LOCUS       sequence-337581-       14400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Vp64-dCas9 activation system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14400)
  AUTHORS   Penev A, Bazley A, Shen M, Boeke JD, Savage SA, Sfeir A
  TITLE     Alternative splicing is a developmental switch for hTERT expression.
  JOURNAL   Mol Cell. 2021 Apr 6. pii: S1097-2765(21)00228-8. doi: 
            10.1016/j.molcel.2021.03.033.
  PUBMED    33852895
REFERENCE   2  (bases 1 to 14400)
  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: "Mol Cell. 
            2021 Apr 6. pii: S1097-2765(21)00228-8. doi: 
            10.1016/j.molcel.2021.03.033."
COMMENT     Sequence Label: pLenti Vp64-dCas9 T2A mTagBFP2
FEATURES             Location/Qualifiers
     source          1..14400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..55)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        40..369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      220..355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     282..301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        417..464
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             483..857
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    987..1108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1024..1043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1078..1097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1157..1173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1157..1173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1170..1192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1181..1197
                     /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(1205..1211,1212..1229,1230..1235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1250..1271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1388..1405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1559..2147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1639..1658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2318..3109,3110..3178))
                     /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     2941..2960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3179..3283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3358..3377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3549..3928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3930..4128
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4083..4103
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4146..4326
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4373..4498
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4991..5224
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5409..5453
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5751..5868
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5998..7176
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6229..7167
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7124..7144
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7192..7239
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(7240..7263,7264..7266,7267..9822,9823..9825,
                     9826..11340)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11395..11415
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11440..11589
                     /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             11611..11664
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             11683..12378
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="ELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVE
                     GGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQ
                     DTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLV
                     GGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDL
                     PSKLGHKLN"
     misc_feature    12403..12991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12456..12476)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12874..12885)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12994..13010)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12995..13011)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             13516..13696
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(13722..13739)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13728..13952
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(13998..14400,1..26)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14085..14104)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14295..14316
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
       61 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
      241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      301 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      361 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
      421 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
      481 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
      541 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
      601 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
      661 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
      721 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
      781 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
      841 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
      901 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
      961 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     1021 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1081 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     1141 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1201 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1261 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1321 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1381 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1441 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1501 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1561 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1621 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1681 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1741 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1801 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1861 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1921 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1981 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2041 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2101 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2161 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2221 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2281 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2341 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2401 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2461 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2521 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2581 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2641 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2701 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2761 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2821 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2881 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2941 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3001 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3061 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3121 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3181 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3241 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3301 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     3361 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     3421 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     3481 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     3541 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     3601 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3661 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3721 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3781 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3841 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3901 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     3961 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     4021 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     4081 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct
     4141 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     4201 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4261 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4321 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     4381 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     4441 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     4501 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     4561 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     4621 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     4681 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     4741 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     4801 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     4861 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     4921 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     4981 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     5041 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     5101 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     5161 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     5221 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     5281 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     5341 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     5401 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     5461 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     5521 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     5581 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     5641 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     5701 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     5761 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     5821 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     5881 agggacagca gagatccagt ttggttaatt agctagctgc aaagatggat aaagttttaa
     5941 acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc
     6001 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     6061 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     6121 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     6181 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     6241 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     6301 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     6361 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     6421 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     6481 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     6541 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     6601 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     6661 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     6721 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     6781 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     6841 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     6901 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     6961 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     7021 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     7081 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     7141 gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat gaaaaggccg
     7201 gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg acaagaagta cagcatcggc
     7261 ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     7321 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     7381 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     7441 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     7501 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     7561 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     7621 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     7681 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     7741 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     7801 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     7861 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     7921 ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg
     7981 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     8041 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     8101 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     8161 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     8221 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     8281 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     8341 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     8401 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     8461 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     8521 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     8581 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     8641 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     8701 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     8761 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     8821 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     8881 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     8941 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     9001 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     9061 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     9121 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     9181 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     9241 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     9301 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     9361 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     9421 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     9481 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     9541 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     9601 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     9661 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     9721 gacatcaacc ggctgtccga ctacgatgtg gaccacatcg tgcctcagag ctttctgaag
     9781 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca aggcccgggg caagagcgac
     9841 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     9901 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     9961 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
    10021 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
    10081 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
    10141 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
    10201 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
    10261 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
    10321 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
    10381 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
    10441 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
    10501 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
    10561 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
    10621 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
    10681 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
    10741 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
    10801 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
    10861 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
    10921 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
    10981 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
    11041 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
    11101 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
    11161 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
    11221 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
    11281 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
    11341 agcgctggag gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag
    11401 aaaaagagga aggtggcggc cgctggatcc ggacgggctg acgcattgga cgattttgat
    11461 ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat
    11521 gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg
    11581 gacatgctga ttaactgtac cggcagtgga gagggcagag gaagtctgct aacatgcggt
    11641 gacgtcgagg agaatcctgg cccaatggtg agcaagggtg aggagctgat taaggagaac
    11701 atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc
    11761 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
    11821 ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc
    11881 ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc
    11941 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
    12001 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc
    12061 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac
    12121 cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc
    12181 catctgatcg caaacgccaa gaccacatat agatccaaga aacccgctaa gaacctcaag
    12241 atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag
    12301 acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg
    12361 gggcacaagc ttaattgaga attcgatatc aagcttatcg gtaatcaacc tctggattac
    12421 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
    12481 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
    12541 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
    12601 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
    12661 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
    12721 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
    12781 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
    12841 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
    12901 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
    12961 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg agacctagaa
    13021 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
    13081 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
    13141 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
    13201 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
    13261 ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc
    13321 tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa
    13381 ggagagaaca cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
    13441 gaagtattag agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg
    13501 catccggact gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
    13561 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
    13621 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
    13681 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
    13741 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
    13801 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
    13861 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
    13921 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
    13981 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
    14041 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
    14101 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
    14161 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
    14221 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
    14281 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
    14341 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
//
