LOCUS       sequence-484257-        8277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian vector for transient expression of dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8277)
  AUTHORS   Stohr AM, Hansen H, Richards B, Park H, Goncalves AG, Agrawal A, 
            Blenner M, Chen W
  TITLE     Metabolite-responsive scaffold RNAs for dynamic CRISPR 
            transcriptional regulation.
  JOURNAL   Nucleic Acids Res. 2025 Nov 13;53(21):gkaf1290. doi: 
            10.1093/nar/gkaf1290.
  PUBMED    41277686
REFERENCE   2  (bases 1 to 8277)
  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: "Nucleic 
            Acids Res."; date: "2025-11-13"; volume: "53(21):gkaf1290. doi"; 
            pages: " 10.1093/nar/gkaf1290"
COMMENT     Sequence Label: pHS-dCas9
FEATURES             Location/Qualifiers
     source          1..8277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   75..109
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        431..930
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(466..489)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     863..882
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      954..963
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(960..986,987..989,990..3476,3477..3479,3480..5063)
                     /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             960..5063
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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             5067..5093
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5073..5099
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             5112..5132
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5139..5159
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   5377..5439
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5490..5510)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5490..5508)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5529..5545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5529..5545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5542..5564)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5553..5569
                     /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(5577..5583,5584..5601,5602..5607))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5622..5643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5760..5777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5931..6519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6011..6030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6690..7481,7482..7550))
                     /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     7313..7332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7551..7655)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7681..8136)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7818..7839)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8030..8049
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8263..8277,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8277,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggggcgcg
       61 ccattctaga ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa
      121 tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc
      181 gccgcttgga gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac
      241 tataacgacc gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt
      301 aactcatacg ataattatat tgttatttca tgttctactt acgtgataac ttattatata
      361 tatattttct tgttatagat atcaactaga atgctagcat gggcccagat ctcctgcagg
      421 taattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
      481 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
      541 accggtaggc accaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
      601 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
      661 gaagtagcac gtctcactag gctcgtgcag atggacagca ccgctgagca atggaagcgg
      721 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
      781 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
      841 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
      901 ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttgtc gacgccacca
      961 tggacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga
     1021 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     1081 acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa acagccgagg
     1141 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     1201 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     1261 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     1321 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     1381 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     1441 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     1501 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     1561 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     1621 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggcaacc
     1681 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     1741 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1801 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1861 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     1921 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     1981 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     2041 gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     2101 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     2161 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     2221 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     2281 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     2341 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     2401 tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     2461 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     2521 acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     2581 gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     2641 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     2701 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     2761 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     2821 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     2881 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     2941 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     3001 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     3061 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     3121 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     3181 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     3241 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     3301 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     3361 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     3421 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggacgcta
     3481 tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg actcggagcg
     3541 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     3601 actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc gacaatctga
     3661 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     3721 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     3781 ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc ctgaagtcca
     3841 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     3901 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     3961 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     4021 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     4081 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     4141 ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc cgggactttg
     4201 ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     4261 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac aagctgatcg
     4321 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     4381 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     4441 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     4501 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     4561 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     4621 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     4681 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     4741 agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     4801 tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac agagacaagc
     4861 ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     4921 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     4981 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     5041 acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat gccagcctgg
     5101 gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag cggaaagtgg
     5161 acggcattgg tagtgggagc aacggatgac tcgagctagt taaaagtttt gttactttat
     5221 agaagaaatt ttgagttttt gttttttttt taataaataa ataaacataa ataaattgtt
     5281 tgttgaattt attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat
     5341 taataaataa acctcgatat acagaccgat aaaacacatg cgtcaatttt acgcatgatt
     5401 atctttaacg tacgtcacaa tatgattatc tttctagggt taatctagta tacgcgtatg
     5461 cggccgctta attaatccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     5521 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5581 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5641 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5701 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5761 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5821 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5881 aaaggtcagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5941 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6001 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6061 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6121 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6181 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6241 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6301 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6361 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6421 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6481 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6541 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6601 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6661 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6721 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6781 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6841 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6901 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6961 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7021 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7081 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7141 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7201 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7261 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7321 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7381 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7441 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7501 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7561 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7621 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7681 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     7741 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     7801 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     7861 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     7921 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     7981 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     8041 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     8101 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     8161 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     8221 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
