LOCUS       sequence-229834-       12436 bp    DNA     circular SYN 15-DEC-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 12436)
  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 12436)
  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 
            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..12436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          792..1365
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1373..1392
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        3104..3142
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3251..3260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3257..3283,3284..3286,3287..5770,5771..5776,5777..5842,
                     5843..5845,5846..7360)
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7373..7393
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    7430..7454
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7484..7633
                     /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             7658..7678
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8108..8464
                     /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             8483..9052
                     /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(9140..9159)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9205..9260
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9259..9278)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(9618..9634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9618..9634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9631..9653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9642..9658
                     /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(9666..9672,9673..9690,9691..9696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9711..9732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9849..9866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10020..10608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10100..10119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11077..11456)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     11403..11422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11641..11745)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11771..12226)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11908..11929)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12120..12139
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12353..12375
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12367..12384
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12368..12384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12394..12413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12394..12412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      721 gctgttttga cctccataga agacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt cttgtaactt tttcgttaaa ctttagcttg
     1021 catttgtaac gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc
     1081 taatcacttt tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag
     1141 agaacaattg ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg
     1201 cgtggaaata ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct
     1261 ttatggttac aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg
     1321 ggcccctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1381 ctggttattg tgctgtctca tcattttggc aaagaattcc tcgagggccg gatccgaatt
     1441 cctgtatttg tctgaaaaca tgggccaggc tgttaccacc cccttaagtt tgactttaga
     1501 ccactggaag gatgtcgaac ggacagccca caacctgtcg gtagaggtta gaaaaaggcg
     1561 ctgggttaca ttctgctctg cagaatggcc aaccttcaac gtcggatggc cacgagacgg
     1621 cacttttaac ccagacatta ttacacaggt taagatcaag gtcttctcac ctggcccaca
     1681 tggacatccg gatcaggtcc cctacatcgt gacctgggaa gctatagcag tagacccccc
     1741 tccctgggtc agacccttcg tgcaccctaa acctcccctc tctcttcccc cttcagcccc
     1801 ctctctccca cctgaacccc cactctcgac cccgccccag tcctccctct atccggctct
     1861 cacttctcct ttaaacacca aacctaggcc tcaagtcctt cctgatagcg gaggaccact
     1921 cattgatcta ctcacggagg accctccgcc ttaccgggac ccagggccac cctctcctga
     1981 cgggaacggc gatagcggag aagtggcccc tacagaagga gcccctgacc cttccccaat
     2041 ggtatcccgc ctgcggggaa gaaaagaacc ccccgtggcg gattctacta cctctcaggc
     2101 gttccccctt cgcctgggag ggaatggaca gtatcaatac tggccatttt cctcctctga
     2161 cctctataac tggaaaaata acaacccctc tttctccgag gacccagcta aattgacagc
     2221 tttgatcgag tccgttctcc ttactcatca gcccacttgg gatgactgcc aacagctatt
     2281 agggaccctg ctgacgggag aagaaaaaca gcgagtgctc ctagaggccc gaaaggcggt
     2341 tcgaggggag gacggacgcc caactcagct gcccaatgac attaatgatg cttttccctt
     2401 ggaacgtccc gactgggact acaacaccca acgaggtagg aaccacctag tccactatcg
     2461 ccagttgctc ctagcgggtc tccaaaacgc gggcagaagc cccaccaatt tggccaaggt
     2521 aaaagggata acccagggac ctaatgagtc tccctcagcc tttttagaga gactcaagga
     2581 ggcctatcgc agatacactc cttatgaccc tgaggaccca gggcaagaaa ccaatgtggc
     2641 catgtcattc atctggcagt ccgccccgga tatcgggcga aagttagagc ggttagaaga
     2701 tttgaagagt aagaccttag gagacttagt gagggaagct gaaaagatct ttaataaacg
     2761 agaaaccccg gaagaaagag aggaacgtat taggagagaa acagaggaaa aggaagaacg
     2821 ccgtagggca gaggatgtgc agagagagaa ggagagggac cgcagaagac atagagaaat
     2881 gagtaagttg ctggctactg tcgttagcgg gcagagacag gatagacagg gaggagagcg
     2941 aaggaggccc caactcgacc acgaccagtg tgcctactgc aaagaaaagg gacattgggc
     3001 tagagattgc cccaagaagc caagaggacc ccggggacca cgaccccagg cctccctcct
     3061 gacgcgtagt tccctgtatc cagccctcac acctaccggt ggcggtaggc gtgtacggtg
     3121 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
     3181 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga ctctagagga
     3241 tcgaaccctt gccaccatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag
     3301 cgtcggctgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct
     3361 gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc
     3421 cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag
     3481 aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga
     3541 ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg
     3601 ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat
     3661 atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta
     3721 tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa
     3781 cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct
     3841 tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag
     3901 gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa
     3961 cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa
     4021 cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct
     4081 cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa
     4141 cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc
     4201 tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct
     4261 gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc
     4321 taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt
     4381 tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag
     4441 agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca
     4501 cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga
     4561 taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg taggccccct
     4621 cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc
     4681 ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat
     4741 gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta
     4801 cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag
     4861 aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac
     4921 gaaccggaaa gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt
     4981 cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca
     5041 cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat
     5101 tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg
     5161 cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg
     5221 atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag
     5281 tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca
     5341 gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg
     5401 ccagggggac agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa
     5461 gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa
     5521 gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa
     5581 gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat
     5641 ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta
     5701 cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga
     5761 ctacgacgtg gctgctatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa
     5821 agtgttgaca agatccgata aagctagagg gaagagtgat aacgtcccct cagaagaagt
     5881 tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg
     5941 gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg
     6001 cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct
     6061 cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt
     6121 tattactctg aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt
     6181 gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac
     6241 tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt
     6301 gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa
     6361 gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg
     6421 agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga
     6481 caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt
     6541 taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa
     6601 cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga
     6661 ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa
     6721 aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga
     6781 aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat
     6841 cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc
     6901 tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt
     6961 cttgtatctg gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa
     7021 gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga
     7081 attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa
     7141 taagcacagg gataagccca tcagggagca ggcagaaaac attatccact tgtttactct
     7201 gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg
     7261 gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct
     7321 ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa
     7381 gaagaggaag gtgtcgccag ggatccgtcg acttgacgcg ttgatatcaa caagtttgta
     7441 caaaaaagca ggctacaaag aggccagcgg ttccggacgg gctgacgcat tggacgattt
     7501 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc
     7561 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga
     7621 cctggacatg ctgattaact ctagaagttc cggatctccg aaaaagaaac gcaaagttgg
     7681 tagccagtac ctgcccgaca ccgacgaccg gcaccggatc gaggaaaagc ggaagcggac
     7741 ctacgagaca ttcaagagca tcatgaagaa gtcccccttc agcggcccca ccgaccctag
     7801 acctccacct agaagaatcg ccgtgcccag cagatccagc gccagcgtgc caaaacctgc
     7861 cccccagcct taccccttca ccagcagcct gagcaccatc aactacgacg agttccctac
     7921 catggtgttc cccagcggcc agatctctca ggcctctgct ctggctccag cccctcctca
     7981 ggtgctgcct caggctcctg ctcctgcacc agctccagcc atggtgtctg cactggctca
     8041 ggcaccagca cccgtgcctg tgctggctcc tggacctcca caggctgtgg ctccaccagc
     8101 ccctaaacct acacaggccg gcgagggcac actgtctgaa gctctgctgc agctgcagtt
     8161 cgacgacgag gatctgggag ccctgctggg aaacagcacc gatcctgccg tgttcaccga
     8221 cctggccagc gtggacaaca gcgagttcca gcagctgctg aaccagggca tccctgtggc
     8281 ccctcacacc accgagccca tgctgatgga ataccccgag gccatcaccc ggctcgtgac
     8341 aggcgctcag aggcctcctg atccagctcc tgcccctctg ggagcaccag gcctgcctaa
     8401 tggactgctg tctggcgacg aggacttcag ctctatcgcc gatatggatt tctcagcctt
     8461 gctgggctct ggcagcggca gccgggattc cagggaaggg atgtttttgc cgaagcctga
     8521 ggccggctcc gctattagtg acgtgtttga gggccgcgag gtgtgccagc caaaacgaat
     8581 ccggccattt catcctccag gaagtccatg ggccaaccgc ccactccccg ccagcctcgc
     8641 accaacacca actggtccag tacatgagcc agtcgggtca ctgaccccgg caccagtccc
     8701 tcagccactg gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc
     8761 cgatgaagag acgagccagg ctgtcaaagc ccttcgggag atggccgata ctgtgattcc
     8821 ccagaaggaa gaggctgcaa tctgtggcca aatggacctt tcccatccgc ccccaagggg
     8881 ccatctggat gagctgacaa ccacacttga gtccatgacc gaggatctga acctggactc
     8941 acccctgacc ccggaattga acgagattct ggataccttc ctgaacgacg agtgcctctt
     9001 gcatgccatg catatcagca caggactgtc catcttcgac acatctctgt tttgaggtac
     9061 ctgcagctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     9121 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     9181 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     9241 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     9301 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     9361 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     9421 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     9481 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     9541 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacggatcgg
     9601 ccgcaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     9661 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     9721 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     9781 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     9841 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     9901 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     9961 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
    10021 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
    10081 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
    10141 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
    10201 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
    10261 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
    10321 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
    10381 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
    10441 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
    10501 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
    10561 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
    10621 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
    10681 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
    10741 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
    10801 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
    10861 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
    10921 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
    10981 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
    11041 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
    11101 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
    11161 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg gttagctcct tcggtcctcc
    11221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    11281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    11341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    11401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    11461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    11521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    11581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    11641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    11701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    11761 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
    11821 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
    11881 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
    11941 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
    12001 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
    12061 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
    12121 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
    12181 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
    12241 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
    12301 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
    12361 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
    12421 gagctccacc gcggtg
//
