LOCUS       sequence-239426-       10224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 with a C-terminal HUH-tag called PCV2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10224)
  AUTHORS   Aird EJ, Lovendahl KN, St Martin A, Harris RS, Gordon WR
  TITLE     Increasing Cas9-mediated homology-directed repair efficiency through
            covalent tethering of DNA repair template.
  JOURNAL   Commun Biol. 2018 May 31;1:54. doi: 10.1038/s42003-018-0054-2. 
            eCollection 2018.
  PUBMED    30271937
REFERENCE   2  (bases 1 to 10224)
  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: "Commun 
            Biol."; date: "2018-05-31"; volume: "1:54. doi"; pages: " 
            10.1038/s42003-018-0054-2. eCollection 2018"
COMMENT     Sequence Label: Cas9-PCV
FEATURES             Location/Qualifiers
     source          1..10224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..198)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1007..2089)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2051..2070
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2090..2167)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2373..2392
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2476..2495
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2476..2494
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2495..2519
                     /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             2534..2556
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2587..2877
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             2881..6984
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2881..6984
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7009..7029
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7396..7413
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7431..7448
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7501..7519)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7515..7562
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      7599..8054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7686..7705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7896..7917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8080..8184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8185..8253,8254..9045)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8403..8422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9216..9804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9705..9724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9958..9975
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9990..10129
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10055..10074)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
       61 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
      121 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
      181 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
      241 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
      301 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
      361 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
      421 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
      481 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
      541 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
      601 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
      661 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
      721 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
      781 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
      841 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
      901 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
      961 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     1021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1081 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgatacg ggcaacagct
     1141 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     1201 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     1261 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     1321 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     1381 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     1441 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     1501 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     1561 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     1621 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     1681 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     1741 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     1801 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     1861 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     1921 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     1981 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     2041 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     2101 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     2161 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     2221 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     2281 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     2341 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     2401 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     2461 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     2521 ctctagaaat aattttgttt aactttaaga aggagatata ccatgcgtgg ttctatggct
     2581 agcggatcgg actcagaagt caatcaagaa gctaagccag aggtcaagcc agaagtcaag
     2641 cctgagactc acatcaattt aaaggtgtcc gatggatctt cagagatctt cttcaagatc
     2701 aaaaagacca ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca gggtaaggaa
     2761 atggactcct taagattctt gtacgacggt attagaattc aagctgatca gacccctgaa
     2821 gatttggaca tggaggataa cgatattatt gaggctcaca gagaacagat tggtggatcc
     2881 atggataaaa aatacagcat tggtctggac attggcacga atagcgttgg ttgggcagtg
     2941 attaccgatg aatacaaagt cccgtcgaaa aaattcaaag tgctgggtaa caccgatcgc
     3001 catagcatta agaaaaacct gatcggtgcg ctgctgtttg attctggcga aaccgcggaa
     3061 gcaacgcgtc tgaaacgtac cgcacgtcgc cgttacacgc gccgtaaaaa tcgtatttgc
     3121 tatctgcagg aaatctttag caacgaaatg gcgaaagtcg atgactcatt tttccaccgc
     3181 ctggaagaat cgtttctggt ggaagaagat aaaaaacatg aacgtcaccc gattttcggc
     3241 aatatcgttg atgaagtcgc gtaccatgaa aaatatccga cgatttacca cctgcgtaaa
     3301 aaactggtgg attctaccga caaagccgat ctgcgcctga tttatctggc actggctcat
     3361 atgatcaaat ttcgtggtca cttcctgatt gaaggcgacc tgaacccgga taatagtgac
     3421 gtcgataaac tgtttattca gctggtgcaa acctataatc agctgttcga agaaaacccg
     3481 atcaatgcaa gtggtgttga tgcgaaagcc attctgtccg ctcgcctgag taaatcccgc
     3541 cgtctggaaa acctgattgc acagctgccg ggtgaaaaga aaaacggtct gtttggcaat
     3601 ctgatcgctc tgtcactggg cctgacgccg aactttaaat cgaatttcga cctggcagaa
     3661 gatgctaaac tgcagctgag caaagatacc tacgatgacg atctggacaa cctgctggcg
     3721 caaattggcg accagtatgc cgacctgttt ctggcggcca aaaatctgtc agatgccatt
     3781 ctgctgtcgg acatcctgcg cgtgaacacc gaaatcacga aagcgccgct gtcagcctcg
     3841 atgattaaac gctacgatga acatcaccag gacctgaccc tgctgaaagc actggttcgt
     3901 cagcaactgc cggaaaaata caaagaaatt ttctttgacc aaagtaaaaa tggttatgca
     3961 ggctacatcg atggcggtgc ttcccaggaa gaattctaca aattcatcaa accgatcctg
     4021 gaaaaaatgg atggtacgga agaactgctg gtgaaactga atcgtgaaga tctgctgcgt
     4081 aaacaacgca cctttgacaa cggtagcatt ccgcatcaga tccacctggg cgaactgcat
     4141 gcgattctgc gccgtcagga agatttttat ccgttcctga aagacaaccg tgaaaaaatc
     4201 gaaaaaatcc tgacgtttcg catcccgtat tacgttggtc cgctggcacg tggtaatagc
     4261 cgcttcgcat ggatgacccg caaatctgaa gaaaccatta cgccgtggaa ctttgaagaa
     4321 gtggttgata aaggcgcaag cgctcagtct tttatcgaac gtatgaccaa tttcgataaa
     4381 aacctgccga atgaaaaagt gctgccgaaa cattctctgc tgtatgaata ctttaccgtt
     4441 tacaacgaac tgacgaaagt gaaatatgtt accgagggta tgcgcaaacc ggcgtttctg
     4501 agtggcgaac agaaaaaagc cattgtggat ctgctgttca aaaccaatcg taaagttacg
     4561 gtcaaacagc tgaaagaaga ttacttcaag aaaattgaat gtttcgacag cgtggaaatt
     4621 tctggtgttg aagatcgttt caacgcctct ctgggcacct atcatgacct gctgaaaatc
     4681 atcaaagaca aagattttct ggataacgaa gaaaacgaag acattctgga agatatcgtg
     4741 ctgaccctga cgctgttcga agatcgtgaa atgattgaag aacgcctgaa aacgtacgca
     4801 cacctgtttg acgataaagt tatgaaacag ctgaaacgcc gtcgctatac cggttggggc
     4861 cgtctgagcc gcaaactgat taatggtatc cgcgataaac aatcaggcaa aacgattctg
     4921 gatttcctga aatcggacgg ctttgccaac cgtaatttca tgcagctgat ccatgacgat
     4981 tccctgacct ttaaagaaga cattcagaaa gcacaagtgt caggtcaagg cgattcgctg
     5041 catgaacaca ttgcgaacct ggccggttca ccggctatca aaaaaggcat cctgcagacc
     5101 gtgaaagtcg tggatgaact ggtgaaagtt atgggtcgtc acaaaccgga aaacattgtt
     5161 atcgaaatgg cgcgcgaaaa tcagaccacg caaaaaggcc agaaaaactc gcgtgaacgc
     5221 atgaaacgca ttgaagaagg tatcaaagaa ctgggcagcc agattctgaa agaacatccg
     5281 gtcgaaaaca cccagctgca aaatgaaaaa ctgtacctgt attacctgca aaatggtcgt
     5341 gacatgtatg tggatcagga actggacatc aaccgcctgt ctgactatga tgtcgaccac
     5401 attgtgccgc agagctttct gaaagacgat tctatcgata acaaagttct gacccgtagt
     5461 gataaaaacc gcggcaaaag cgacaatgtc ccgtctgaag aagttgtgaa gaaaatgaaa
     5521 aactactggc gtcaactgct gaatgcgaaa ctgattacgc agcgtaaatt cgataacctg
     5581 accaaagcgg aacgcggcgg tctgtccgaa ctggataaag ccggttttat caaacgtcaa
     5641 ctggttgaaa cccgccagat tacgaaacat gtcgcccaga tcctggattc acgcatgaac
     5701 acgaaatacg acgaaaacga taaactgatc cgtgaagtca aagtgatcac cctgaaaagt
     5761 aaactggttt ccgatttccg taaagacttt cagttctaca aagtccgcga aattaacaat
     5821 taccatcacg cacacgatgc ttatctgaat gcagtggttg gtaccgctct gatcaaaaaa
     5881 tatccgaaac tggaaagcga atttgtgtat ggcgattaca aagtctatga cgtgcgcaaa
     5941 atgattgcga aatccgaaca ggaaatcggc aaagcgaccg ccaaatactt tttctattca
     6001 aacatcatga actttttcaa aaccgaaatt acgctggcaa atggtgaaat tcgtaaacgc
     6061 ccgctgatcg aaaccaacgg tgaaacgggc gaaattgtgt gggataaagg ccgtgacttc
     6121 gcgaccgttc gcaaagtcct gtcgatgccg caagtgaata tcgtgaagaa aaccgaagtg
     6181 cagacgggcg gttttagtaa agaatccatc ctgccgaaac gtaacagcga taaactgatt
     6241 gcgcgcaaaa aagattggga cccgaaaaaa tacggcggtt ttgatagtcc gacggttgca
     6301 tattccgtcc tggtcgtggc taaagtcgaa aaaggtaaaa gtaaaaaact gaaatccgtg
     6361 aaagaactgc tgggcattac catcatggaa cgtagctctt ttgagaaaaa cccgattgac
     6421 ttcctggaag ccaaaggtta caaagaagtg aaaaaagatc tgatcatcaa actgccgaaa
     6481 tatagcctgt tcgaactgga aaacggccgt aaacgcatgc tggcatctgc tggtgaactg
     6541 cagaaaggca atgaactggc actgccgagt aaatatgtta actttctgta cctggctagc
     6601 cattatgaaa aactgaaagg ttctccggaa gataacgaac agaaacaact gttcgtcgaa
     6661 caacataaac actacctgga tgaaatcatc gaacagatct cagaattctc gaaacgcgtg
     6721 attctggcgg atgccaatct ggacaaagtt ctgagcgcgt ataacaaaca tcgtgataaa
     6781 ccgattcgcg aacaggccga aaatattatc cacctgttta ccctgacgaa cctgggcgca
     6841 ccggcagctt ttaaatactt cgataccacg atcgaccgta aacgctatac ctcaacgaaa
     6901 gaagttctgg atgctaccct gattcatcaa tcgatcaccg gtctgtatga aacgcgtatt
     6961 gatctgagtc agctgggcgg tgacggcggt ggctccggca cccgtctgcc gaaaaagaaa
     7021 cgcaaagtgg gcggtggctc gggatccccg agcaaaaaaa acggtcgtag cggtccgcag
     7081 ccgcataaac gttgggtttt taccctgaat aatccgagcg aagatgagcg caaaaaaatc
     7141 cgtgatctgc cgattagcct gttcgattat ttcattgttg gtgaagaagg taacgaagaa
     7201 ggtcgtacac cgcatctgca gggttttgca aattttgtta aaaaacagac ctttaacaaa
     7261 gtgaaatggt atctgggtgc ccgttgccat attgaaaaag caaaaggcac cgatcagcag
     7321 aacaaagaat attgtagcaa agaaggcaac ctgctgatgg aatgtggtgc accgcgtagc
     7381 cagggtcagc gtggtcacca ccaccaccac cattaaataa gcttgtcgag caccaccacc
     7441 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     7501 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     7561 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     7621 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     7681 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7741 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7801 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7861 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7921 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7981 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     8041 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     8101 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     8161 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     8221 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     8281 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     8341 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     8401 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     8461 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     8521 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     8581 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     8641 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     8701 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     8761 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     8821 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     8881 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     8941 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     9001 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     9061 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     9121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     9181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     9241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     9301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     9361 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     9421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     9481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     9541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     9601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     9661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     9721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     9781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     9841 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     9901 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     9961 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
    10021 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
    10081 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
    10141 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
    10201 agctgtgacc gtctgcggga gctg
//
