LOCUS       sequence-235074-       12435 bp    DNA     circular SYN 14-APR-2025
DEFINITION  encodes a GAG-dCAS9-VPR fusion for targeted transcriptional 
            activation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12435)
  AUTHORS   Mangeot PE, Risson V, Fusil F, Marnef A, Laurent E, Blin J, 
            Mournetas V, Massourides E, Sohier TJM, Corbin A, Aube F, Teixeira 
            M, Pinset C, Schaeffer L, Legube G, Cosset FL, Verhoeyen E, Ohlmann 
            T, Ricci EP
  TITLE     Genome editing in primary cells and in vivo using viral-derived 
            Nanoblades loaded with Cas9-sgRNA ribonucleoproteins.
  JOURNAL   Nat Commun. 2019 Jan 3;10(1):45. doi: 10.1038/s41467-018-07845-z.
  PUBMED    30604748
REFERENCE   2  (bases 1 to 12435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-01-3"; volume: "10(1):45. doi"; pages: " 
            10.1038/s41467-018-07845-z"
COMMENT     Sequence Label: BICstim-Gag-dCAS9-VPR
FEATURES             Location/Qualifiers
     source          1..12435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        431..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          746..1318
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        3057..3095
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3204..3213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3210..3236,3237..3239,3240..5723,5724..5729,5730..5795,
                     5796..5798,5799..7313)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKNVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7326..7346
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    7383..7407
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7437..7586
                     /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             7611..7631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8061..8417
                     /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
                     EDFSSIADMDFSALL"
     CDS             8436..9005
                     /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"
     primer_bind     complement(9094..9113)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9159..9214
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9213..9232)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(9572..9588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9572..9588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9585..9607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9596..9612
                     /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(9620..9626,9627..9644,9645..9650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9665..9686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9803..9820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9974..10562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10054..10073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10733..11524,11525..11593))
                     /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     11356..11375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11594..11698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11724..12179)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11861..11882)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12073..12092
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12306..12328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12320..12337
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12321..12337
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12347..12366
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12347..12365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt
       61 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      661 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      721 gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt
      781 cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat
      841 gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct
      901 actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt
      961 aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg
     1021 taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc
     1081 agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat
     1141 aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc tggtcatcat
     1201 cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc
     1261 tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt tcctacagct
     1321 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tcctcgaggg
     1381 ccggatccga attcctgtat ttgtctgaaa acatgggcca ggctgttacc acccccttaa
     1441 gtttgacttt agaccactgg aaggatgtcg aacggacagc ccacaacctg tcggtagagg
     1501 ttagaaaaag gcgctgggtt acattctgct ctgcagaatg gccaaccttc aacgtcggat
     1561 ggccacgaga cggcactttt aacccagaca ttattacaca ggttaagatc aaggtcttct
     1621 cacctggccc acatggacat ccggatcagg tcccctacat cgtgacctgg gaagctatag
     1681 cagtagaccc ccctccctgg gtcagaccct tcgtgcaccc taaacctccc ctctctcttc
     1741 ccccttcagc cccctctctc ccacctgaac ccccactctc gaccccgccc cagtcctccc
     1801 tctatccggc tctcacttct cctttaaaca ccaaacctag gcctcaagtc cttcctgata
     1861 gcggaggacc actcattgat ctactcacgg aggaccctcc gccttaccgg gacccagggc
     1921 caccctctcc tgacgggaac ggcgatagcg gagaagtggc ccctacagaa ggagcccctg
     1981 acccttcccc aatggtatcc cgcctgcggg gaagaaaaga accccccgtg gcggattcta
     2041 ctacctctca ggcgttcccc cttcgcctgg gagggaatgg acagtatcaa tactggccat
     2101 tttcctcctc tgacctctat aactggaaaa ataacaaccc ctctttctcc gaggacccag
     2161 ctaaattgac agctttgatc gagtccgttc tccttactca tcagcccact tgggatgact
     2221 gccaacagct attagggacc ctgctgacgg gagaagaaaa acagcgagtg ctcctagagg
     2281 cccgaaaggc ggttcgaggg gaggacggac gcccaactca gctgcccaat gacattaatg
     2341 atgcttttcc cttggaacgt cccgactggg actacaacac ccaacgaggt aggaaccacc
     2401 tagtccacta tcgccagttg ctcctagcgg gtctccaaaa cgcgggcaga agccccacca
     2461 atttggccaa ggtaaaaggg ataacccagg gacctaatga gtctccctca gcctttttag
     2521 agagactcaa ggaggcctat cgcagataca ctccttatga ccctgaggac ccagggcaag
     2581 aaaccaatgt ggccatgtca ttcatctggc agtccgcccc ggatatcggg cgaaagttag
     2641 agcggttaga agatttgaag agtaagacct taggagactt agtgagggaa gctgaaaaga
     2701 tctttaataa acgagaaacc ccggaagaaa gagaggaacg tattaggaga gaaacagagg
     2761 aaaaggaaga acgccgtagg gcagaggatg tgcagagaga gaaggagagg gaccgcagaa
     2821 gacatagaga aatgagtaag ttgctggcta ctgtcgttag cgggcagaga caggatagac
     2881 agggaggaga gcgaaggagg ccccaactcg accacgacca gtgtgcctac tgcaaagaaa
     2941 agggacattg ggctagagat tgccccaaga agccaagagg accccgggga ccacgacccc
     3001 aggcctccct cctgacgcgt agttccctgt atccagccct cacacctacc ggtggcggta
     3061 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     3121 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     3181 ggactctaga ggatcgaacc cttgccacca tggacaagaa gtactccatt gggctcgcta
     3241 tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa
     3301 aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc
     3361 tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca
     3421 gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg
     3481 ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata
     3541 aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa
     3601 agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact
     3661 tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg
     3721 agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga
     3781 cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa
     3841 tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg
     3901 gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca
     3961 actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct
     4021 acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt
     4081 tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg
     4141 agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag
     4201 acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt
     4261 tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg
     4321 aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg
     4381 taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc
     4441 cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc
     4501 cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact
     4561 atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag
     4621 agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct
     4681 tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac
     4741 actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca
     4801 cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc
     4861 tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa
     4921 agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc
     4981 tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg
     5041 agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga
     5101 tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc
     5161 tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc
     5221 gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc
     5281 ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag
     5341 cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc
     5401 cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa
     5461 tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc
     5521 agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac
     5581 tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc
     5641 tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca
     5701 atcggctctc cgactacgac gtggctgcta tcgtgcccca gtcttttctc aaagatgatt
     5761 ctattgataa taaagtgttg acaagatccg ataaagctag agggaagagt gataacgtcc
     5821 cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac
     5881 tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt
     5941 tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg
     6001 tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc
     6061 gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc
     6121 agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg
     6181 cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg
     6241 gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca
     6301 aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta
     6361 cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag
     6421 aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc
     6481 aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc
     6541 tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat
     6601 acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga
     6661 aagggaagtc taaaaaactc aaaaacgtca aggaactgct gggcatcaca atcatggagc
     6721 gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca
     6781 aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga
     6841 aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta
     6901 aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag
     6961 ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg
     7021 agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc
     7081 tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc
     7141 acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca
     7201 tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt
     7261 caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg
     7321 ctgaccccaa gaagaagagg aaggtgtcgc cagggatccg tcgacttgac gcgttgatat
     7381 caacaagttt gtacaaaaaa gcaggctaca aagaggccag cggttccgga cgggctgacg
     7441 cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat tttgaccttg
     7501 acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc agtgacgccc
     7561 ttgatgattt cgacctggac atgctgatta actctagaag ttccggatct ccgaaaaaga
     7621 aacgcaaagt tggtagccag tacctgcccg acaccgacga ccggcaccgg atcgaggaaa
     7681 agcggaagcg gacctacgag acattcaaga gcatcatgaa gaagtccccc ttcagcggcc
     7741 ccaccgaccc tagacctcca cctagaagaa tcgccgtgcc cagcagatcc agcgccagcg
     7801 tgccaaaacc tgccccccag ccttacccct tcaccagcag cctgagcacc atcaactacg
     7861 acgagttccc taccatggtg ttccccagcg gccagatctc tcaggcctct gctctggctc
     7921 cagcccctcc tcaggtgctg cctcaggctc ctgctcctgc accagctcca gccatggtgt
     7981 ctgcactggc tcaggcacca gcacccgtgc ctgtgctggc tcctggacct ccacaggctg
     8041 tggctccacc agcccctaaa cctacacagg ccggcgaggg cacactgtct gaagctctgc
     8101 tgcagctgca gttcgacgac gaggatctgg gagccctgct gggaaacagc accgatcctg
     8161 ccgtgttcac cgacctggcc agcgtggaca acagcgagtt ccagcagctg ctgaaccagg
     8221 gcatccctgt ggcccctcac accaccgagc ccatgctgat ggaatacccc gaggccatca
     8281 cccggctcgt gacaggcgct cagaggcctc ctgatccagc tcctgcccct ctgggagcac
     8341 caggcctgcc taatggactg ctgtctggcg acgaggactt cagctctatc gccgatatgg
     8401 atttctcagc cttgctgggc tctggcagcg gcagccggga ttccagggaa gggatgtttt
     8461 tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt tgagggccgc gaggtgtgcc
     8521 agccaaaacg aatccggcca tttcatcctc caggaagtcc atgggccaac cgcccactcc
     8581 ccgccagcct cgcaccaaca ccaactggtc cagtacatga gccagtcggg tcactgaccc
     8641 cggcaccagt ccctcagcca ctggatccag cgcccgcagt gactcccgag gccagtcacc
     8701 tgttggagga tcccgatgaa gagacgagcc aggctgtcaa agcccttcgg gagatggccg
     8761 atactgtgat tccccagaag gaagaggctg caatctgtgg ccaaatggac ctttcccatc
     8821 cgcccccaag gggccatctg gatgagctga caaccacact tgagtccatg accgaggatc
     8881 tgaacctgga ctcacccctg accccggaat tgaacgagat tctggatacc ttcctgaacg
     8941 acgagtgcct cttgcatgcc atgcatatca gcacaggact gtccatcttc gacacatctc
     9001 tgttttgagg tacctgcatc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     9061 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     9121 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     9181 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     9241 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     9301 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     9361 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     9421 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     9481 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     9541 cctcgacgga tcggccgcaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt
     9601 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     9661 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     9721 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     9781 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     9841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     9901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     9961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    10021 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    10081 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    10141 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    10201 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    10261 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    10321 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    10381 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    10441 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    10501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    10561 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    10621 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    10681 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    10741 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    10801 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    10861 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    10921 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    10981 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    11041 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    11101 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    11161 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    11221 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    11281 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    11341 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    11401 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    11461 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    11521 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    11581 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    11641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    11701 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
    11761 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
    11821 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
    11881 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
    11941 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
    12001 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
    12061 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
    12121 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
    12181 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
    12241 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
    12301 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
    12361 atagggcgaa ttggagctcc accgcggtgg cggccgctct agagagcttg gcccattgca
    12421 tacgttgtat ccata
//
