LOCUS       sequence-171567-        9418 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpCas9 fused to ER50 domain on both N- and C-termini in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9418)
  AUTHORS   Maji B, Moore CL, Zetsche B, Volz SE, Zhang F, Shoulders MD, 
            Choudhary A
  TITLE     Multidimensional chemical control of CRISPR-Cas9.
  JOURNAL   Nat Chem Biol. 2017 Jan;13(1):9-11. doi: 10.1038/nchembio.2224. Epub
            2016 Oct 31.
  PUBMED    27820801
REFERENCE   2  (bases 1 to 9418)
  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: 
            "10.1038/nchembio.2224"; journalName: "Nat Chem Biol"; date: 
            "2017-01"; volume: "13"; issue: "1"; pages: "9-11"
COMMENT     Sequence Label: ER50-SpCas9-ER50
FEATURES             Location/Qualifiers
     source          1..9418
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      226..235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             235..300
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             307..327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1111..5211
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1111..5211
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5212..5259
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6022..6039)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6028..6235
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6244..6384
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6244..6373
                     /label=AAV2 ITR
     rep_origin      6459..6914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6546..6565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6756..6777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6931..6950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7050..7072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7110..7128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7196..7300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7301..7369,7370..8161)
                     /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     complement(7519..7538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8332..8920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8821..8840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8982..9230
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8982..9002
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9153..9172
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9249..9324
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
       61 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      121 gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta
      181 cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat
      241 aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga tgacgataag
      301 atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc cggatccagc
      361 cttgccctgt cacttacagc cgaccagatg gtttccgcgc ttctcgacgc tgaacctcca
      421 attctctatt ccgaatacga cccaaccagg ccgttctccg aggcatctat gatgggtctg
      481 ctgacaaatc tggcagacag ggaactggtg cacatgatca attgggcgaa gcgcgtaccc
      541 ggattcgtcg atcttgcact ccatgatcag gtgcacttgc tggagtgcgc ttggatggag
      601 atcctcatga tcgggctggt gtggcggagt atggaacacc ccggcaagtt gctgtttgcg
      661 cctaacctcc tgttggacag gaaccagggg aaatgtgtgg agggcggtgt ggaaatcttt
      721 gacatgctcc tcgctacctc aagccggttt aggatgatga atctgcaggg cgaagagttc
      781 gtgtgtctca aatctatcat actgttgaac agcggagtct acaccttcct ctccagtact
      841 ctgaaatctc tggaggagaa agatcatatc catcgcgtgc tggacaagat aaccgacacg
      901 ttgattcact tgatggccaa agctgggctc actctgcaac aacaacatca gcgactggca
      961 cagctgttgc tgattttgag ccacattcgg cacatgtcca gcaagagaat ggagcacctc
     1021 tatagtatga agtgcaagaa cgtcgtaccc ctgtcagatc tgcttcttga aatgcttgat
     1081 gcccaccggc tgggaagcgg aagcggatcc gacaagaagt acagcatcgg cctggacatc
     1141 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     1201 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     1261 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     1321 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     1381 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     1441 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     1501 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     1561 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     1621 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     1681 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     1741 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     1801 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     1861 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     1921 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     1981 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     2041 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     2101 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     2161 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     2221 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     2281 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     2341 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     2401 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     2461 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     2521 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     2581 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     2641 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     2701 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     2761 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     2821 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     2881 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     2941 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     3001 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     3061 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     3121 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     3181 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     3241 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     3301 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     3361 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     3421 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     3481 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     3541 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     3601 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
     3661 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     3721 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     3781 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     3841 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     3901 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
     3961 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     4021 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     4081 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     4141 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     4201 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     4261 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
     4321 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
     4381 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
     4441 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     4501 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     4561 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     4621 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     4681 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     4741 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
     4801 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     4861 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
     4921 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
     4981 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
     5041 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
     5101 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     5161 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga caaaaggccg
     5221 gcggccacga aaaaggccgg ccaggcaaaa aagaaaaaga gccttgccct gtcacttaca
     5281 gccgaccaga tggtttccgc gcttctcgac gctgaacctc caattctcta ttccgaatac
     5341 gacccaacca ggccgttctc cgaggcatct atgatgggtc tgctgacaaa tctggcagac
     5401 agggaactgg tgcacatgat caattgggcg aagcgcgtac ccggattcgt cgatcttgca
     5461 ctccatgatc aggtgcactt gctggagtgc gcttggatgg agatcctcat gatcgggctg
     5521 gtgtggcgga gtatggaaca ccccggcaag ttgctgtttg cgcctaacct cctgttggac
     5581 aggaaccagg ggaaatgtgt ggagggcggt gtggaaatct ttgacatgct cctcgctacc
     5641 tcaagccggt ttaggatgat gaatctgcag ggcgaagagt tcgtgtgtct caaatctatc
     5701 atactgttga acagcggagt ctacaccttc ctctccagta ctctgaaatc tctggaggag
     5761 aaagatcata tccatcgcgt gctggacaag ataaccgaca cgttgattca cttgatggcc
     5821 aaagctgggc tcactctgca acaacaacat cagcgactgg cacagctgtt gctgattttg
     5881 agccacattc ggcacatgtc cagcaagaga atggagcacc tctatagtat gaagtgcaag
     5941 aacgtcgtac ccctgtcaga tctgcttctt gaaatgcttg atgcccaccg gctgtaagaa
     6001 ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6061 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6121 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     6181 tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc
     6241 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     6301 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     6361 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     6421 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     6481 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     6541 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6601 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6661 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6721 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6781 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     6841 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6901 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6961 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     7021 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     7081 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     7141 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     7201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7261 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7321 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7381 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7441 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7501 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7561 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7621 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7681 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7741 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7801 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7861 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7921 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7981 gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact
     8041 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8101 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8161 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8221 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8281 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     8341 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     8401 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     8461 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     8521 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     8581 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     8641 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     8701 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     8761 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     8821 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     8881 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     8941 tttacggttc ctggcctttt gctggccttt tgctcacatg tgagggccta tttcccatga
     9001 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga
     9061 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     9121 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     9181 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gggtcttcga
     9241 gaagacctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     9301 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
     9361 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggta
//
