LOCUS       sequence-397645-        9906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. Pyogenes dCas9 fused to LargeBiT fragment of split 
            NanoLuc luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9906)
  AUTHORS   van der Veer HJ, van Aalen EA, Michielsen CMS, Hanckmann ETL, 
            Deckers J, van Borren MMGJ, Flipse J, Loonen AJM, Schoeber JPH, 
            Merkx M
  TITLE     Glow-in-the-Dark Infectious Disease Diagnostics Using 
            CRISPR-Cas9-Based Split Luciferase Complementation.
  JOURNAL   ACS Cent Sci. 2023 Mar 15;9(4):657-667. doi: 
            10.1021/acscentsci.2c01467. eCollection 2023 Apr 26.
  PUBMED    37122471
REFERENCE   2  (bases 1 to 9906)
  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: 
            "10.1021/acscentsci.2c01467"; journalName: "ACS Cent Sci"; date: 
            "2023-03-15- 15"; volume: "9"; issue: "4"; pages: "657-667"
COMMENT     Sequence Label: pET28a(+)-SpdCas9-LBiT_Strep-Tag-II
FEATURES             Location/Qualifiers
     source          1..9906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(167..186)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     377..398
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(627..1442)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(627..1442)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1350..1369
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1434..1453
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1564..2152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2053..2072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2306..2323
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2338..2477
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2403..2422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2563..2585
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2579..2770)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3579..4661)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4623..4642
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4662..4739)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4945..4964
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5048..5067
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5048..5066
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5067..5091
                     /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)."
     RBS             5106..5128
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(5135..5161,5162..5164,5165..7651,7652..7654,
                     7655..9238)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9785..9808
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             9818..9835
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9888..9906)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(9902..9906,1..43)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
       61 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      121 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      181 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      241 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      301 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      361 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      421 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      481 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
      541 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      601 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
      661 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
      721 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
      781 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
      841 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
      901 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
      961 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     1021 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     1081 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     1141 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     1201 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     1261 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     1321 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     1381 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     1441 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
     1501 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1561 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1621 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1681 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1741 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1801 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1861 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1921 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1981 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2041 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2101 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2161 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2221 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2281 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2341 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     2401 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     2461 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2521 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2581 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     2641 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     2701 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     2761 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     2821 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     2881 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     2941 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     3001 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     3061 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     3121 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     3181 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     3241 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     3301 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     3361 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     3421 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     3481 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     3541 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3601 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3661 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     3721 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     3781 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     3841 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     3901 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     3961 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     4021 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     4081 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     4141 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     4201 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     4261 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     4321 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     4381 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     4441 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     4501 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     4561 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     4621 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     4681 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     4741 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     4801 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     4861 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     4921 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     4981 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     5041 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     5101 ataattttgt ttaactttaa gaaggagata taccatggat aagaaatact caataggctt
     5161 agctatcggc acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc
     5221 taaaaagttc aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg
     5281 ggctctttta tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg
     5341 tagaaggtat acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga
     5401 gatggcgaaa gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga
     5461 agacaagaag catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca
     5521 tgagaaatat ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc
     5581 ggatttgcgc ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt
     5641 gattgaggga gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt
     5701 acaaacctac aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa
     5761 agcgattctt tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct
     5821 ccccggtgag aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac
     5881 ccctaatttt aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga
     5941 tacttacgat gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt
     6001 gtttttggca gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa
     6061 tactgaaata actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca
     6121 tcaagacttg actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga
     6181 aatctttttt gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca
     6241 agaagaattt tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt
     6301 attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc
     6361 tattccccat caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt
     6421 ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc
     6481 ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc
     6541 tgaagaaaca attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca
     6601 atcatttatt gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc
     6661 aaaacatagt ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata
     6721 tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt
     6781 tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt
     6841 caaaaaaata gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc
     6901 ttcattaggt acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa
     6961 tgaagaaaat gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag
     7021 ggagatgatt gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa
     7081 acagcttaaa cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg
     7141 tattagggat aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc
     7201 caatcgcaat tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca
     7261 aaaagcacaa gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg
     7321 tagccctgct attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa
     7381 agtaatgggg cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac
     7441 aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa
     7501 agaattagga agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga
     7561 aaagctctat ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga
     7621 tattaatcgt ttaagtgatt atgatgtcga tgccattgtt ccacaaagtt tccttaaaga
     7681 cgattcaata gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa
     7741 cgttccaagt gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc
     7801 caagttaatc actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag
     7861 tgaacttgat aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa
     7921 gcatgtggca caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact
     7981 tattcgagag gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga
     8041 tttccaattc tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct
     8101 aaatgccgtc gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt
     8161 ctatggtgat tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat
     8221 aggcaaagca accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga
     8281 aattacactt gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac
     8341 tggagaaatt gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat
     8401 gccccaagtc aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc
     8461 aattttacca aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa
     8521 aaaatatggt ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt
     8581 ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat
     8641 ggaaagaagt tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga
     8701 agttaaaaaa gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg
     8761 tcgtaaacgg atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc
     8821 aagcaaatat gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc
     8881 agaagataac gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat
     8941 tattgagcaa atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa
     9001 agttcttagt gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat
     9061 tattcattta tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac
     9121 aacaattgat cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca
     9181 tcaatccatc actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgacac
     9241 cggtgggggt agcggcggct cggggggtag tggtggaagc gggggttcaa agcttactag
     9301 tgtcttcaca ctcgaagatt tcgttgggga ctgggaacag acagccgcct acaacctgga
     9361 ccaagtcctt gaacagggag gtgtgtccag tttgctgcag aatctcgccg tgtccgtaac
     9421 tccgatccaa aggattgtcc ggagcggtga aaatgccctg aagatcgaca tccatgtcat
     9481 catcccgtat gaaggtctga gcgccgacca aatggcccag atcgaagagg tgtttaaggt
     9541 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg ccctatggca cactggtaat
     9601 cgacggggtt acgccgaaca tgctgaacta tttcggacgg ccgtatgaag gcatcgccgt
     9661 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     9721 cgagcgcctg atcacccccg acggctccat gctgttccga gtaaccatca acagcggtgg
     9781 aagctggagc catccgcagt ttgaaaaata actcgagcac caccaccacc accactgaga
     9841 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     9901 actagc
//
