LOCUS       sequence-314790-       11255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of dCas9-BirA*.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11255)
  AUTHORS   Wang AS, Chen LC, Wu RA, Hao Y, McSwiggen DT, Heckert AB, Richardson
            CD, Gowen BG, Kazane KR, Vu JT, Wyman SK, Shin JJ, Darzacq X, Walter
            JC, Corn JE
  TITLE     The Histone Chaperone FACT Induces Cas9 Multi-turnover Behavior and 
            Modifies Genome Manipulation in Human Cells.
  JOURNAL   Mol Cell. 2020 Jul 16;79(2):221-233.e5. doi: 
            10.1016/j.molcel.2020.06.014. Epub 2020 Jun 29.
  PUBMED    32603710
REFERENCE   2  (bases 1 to 11255)
  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: 
            "10.1016/j.molcel.2020.06.014"; journalName: "Mol Cell"; date: 
            "2020-07-16- 16"; volume: "79"; issue: "2"; pages: "221-233.e5"
COMMENT     Sequence Label: T7pr-His6-MBP-TEV-dCas9-BirA*
FEATURES             Location/Qualifiers
     source          1..11255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(106..174,175..966)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(324..343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1137..1725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1626..1645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1879..1896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1911..2050
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1976..1995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2136..2158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2152..2343)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3801..3820
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3904..3923
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3904..3922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4011..4033
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4053..4070
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4080..5180
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     5154..5177
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             5235..5255
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      5259..5268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5268..5333
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5340..5360
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5385..5408,5409..5411,5412..7898,7899..7901,
                     7902..9485)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5385..9485
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9486..9533
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(9570..9917,9918..9920,9921..10529)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             10533..10559
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(10688..10706)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      10702..10749
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(11115..11120,11121..11137,11138..11143))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(11170..11188)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataacat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     1921 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     1981 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     2041 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2101 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2161 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     2221 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     2281 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     2341 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     2401 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     2461 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     2521 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     2581 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     2641 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     2701 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     2761 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
     2821 aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     2881 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     2941 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg
     3001 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc
     3061 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt
     3121 ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg
     3181 tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa
     3241 gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg
     3301 tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     3361 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     3421 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     3481 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     3541 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3601 ggtcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     3661 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     3721 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     3781 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     3841 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     3901 aattaatacg actcactata gggagaccac aacggtttcc ctctagtgcc ggctccggag
     3961 agctctttaa ttaagcggcc gccctgcagg actcgagttc tagaaataat tttgtttaac
     4021 tttaagaagg agatatacat atgaaatctt ctcaccatca ccatcaccat ggttcttcta
     4081 tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc
     4141 tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc
     4201 cggataaact ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta
     4261 tcttctgggc acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca
     4321 ccccggacaa agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca
     4381 acggcaagct gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag
     4441 atctgctgcc gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga
     4501 aagcgaaagg taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc
     4561 tgattgctgc tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag
     4621 acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta
     4681 aaaacaaaca catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag
     4741 gcgaaacagc gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag
     4801 tgaattatgg tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg
     4861 gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc
     4921 tcgaaaacta tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg
     4981 gtgccgtagc gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca
     5041 ctatggaaaa cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct
     5101 ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag
     5161 ccctgaaaga cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca
     5221 acctcgggat cgaggaaaac ctgtacttcc aatccaatgc caccatggac tataaggacc
     5281 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc
     5341 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca
     5401 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     5461 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     5521 tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg ctgaagagaa
     5581 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     5641 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     5701 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     5761 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     5821 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     5881 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     5941 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     6001 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     6061 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg
     6121 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     6181 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     6241 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     6301 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     6361 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     6421 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc gatggcggag
     6481 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     6541 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     6601 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     6661 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     6721 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     6781 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgcca
     6841 gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     6901 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag
     6961 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaaag
     7021 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     7081 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     7141 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     7201 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     7261 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     7321 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     7381 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     7441 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     7501 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     7561 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     7621 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     7681 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     7741 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     7801 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     7861 aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc
     7921 tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac cggggcaaga
     7981 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cgccagctgc
     8041 tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc gagagaggcg
     8101 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     8161 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaacg
     8221 acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     8281 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     8341 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     8401 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     8461 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     8521 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     8581 gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg cggaaagtgc
     8641 tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     8701 aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag aaggactggg
     8761 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     8821 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     8881 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     8941 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     9001 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     9061 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     9121 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa cactacctgg
     9181 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     9241 tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga gagcaggccg
     9301 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     9361 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     9421 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     9481 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagggcggtg
     9541 gtagcggtgg cggaagcggt ggaggcagta aggacaacac cgtgcccctg aagctgatcg
     9601 ccctgctggc caacggcgag ttccactctg gcgagcagct gggagagacc ctgggaatga
     9661 gcagagccgc catcaacaag cacatccaga cactgagaga ctggggagtg gacgtgttca
     9721 ccgtgcctgg caagggctac agcctgcctg agcctatcca gctgctgaac gccaagcaga
     9781 tcctgggaca gctggatggc ggaagcgtgg ccgtgctgcc tgtgatcgac tccaccaatc
     9841 agtacctgct ggacagaatc ggagagctga agtccggcga cgcctgcatc gccgagtacc
     9901 agcaggctgg cagaggaggc agaggacgga agtggttcag cccattcgga gccaacctgt
     9961 acctgtccat gttctggaga ctggagcagg gacctgctgc tgccatcgga ctgagtctgg
    10021 tgatcggaat cgtgatggcc gaggtgctga gaaagctggg agccgacaag gtgagagtga
    10081 agtggcctaa tgacctgtac ctccaggacc gcaagctggc tggcatcctg gtggagctga
    10141 caggcaagac aggcgatgcc gctcagatcg tgatcggagc cggaatcaac atggccatga
    10201 gaagagtgga ggagagcgtg gtgaaccagg gctggatcac cctgcaggag gctggcatca
    10261 acctggaccg gaacaccctg gccgccatgc tgatcagaga gctgagagcc gctctggagc
    10321 tgttcgagca ggagggactg gctccttacc tgagcagatg ggagaagctg gacaacttca
    10381 tcaacagacc tgtgaagctg atcatcggcg acaaggaaat cttcggcatc tccagaggaa
    10441 tcgacaagca gggagctctg ctgctggagc aggacggaat catcaagccc tggatgggcg
    10501 gagaaatctc cctgagaagc gcagagaagg cttaccctta cgatgtaccg gattacgcat
    10561 agtaacattg gaagtggata acggatccgc gatcgcggcg cgccacctgg tggccggccg
    10621 gtaccacgcg tgcgcgctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
    10681 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
    10741 ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat
    10801 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa
    10861 gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc
    10921 tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc
    10981 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
    11041 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
    11101 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
    11161 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
    11221 cttagacgtc aggtggcact tttcggggaa atgtg
//
