LOCUS       sequence-262117-       10689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 with an N-terminal HUH-tag called PCV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10689)
  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 10689)
  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: PCV-Cas9
FEATURES             Location/Qualifiers
     source          1..10689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             734..754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             767..784
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      852..899
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1090..1107
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1174..1221
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1258..1713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1345..1364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1555..1576
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1739..1843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1844..1912,1913..2704)
                     /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(2062..2081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2875..3463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3364..3383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3617..3634
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3649..3788
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3714..3733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3874..3896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3890..4081)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4890..5972)
                     /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     5934..5953
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5973..6050)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6256..6275
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6359..6378
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6359..6377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6378..6402
                     /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             6417..6439
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6458..6475
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6488..6778
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             7265..7285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7295..10689,1..709)
                     /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             join(7295..10689,1..709)
                     /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"
ORIGIN
        1 cggttttgat agtccgacgg ttgcatattc cgtcctggtc gtggctaaag tcgaaaaagg
       61 taaaagtaaa aaactgaaat ccgtgaaaga actgctgggc attaccatca tggaacgtag
      121 ctcttttgag aaaaacccga ttgacttcct ggaagccaaa ggttacaaag aagtgaaaaa
      181 agatctgatc atcaaactgc cgaaatatag cctgttcgaa ctggaaaacg gccgtaaacg
      241 catgctggca tctgctggtg aactgcagaa aggcaatgaa ctggcactgc cgagtaaata
      301 tgttaacttt ctgtacctgg ctagccatta tgaaaaactg aaaggttctc cggaagataa
      361 cgaacagaaa caactgttcg tcgaacaaca taaacactac ctggatgaaa tcatcgaaca
      421 gatctcagaa ttctcgaaac gcgtgattct ggcggatgcc aatctggaca aagttctgag
      481 cgcgtataac aaacatcgtg ataaaccgat tcgcgaacag gccgaaaata ttatccacct
      541 gtttaccctg acgaacctgg gcgcaccggc agcttttaaa tacttcgata ccacgatcga
      601 ccgtaaacgc tatacctcaa cgaaagaagt tctggatgct accctgattc atcaatcgat
      661 caccggtctg tatgaaacgc gtattgatct gagtcagctg ggcggtgacg gcggtggctc
      721 cggcacccgt ctgccgaaaa agaaacgcaa agtgggcggt ggctcgcatc atcatcatca
      781 tcactaagga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      841 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      901 tgaaaggagg aactatatcc ggatatcccg caagaggccc ggcagtaccg gcataaccaa
      961 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     1021 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     1081 cttgtcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     1141 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1201 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     1261 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1321 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1381 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1441 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1501 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1561 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1621 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1681 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1741 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1801 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1861 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1921 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1981 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2041 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2101 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2161 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2221 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2281 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2341 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     2401 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2461 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2521 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2581 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2641 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2701 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2761 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2821 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3001 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3541 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3601 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3661 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     3721 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3781 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3841 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3901 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3961 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4021 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4081 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     4141 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     4201 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     4261 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     4321 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     4381 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     4441 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     4501 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     4561 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4621 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4681 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4741 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4801 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4861 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4921 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4981 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5041 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5101 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5161 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5221 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5281 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5341 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5401 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5461 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5521 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5581 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5641 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5701 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     5761 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     5821 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     5881 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     5941 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6001 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6061 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6121 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6181 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6241 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6301 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     6361 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
     6421 tttaacttta agaaggagat ataccatgcg tggttctcac caccaccacc accatatggc
     6481 tagcggatcg gactcagaag tcaatcaaga agctaagcca gaggtcaagc cagaagtcaa
     6541 gcctgagact cacatcaatt taaaggtgtc cgatggatct tcagagatct tcttcaagat
     6601 caaaaagacc actcctttaa gaaggctgat ggaagcgttc gctaaaagac agggtaagga
     6661 aatggactcc ttaagattct tgtacgacgg tattagaatt caagctgatc agacccctga
     6721 agatttggac atggaggata acgatattat tgaggctcac agagaacaga ttggtggatc
     6781 cccgagcaaa aaaaacggtc gtagcggtcc gcagccgcat aaacgttggg tttttaccct
     6841 gaataatccg agcgaagatg agcgcaaaaa aatccgtgat ctgccgatta gcctgttcga
     6901 ttatttcatt gttggtgaag aaggtaacga agaaggtcgt acaccgcatc tgcagggttt
     6961 tgcaaatttt gttaaaaaac agacctttaa caaagtgaaa tggtatctgg gtgcccgttg
     7021 ccatattgaa aaagcaaaag gcaccgatca gcagaacaaa gaatattgta gcaaagaagg
     7081 caacctgctg atggaatgtg gtgcaccgcg tagccagggt cagcgtgagc tcgcggaggc
     7141 ggcagctaag gaggctgccg caaaagaggc cgcagcaaag gaggcagctg cgaaagccgc
     7201 agaagcggcg gcaaaagagg ctgctgcaaa agaggcagca gcaaaagaag ctgcggcgaa
     7261 agcaccgaaa aagaaacgca aagtgggctc gagcatggat aaaaaataca gcattggtct
     7321 ggacattggc acgaatagcg ttggttgggc agtgattacc gatgaataca aagtcccgtc
     7381 gaaaaaattc aaagtgctgg gtaacaccga tcgccatagc attaagaaaa acctgatcgg
     7441 tgcgctgctg tttgattctg gcgaaaccgc ggaagcaacg cgtctgaaac gtaccgcacg
     7501 tcgccgttac acgcgccgta aaaatcgtat ttgctatctg caggaaatct ttagcaacga
     7561 aatggcgaaa gtcgatgact catttttcca ccgcctggaa gaatcgtttc tggtggaaga
     7621 agataaaaaa catgaacgtc acccgatttt cggcaatatc gttgatgaag tcgcgtacca
     7681 tgaaaaatat ccgacgattt accacctgcg taaaaaactg gtggattcta ccgacaaagc
     7741 cgatctgcgc ctgatttatc tggcactggc tcatatgatc aaatttcgtg gtcacttcct
     7801 gattgaaggc gacctgaacc cggataatag tgacgtcgat aaactgttta ttcagctggt
     7861 gcaaacctat aatcagctgt tcgaagaaaa cccgatcaat gcaagtggtg ttgatgcgaa
     7921 agccattctg tccgctcgcc tgagtaaatc ccgccgtctg gaaaacctga ttgcacagct
     7981 gccgggtgaa aagaaaaacg gtctgtttgg caatctgatc gctctgtcac tgggcctgac
     8041 gccgaacttt aaatcgaatt tcgacctggc agaagatgct aaactgcagc tgagcaaaga
     8101 tacctacgat gacgatctgg acaacctgct ggcgcaaatt ggcgaccagt atgccgacct
     8161 gtttctggcg gccaaaaatc tgtcagatgc cattctgctg tcggacatcc tgcgcgtgaa
     8221 caccgaaatc acgaaagcgc cgctgtcagc ctcgatgatt aaacgctacg atgaacatca
     8281 ccaggacctg accctgctga aagcactggt tcgtcagcaa ctgccggaaa aatacaaaga
     8341 aattttcttt gaccaaagta aaaatggtta tgcaggctac atcgatggcg gtgcttccca
     8401 ggaagaattc tacaaattca tcaaaccgat cctggaaaaa atggatggta cggaagaact
     8461 gctggtgaaa ctgaatcgtg aagatctgct gcgtaaacaa cgcacctttg acaacggtag
     8521 cattccgcat cagatccacc tgggcgaact gcatgcgatt ctgcgccgtc aggaagattt
     8581 ttatccgttc ctgaaagaca accgtgaaaa aatcgaaaaa atcctgacgt ttcgcatccc
     8641 gtattacgtt ggtccgctgg cacgtggtaa tagccgcttc gcatggatga cccgcaaatc
     8701 tgaagaaacc attacgccgt ggaactttga agaagtggtt gataaaggcg caagcgctca
     8761 gtcttttatc gaacgtatga ccaatttcga taaaaacctg ccgaatgaaa aagtgctgcc
     8821 gaaacattct ctgctgtatg aatactttac cgtttacaac gaactgacga aagtgaaata
     8881 tgttaccgag ggtatgcgca aaccggcgtt tctgagtggc gaacagaaaa aagccattgt
     8941 ggatctgctg ttcaaaacca atcgtaaagt tacggtcaaa cagctgaaag aagattactt
     9001 caagaaaatt gaatgtttcg acagcgtgga aatttctggt gttgaagatc gtttcaacgc
     9061 ctctctgggc acctatcatg acctgctgaa aatcatcaaa gacaaagatt ttctggataa
     9121 cgaagaaaac gaagacattc tggaagatat cgtgctgacc ctgacgctgt tcgaagatcg
     9181 tgaaatgatt gaagaacgcc tgaaaacgta cgcacacctg tttgacgata aagttatgaa
     9241 acagctgaaa cgccgtcgct ataccggttg gggccgtctg agccgcaaac tgattaatgg
     9301 tatccgcgat aaacaatcag gcaaaacgat tctggatttc ctgaaatcgg acggctttgc
     9361 caaccgtaat ttcatgcagc tgatccatga cgattccctg acctttaaag aagacattca
     9421 gaaagcacaa gtgtcaggtc aaggcgattc gctgcatgaa cacattgcga acctggccgg
     9481 ttcaccggct atcaaaaaag gcatcctgca gaccgtgaaa gtcgtggatg aactggtgaa
     9541 agttatgggt cgtcacaaac cggaaaacat tgttatcgaa atggcgcgcg aaaatcagac
     9601 cacgcaaaaa ggccagaaaa actcgcgtga acgcatgaaa cgcattgaag aaggtatcaa
     9661 agaactgggc agccagattc tgaaagaaca tccggtcgaa aacacccagc tgcaaaatga
     9721 aaaactgtac ctgtattacc tgcaaaatgg tcgtgacatg tatgtggatc aggaactgga
     9781 catcaaccgc ctgtctgact atgatgtcga ccacattgtg ccgcagagct ttctgaaaga
     9841 cgattctatc gataacaaag ttctgacccg tagtgataaa aaccgcggca aaagcgacaa
     9901 tgtcccgtct gaagaagttg tgaagaaaat gaaaaactac tggcgtcaac tgctgaatgc
     9961 gaaactgatt acgcagcgta aattcgataa cctgaccaaa gcggaacgcg gcggtctgtc
    10021 cgaactggat aaagccggtt ttatcaaacg tcaactggtt gaaacccgcc agattacgaa
    10081 acatgtcgcc cagatcctgg attcacgcat gaacacgaaa tacgacgaaa acgataaact
    10141 gatccgtgaa gtcaaagtga tcaccctgaa aagtaaactg gtttccgatt tccgtaaaga
    10201 ctttcagttc tacaaagtcc gcgaaattaa caattaccat cacgcacacg atgcttatct
    10261 gaatgcagtg gttggtaccg ctctgatcaa aaaatatccg aaactggaaa gcgaatttgt
    10321 gtatggcgat tacaaagtct atgacgtgcg caaaatgatt gcgaaatccg aacaggaaat
    10381 cggcaaagcg accgccaaat actttttcta ttcaaacatc atgaactttt tcaaaaccga
    10441 aattacgctg gcaaatggtg aaattcgtaa acgcccgctg atcgaaacca acggtgaaac
    10501 gggcgaaatt gtgtgggata aaggccgtga cttcgcgacc gttcgcaaag tcctgtcgat
    10561 gccgcaagtg aatatcgtga agaaaaccga agtgcagacg ggcggtttta gtaaagaatc
    10621 catcctgccg aaacgtaaca gcgataaact gattgcgcgc aaaaaagatt gggacccgaa
    10681 aaaatacgg
//
