LOCUS       pET28a(+)-SpdCas        9991 bp DNA     circular SYN 21-FEB-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9991)
  AUTHORS   iGEM Eindhoven 2019
  TITLE     Direct Submission
  JOURNAL   Exported Feb 21, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(560..1375)
                     /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"
     rep_origin      1497..2085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2271..2413
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2515..2706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3515..4597)
                     /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"
     promoter        complement(4598..4675)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /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             5042..5064
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5071..9174
                     /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"
     misc_feature    9238..9270
                     /label=SmallBiT 2.5 µM NanoBiT
                     /label=SBitNL 2.5uM
     CDS             9286..9309
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    9319..9792
                     /label=LargeBiT NanoBiT
                     /label=LBitNL
     CDS             9802..9825
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             9835..9852
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      9919..9966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatatacc atggataaga aatactcaat aggcttagct
     5101 atcggcacaa atagcgtcgg atgggcggtg atcactgatg aatataaggt tccgtctaaa
     5161 aagttcaagg ttctgggaaa tacagaccgc cacagtatca aaaaaaatct tataggggct
     5221 cttttatttg acagtggaga gacagcggaa gcgactcgtc tcaaacggac agctcgtaga
     5281 aggtatacac gtcggaagaa tcgtatttgt tatctacagg agattttttc aaatgagatg
     5341 gcgaaagtag atgatagttt ctttcatcga cttgaagagt cttttttggt ggaagaagac
     5401 aagaagcatg aacgtcatcc tatttttgga aatatagtag atgaagttgc ttatcatgag
     5461 aaatatccaa ctatctatca tctgcgaaaa aaattggtag attctactga taaagcggat
     5521 ttgcgcttaa tctatttggc cttagcgcat atgattaagt ttcgtggtca ttttttgatt
     5581 gagggagatt taaatcctga taatagtgat gtggacaaac tatttatcca gttggtacaa
     5641 acctacaatc aattatttga agaaaaccct attaacgcaa gtggagtaga tgctaaagcg
     5701 attctttctg cacgattgag taaatcaaga cgattagaaa atctcattgc tcagctcccc
     5761 ggtgagaaga aaaatggctt atttgggaat ctcattgctt tgtcattggg tttgacccct
     5821 aattttaaat caaattttga tttggcagaa gatgctaaat tacagctttc aaaagatact
     5881 tacgatgatg atttagataa tttattggcg caaattggag atcaatatgc tgatttgttt
     5941 ttggcagcta agaatttatc agatgctatt ttactttcag atatcctaag agtaaatact
     6001 gaaataacta aggctcccct atcagcttca atgattaaac gctacgatga acatcatcaa
     6061 gacttgactc ttttaaaagc tttagttcga caacaacttc cagaaaagta taaagaaatc
     6121 ttttttgatc aatcaaaaaa cggatatgca ggttatattg atgggggagc tagccaagaa
     6181 gaattttata aatttatcaa accaatttta gaaaaaatgg atggtactga ggaattattg
     6241 gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa cggctctatt
     6301 ccccatcaaa ttcacttggg tgagctgcat gctattttga gaagacaaga agacttttat
     6361 ccatttttaa aagacaatcg tgagaagatt gaaaaaatct tgacttttcg aattccttat
     6421 tatgttggtc cattggcgcg tggcaatagt cgttttgcat ggatgactcg gaagtctgaa
     6481 gaaacaatta ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc agctcaatca
     6541 tttattgaac gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt actaccaaaa
     6601 catagtttgc tttatgagta ttttacggtt tataacgaat tgacaaaggt caaatatgtt
     6661 actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc cattgttgat
     6721 ttactcttca aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa
     6781 aaaatagaat gttttgatag tgttgaaatt tcaggagttg aagatagatt taatgcttca
     6841 ttaggtacct accatgattt gctaaaaatt attaaagata aagatttttt ggataatgaa
     6901 gaaaatgaag atatcttaga ggatattgtt ttaacattga ccttatttga agatagggag
     6961 atgattgagg aaagacttaa aacatatgct cacctctttg atgataaggt gatgaaacag
     7021 cttaaacgtc gccgttatac tggttgggga cgtttgtctc gaaaattgat taatggtatt
     7081 agggataagc aatctggcaa aacaatatta gattttttga aatcagatgg ttttgccaat
     7141 cgcaatttta tgcagctgat ccatgatgat agtttgacat ttaaagaaga cattcaaaaa
     7201 gcacaagtgt ctggacaagg cgatagttta catgaacata ttgcaaattt agctggtagc
     7261 cctgctatta aaaaaggtat tttacagact gtaaaagttg ttgatgaatt ggtcaaagta
     7321 atggggcggc ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact
     7381 caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa
     7441 ttaggaagtc agattcttaa agagcatcct gttgaaaata ctcaattgca aaatgaaaag
     7501 ctctatctct attatctcca aaatggaaga gacatgtatg tggaccaaga attagatatt
     7561 aatcgtttaa gtgattatga tgtcgatgcc attgttccac aaagtttcct taaagacgat
     7621 tcaatagaca ataaggtctt aacgcgttct gataaaaatc gtggtaaatc ggataacgtt
     7681 ccaagtgaag aagtagtcaa aaagatgaaa aactattgga gacaacttct aaacgccaag
     7741 ttaatcactc aacgtaagtt tgataattta acgaaagctg aacgtggagg tttgagtgaa
     7801 cttgataaag ctggttttat caaacgccaa ttggttgaaa ctcgccaaat cactaagcat
     7861 gtggcacaaa ttttggatag tcgcatgaat actaaatacg atgaaaatga taaacttatt
     7921 cgagaggtta aagtgattac cttaaaatct aaattagttt ctgacttccg aaaagatttc
     7981 caattctata aagtacgtga gattaacaat taccatcatg cccatgatgc gtatctaaat
     8041 gccgtcgttg gaactgcttt gattaagaaa tatccaaaac ttgaatcgga gtttgtctat
     8101 ggtgattata aagtttatga tgttcgtaaa atgattgcta agtctgagca agaaataggc
     8161 aaagcaaccg caaaatattt cttttactct aatatcatga acttcttcaa aacagaaatt
     8221 acacttgcaa atggagagat tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga
     8281 gaaattgtct gggataaagg gcgagatttt gccacagtgc gcaaagtatt gtccatgccc
     8341 caagtcaata ttgtcaagaa aacagaagta cagacaggcg gattctccaa ggagtcaatt
     8401 ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa aagactggga tccaaaaaaa
     8461 tatggtggtt ttgatagtcc aacggtagct tattcagtcc tagtggttgc taaggtggaa
     8521 aaagggaaat cgaagaagtt aaaatccgtt aaagagttac tagggatcac aattatggaa
     8581 agaagttcct ttgaaaaaaa tccgattgac tttttagaag ctaaaggata taaggaagtt
     8641 aaaaaagact taatcattaa actacctaaa tatagtcttt ttgagttaga aaacggtcgt
     8701 aaacggatgc tggctagtgc cggagaatta caaaaaggaa atgagctggc tctgccaagc
     8761 aaatatgtga attttttata tttagctagt cattatgaaa agttgaaggg tagtccagaa
     8821 gataacgaac aaaaacaatt gtttgtggag cagcataagc attatttaga tgagattatt
     8881 gagcaaatca gtgaattttc taagcgtgtt attttagcag atgccaattt agataaagtt
     8941 cttagtgcat ataacaaaca tagagacaaa ccaatacgtg aacaagcaga aaatattatt
     9001 catttattta cgttgacgaa tcttggagct cccgctgctt ttaaatattt tgatacaaca
     9061 attgatcgta aacgatatac gtctacaaaa gaagttttag atgccactct tatccatcaa
     9121 tccatcactg gtctttatga aacacgcatt gatttgagtc agctaggagg tgacaccggt
     9181 gggggtagcg gcggctcggg gggtagtggt ggaagcgggg gttcaaagct tactagtgtt
     9241 accggctatc gtctgtttga aaaagagagc ggatccggtg gaagctggag ccatccgcag
     9301 tttgaaaaat aaactagtgt cttcacactc gaagatttcg ttggggactg ggaacagaca
     9361 gccgcctaca acctggacca agtccttgaa cagggaggtg tgtccagttt gctgcagaat
     9421 ctcgccgtgt ccgtaactcc gatccaaagg attgtccgga gcggtgaaaa tgccctgaag
     9481 atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg ccgaccaaat ggcccagatc
     9541 gaagaggtgt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgccc
     9601 tatggcacac tggtaatcga cggggttacg ccgaacatgc tgaactattt cggacggccg
     9661 tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac
     9721 ggcaacaaaa ttatcgacga gcgcctgatc acccccgacg gctccatgct gttccgagta
     9781 accatcaaca gcggtggaag ctggagccat ccgcagtttg aaaaataact cgagcaccac
     9841 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     9901 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     9961 tttttgctga aaggaggaac tatatccgga t
//
