LOCUS       sequence-214274-       13966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a VPR transcriptional activator fused to the 
            nuclease-deactivated S. pyogenes Cas9, which is governed by an 
            inserted StaPL(TI) module, in mammalian cells. [TI = telaprevir 
            inhibited]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13966)
  AUTHORS   Jacobs CL, Badiee RK, Lin MZ
  TITLE     StaPLs: versatile genetically encoded modules for engineering 
            drug-inducible proteins.
  JOURNAL   Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z.
            Epub 2018 Jul 2.
  PUBMED    29967496
REFERENCE   2  (bases 1 to 13966)
  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"; doi: 
            "10.1038/s41592-018-0041-z"; journalName: "Nat Methods"; date: 
            "2018-07"; volume: "15"; issue: "7"; pages: "523-526"
COMMENT     Sequence Label: pPB-VPR-dSpCas9[StaPL(TI)]
FEATURES             Location/Qualifiers
     source          1..13966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..299
                     /label=chicken beta-actin promoter
     intron          300..1316
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1240..1262
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1324..1343
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1383..1982
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1383..1402)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1879..1899
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2020..2608
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2073..2093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2491..2502)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2629..3128
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2664..2687)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3061..3080
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3152..3161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3161..3310
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             3767..4138
                     /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             4157..4726
                     /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             4754..5452
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      5468..5477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5474..7192
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             9239..9265
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10361..10387
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10367..10393
                     /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             10406..10426
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10433..10453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   10670..10732
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10783..10803)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10783..10801)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10822..10838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10822..10838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10835..10857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10846..10862
                     /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(10870..10876,10877..10894,10895..10900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10915..10936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11053..11070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11224..11812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11304..11323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11983..12774,12775..12843))
                     /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     12606..12625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12844..12948)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12974..13429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13111..13132)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     13323..13342
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13556..13578
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13570..13587
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13571..13587
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13597..13616
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13597..13615
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   13645..13679
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gcatgggccc agatctccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
       61 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      121 atgggggcgg gggggggggg gggcgcgcgc crggcggggc ggggcggggc gaggggcggg
      181 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      241 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      301 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      361 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      421 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      481 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      541 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      601 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
      661 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
      721 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
      781 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
      841 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
      901 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
      961 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1021 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1081 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1141 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1201 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1261 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1321 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc gccaccatga
     1381 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     1441 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     1501 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     1561 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     1621 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     1681 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     1741 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     1801 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     1861 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     1921 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     1981 gagtttaaac catgcaagct tgatatcaag cttatcgata atcaacctct ggattacaaa
     2041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2401 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     2461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2581 cggatctccc tttgggccgc ctccccgcat cctgcaggta attctaccgg gtaggggagg
     2641 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     2701 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     2761 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     2821 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactaggc
     2881 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     2941 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     3001 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     3061 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     3121 gcctttcgac ctgcagccca agcttgtcga cgccaccatg gatgctttag acgattttga
     3181 cttagatatg cttggttcag acgcgttaga cgacttcgac ctagacatgt taggctcaga
     3241 tgcattggac gacttcgatt tagatatgtt gggctccgat gccctagatg actttgatct
     3301 agatatgcta ggtagtggag ggtcaggtgg tagtggcgga tcccagtatc tgcccgacac
     3361 agatgataga caccgaatcg aagagaaacg caagcgaacg tatgaaacct tcaaatcgat
     3421 catgaagaaa tcgcccttct cgggtccgac cgatcccagg cccccaccga gaaggattgc
     3481 ggtcccgtcc cgctcgtcgg ccagcgtgcc gaagcctgcg ccgcagccct accccttcac
     3541 gtcgagcctg agcacaatca attatgacga gttcccgacg atggtgttcc cctcgggaca
     3601 aatctcacaa gcctcggcgc tcgcaccagc gcctccccaa gtccttccgc aagcgcctgc
     3661 cccagcgcct gcaccggcaa tggtgtccgc cctcgcacag gcccctgcgc ccgtccccgt
     3721 gctcgcgcct ggaccgcccc aggcggtcgc tccaccggct ccgaagccga cgcaggccgg
     3781 agagggaaca ctctccgaag cacttcttca actccagttt gatgacgagg atcttggagc
     3841 actccttgga aactcgacag accctgcggt gtttaccgac ctcgcgtcag tagataactc
     3901 cgaatttcag cagcttttga accagggtat cccggtcgcg ccacatacaa cggagcccat
     3961 gttgatggaa taccccgaag caatcacgag acttgtgacg ggagcgcagc ggcctcccga
     4021 tcccgcaccc gcacctttgg gggcacctgg cctccctaac ggacttttga gcggcgacga
     4081 ggatttctcc tccatcgccg atatggattt ctcagccttg ctgtcacaga tttccagcgg
     4141 ctctggcagc ggcagccggg attccaggga agggatgttt ttgccgaagc ctgaggccgg
     4201 ctccgctatt agtgacgtgt ttgagggccg cgaggtgtgc cagccaaaac gaatccggcc
     4261 atttcatcct ccaggaagtc catgggccaa ccgcccactc cccgccagcc tcgcaccaac
     4321 accaaccggt ccagtacatg agccagtcgg gtcactgacc ccggcaccag tccctcagcc
     4381 actggatcca gcgcccgcag tgactcccga ggccagtcac ctgttggagg atcccgatga
     4441 agagacgagc caggctgtca aagcccttcg ggagatggcc gatactgtga ttccccagaa
     4501 ggaagaggct gcaatctgtg gccaaatgga cctttcccat ccgcccccaa ggggccatct
     4561 ggatgagctg acaaccacac ttgagtccat gaccgaggat ctgaacctgg actcacccct
     4621 gaccccggaa ttgaacgaga ttctggatac cttcctgaac gacgagtgcc tcttgcatgc
     4681 catgcatatc agcacaggac tgtccatctt cgacacatct ctgtttcata tgtcaagcgc
     4741 tgcaggtgct accatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     4801 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     4861 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     4921 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     4981 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     5041 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     5101 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     5161 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     5221 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     5281 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     5341 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     5401 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attcaagcgc
     5461 tgcaggtgcc accatggaca agaagtacag catcggcctg gccatcggca ccaactctgt
     5521 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     5581 caacaccgac cggcacagca tcaagaagaa cctgatcggc gccctgctgt tcgacagcgg
     5641 agaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     5701 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     5761 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     5821 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     5881 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     5941 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     6001 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     6061 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     6121 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     6181 cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     6241 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     6301 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     6361 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     6421 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     6481 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     6541 gaacggctac gccggctaca tcgatggcgg agccagccag gaagagttct acaagttcat
     6601 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     6661 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     6721 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     6781 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     6841 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     6901 gaacttcgag gaagtggtgg acaagggcgc cagcgcccag agcttcatcg agcggatgac
     6961 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     7021 gtacttcacc gtgtacaacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     7081 gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg gacctgctgt tcaagaccaa
     7141 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     7201 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     7261 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     7321 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     7381 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     7441 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     7501 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     7561 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     7621 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     7681 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     7741 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     7801 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     7861 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     7921 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     7981 cgatgtggac gctatcgtgc ctcagagctt tctgaaggac gactccatcg ataacaaagt
     8041 gctgactcgg agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     8101 gaagaagatg aagaactact ggcgccagct gctgaatgcc aagctgatta cccagaggaa
     8161 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     8221 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     8281 ctcccggatg aacactaagt acgacgagaa cgacaaactg atccgggaag tgaaagtgat
     8341 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     8401 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     8461 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     8521 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     8581 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     8641 gatccggaag cggcctctga tcgagacaaa cggcgaaaca ggcgagatcg tgtgggataa
     8701 gggccgggac tttgccaccg tgcggaaagt gctgtctatg ccccaagtga atatcgtgaa
     8761 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     8821 cgacaagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     8881 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     8941 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     9001 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     9061 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     9121 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     9181 gtacctggcc agccactatg agaagctgaa ggggggcagt ggcgggagcg ggcgcgccta
     9241 tccctacgat gtgcccgatt acgctggtgg atccggcggg tcagatgaga tggaagagtg
     9301 ctctcagcac ggcggatctg gaggttctac aggctgcgtg gtcatagtgg gcaggatcgt
     9361 cttgtccgga tccggcacta gtgcgcccat cacggcgtac gcccagcaga cgagaggcct
     9421 cctagggtgt ataatcacca gcctgactgg ccgggacaaa aaccaagtgg agggtgaggt
     9481 ccagatcgtg tcaactgcta cccaaacctt gctggcaacg tgcatcaatg gggtatgctg
     9541 gacagtctac cacggggccg gaacgaggac catcgcatca cccaagggtc ctgtcatcca
     9601 gatgtatacc aatgtggaca aagaccttgt gggctggccc gctcctcaag gttcccgctc
     9661 attgacaccc tgtacctgcg gctcctcgga cctttacctg gtcacgaggc acgccgatgt
     9721 cattcccgtg cgccggcgag gtgatcgcag gggtagcctg ctttcgcccc ggcccatttc
     9781 ctacttgaaa ggctcctcgg ggggtccgct gttgtgcccc gcgggacacg ccgtgggcct
     9841 attcagggcc gcggtgtgca cccgtggagt ggctaaagcg gtgtacttta tccctgtgga
     9901 gaacctagag acaaccatga gatccccggt gttcacggac aactcctctc caccagcagt
     9961 caccctgacg cacgggggtt caggcggtag tggctccccc gaggataatg agcagaaaca
    10021 gctgtttgtg gaacagcaca aacactacct ggacgagatc atcgagcaga tcagcgagtt
    10081 ctccaagaga gtgatcctgg ccgacgctaa tctggacaag gtgctgagcg cctacaacaa
    10141 gcacagagac aagcctatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
    10201 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
    10261 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
    10321 cgagacacgg atcgacctgt ctcagctggg aggcgacgcc tatccctatg acgtgcccga
    10381 ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag atcctaagaa
    10441 aaagcggaaa gtggacggca ttggtagtgg gagcaacgga tgactcgagc tagttaaaag
    10501 ttttgttact ttatagaaga aattttgagt ttttgttttt ttttaataaa taaataaaca
    10561 taaataaatt gtttgttgaa tttattatta gtatgtaagt gtaaatataa taaaacttaa
    10621 tatctattca aattaataaa taaacctcga tatacagacc gataaaacac atgcgtcaat
    10681 tttacgcatg attatcttta acgtacgtca caatatgatt atctttctag ggttaatcta
    10741 gtatacgcgt atgcggccgc ttaattaatc cagcttttgt tccctttagt gagggttaat
    10801 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    10861 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    10921 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    10981 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    11041 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    11101 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    11161 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    11221 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    11281 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    11341 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    11401 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    11461 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    11521 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    11581 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    11641 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
    11701 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    11761 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    11821 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    11881 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    11941 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    12001 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    12061 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    12121 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    12181 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    12241 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    12301 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    12361 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    12421 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    12481 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    12541 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    12601 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    12661 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    12721 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    12781 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    12841 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    12901 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    12961 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
    13021 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
    13081 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
    13141 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
    13201 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
    13261 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
    13321 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
    13381 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
    13441 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
    13501 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
    13561 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
    13621 ttggggcgcg ccattctaga ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc
    13681 attcttgaaa tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga
    13741 catctcagtc gccgcttgga gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact
    13801 gattttgaac tataacgacc gcgtgagtca aaatgacgca tgattatctt ttacgtgact
    13861 tttaagattt aactcatacg ataattatat tgttatttca tgttctactt acgtgataac
    13921 ttattatata tatattttct tgttatagat atcaactaga atgcta
//
