccgcggc
     5461 tgagtatttg tcaaagctgg cgatacagag aggaagcaaa gacaacataa gtgtggtggt
     5521 ggttgatttg aaggtttaaa ccatgcaagc ttgatatcaa gcttatcgat aatcaacctc
     5581 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5641 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5701 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5761 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5821 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5881 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5941 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     6001 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     6061 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     6121 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcctgcaggt aattctaccg
     6181 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     6241 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     6301 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     6361 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     6421 tctcactagg ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     6481 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     6541 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     6601 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     6661 ctcatctccg ggcctttcga cctgcagccc aagcttgtcg acgccaccat gaagagagat
     6721 catcatcatc atcatcatca agataagaag actatgatga tgaatgaaga agacgacggt
     6781 aacggcatgg atgagcttct agctgttctt ggttacaagg ttaggtcatc cgaaatggct
     6841 gatgttgctc agaaactcga gcagcttgaa gttatgatgt ctaatgttca agaagacgat
     6901 ctttctcaac tcgctactga gactgttcac tataatccgg cggagcttta cacgtggctt
     6961 gattctatgc tcaccgacct taatcatatg tcaagcgctg caggtgctac catgagcgag
     7021 ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgga caaccatcac
     7081 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc
     7141 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac tagcttcctc
     7201 tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt caagcagtcc
     7261 ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc
     7321 gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
     7381 gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccttc
     7441 accgagacgc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat ggccctgaag
     7501 ctcgtgggcg ggagccatct gatcgcaaac atcaagacca catatagatc caagaaaccc
     7561 gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga aagaatcaag
     7621 gaggccaaca acgagaccta cgtcgagcag cacgaggtgg cagtggccag atactgcgac
     7681 ctccctagca aactggggca caagcttaat tcaagcgctg caggtgccac catggacaag
     7741 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
     7801 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     7861 aagaagaacc tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg
     7921 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     7981 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     8041 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     8101 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     8161 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     8221 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     8281 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     8341 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     8401 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
     8461 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     8521 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     8581 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     8641 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     8701 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     8761 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc
     8821 gatggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     8881 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     8941 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     9001 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     9061 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     9121 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     9181 aagggcgcca gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     9241 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag
     9301 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     9361 cagaaaaaag ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     9421 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     9481 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     9541 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     9601 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     9661 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     9721 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     9781 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     9841 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     9901 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     9961 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
    10021 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
    10081 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
    10141 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
    10201 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct
    10261 cagagctttc tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac
    10321 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
    10381 cgccagctgc tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc
    10441 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
    10501 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
    10561 gacgagaacg acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
    10621 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
    10681 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
    10741 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
    10801 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
    10861 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
    10921 gagacaaacg gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg
    10981 cggaaagtgc tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
    11041 ggcttcagca aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag
    11101 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
    11161 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
    11221 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
    11281 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
    11341 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
    11401 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
    11461 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa
    11521 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
    11581 gacgctaatc tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga
    11641 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
    11701 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
    11761 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
    11821 cagctgggag gcgacgccta tccctatgac gtgcccgatt atgccagcct gggcagcggc
    11881 tcccccaaga aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacggcatt
    11941 ggtagtggga gcaacggatg actcgagcta gttaaaagtt ttgttacttt atagaagaaa
    12001 ttttgagttt ttgttttttt ttaataaata aataaacata aataaattgt ttgttgaatt
    12061 tattattagt atgtaagtgt aaatataata aaacttaata tctattcaaa ttaataaata
    12121 aacctcgata tacagaccga taaaacacat gcgtcaattt tacgcatgat tatctttaac
    12181 gtacgtcaca atatgattat ctttctaggg ttaatctagt atacgcgtat gcggccgctt
    12241 aattaatcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
    12301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    12361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    12421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    12481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    12541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    12601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    12661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    12721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    12781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    12841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    12901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    12961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    13021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    13081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    13141 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    13201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    13261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    13321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    13381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    13441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    13501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    13561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    13621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    13681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    13741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    13801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    13861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    13921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    13981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    14041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    14101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    14161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    14221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    14281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    14341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    14401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
    14461 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
    14521 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
    14581 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
    14641 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
    14701 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
    14761 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
    14821 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
    14881 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
    14941 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
    15001 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
    15061 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggggcgcgcc attctagatt
    15121 aaccctagaa agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc
    15181 tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc
    15241 tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc
    15301 gtgagtcaaa atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat
    15361 aattatattg ttatttcatg ttctacttac gtgataactt attatatata tattttcttg
    15421 ttatagatat caactagaat gctagcatgg gcccagatct
//
