LOCUS       Exported               11297 bp DNA     circular SYN 30-JUL-2022
DEFINITION  Expresses HiFiCas9 nuclease-T4pol.
ACCESSION   .
VERSION     .
KEYWORDS    pX330A-1x6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   
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FEATURES             Location/Qualifiers
     source          1..11297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..293
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        295..572
                     /label=chicken beta-actin promoter
     intron          573..801
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      813..822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             819..887
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     CDS             894..914
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(894..914)
                     /label=Cas9_BamHI_Primer R2
     CDS             939..5039
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANANFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     misc_feature    2790..3725
                     /label=RE cuts
     misc_feature    3006..3008
                     /label=R691A
     primer_bind     complement(3656..3677)
                     /label=Cas9 variants Primer R
     misc_feature    3658..3715
                     /label=Cas9 variants ssDNA pair
     variation       3678..3680
                     /label=G915F
     variation       3681..3683
                     /label=F916P, Delta-F916
     variation       3687..3689
                     /label=R918A
     variation       3690..3692
                     /label=R919P
     variation       3693..3695
                     /label=Q920P
     primer_bind     3696..3715
                     /label=Cas9 variants Primer 1
     CDS             5040..5087
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5066..5088)
                     /label=Cas9_BamHI_Primer R
     misc_feature    5088..5123
                     /label=SG3-linker
     CDS             5124..7814
                     /codon_start=1
                     /label=T4 DNA polymerase
                     /translation="KEFYISIETVGNNIVERYIDENGKERTREVEYLPTMFRHCKEESK
                     YKDIYGKNCAPQKFPSMKDARDWMKRMEDIGLEALGMNDFKLAYISDTYGSEIVYDRKF
                     VRVANCDIEVTGDKFPDPMKAEYEIDAITHYDSIDDRFYVFDLLNSMYGSVSKWDAKLA
                     AKLDCEGGDEVPQEILDRVIYMPFDNERDMLMEYINLWEQKRPAIFTGWNIEGFDVPYI
                     MNRVKMILGERSMKRFSPIGRVKSKLIQNMYGSKEIYSIDGVSILDYLDLYKKFAFTNL
                     PSFSLESVAQHETKKGKLPYDGPINKLRETNHQRYISYNIIDVESVQAIDKIRGFIDLV
                     LSMSYYAKMPFSGVMSPIKTWDAIIFNSLKGEHKVIPQQGSHVKQSFPGAFVFEPKPIA
                     RRYIMSFDLTSLYPSIIRQVNISPETIRGQFKVHPIHEYIAGTAPKPSDEYSCSPNGWM
                     YDKHQEGIIPKEIAKVFFQRKDWKKKMFAEEMNAEAIKKIIMKGAGSCSTKPEVERYVK
                     FSDDFLNELSNYTESVLNSLIEECEKAATLANTNQLNRKILINSLYGALGNIHFRYYDL
                     RNATAITIFGQVGIQWIARKINEYLNKVCGTNDEDFIAAGDTDSVYVCVDKVIEKVGLD
                     RFKEQNDLVEFMNQFGKKKMEPMIDVAYRELCDYMNNREHLMHMDREAISCPPLGSKGV
                     GGFWKAKKRYALNVYDMEDKRFAEPHLKIMGMETQQSSTPKAVQEALEESIRRILQEGE
                     ESVQEYYKNFEKEYRQLDYKVIAEVKTANDIAKYDDKGWPGFKCPFHIRGVLTYRRAVS
                     GLGVAPILDGNKVMVLPLREGNPFGDKCIAWPSGTELPKEIRSDVLSWIDHSTLFQKSF
                     VKPLAGMCESAGMDYEEKASLDFLFG"
     primer_bind     complement(7814..7847)
                     /label=Cas9_BamHI_Primer R
     primer_bind     complement(7824..7877)
                     /label=Cas9_BamHI_Primer R2
     CDS             7824..7844
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7867..7884)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7873..8080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8089..8229
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      8304..8759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8391..8410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8601..8622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8776..8795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8895..8917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8955..8973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9041..9145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9146..10006
                     /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(9364..9383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10177..10765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10666..10685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10840..11145)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     10995..11017
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11009..11026
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11010..11026
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11036..11055
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11036..11054
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11102..11122)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11102..11120)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11141..11157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11141..11157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11154..11176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11165..11181
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(11189..11219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11234..11255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gagtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      181 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
      241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
      301 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
      361 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
      421 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
      481 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
      541 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc
      601 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      661 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
      721 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
      781 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg
      841 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
      901 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
      961 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     1021 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     1081 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     1141 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     1201 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     1261 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     1321 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     1381 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     1441 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     1501 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     1561 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     1621 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     1681 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     1741 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     1801 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     1861 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     1921 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     1981 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     2041 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     2101 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     2161 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     2221 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     2281 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     2341 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     2401 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     2461 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     2521 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     2581 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     2641 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     2701 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     2761 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     2821 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     2881 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     2941 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     3001 ccaacgccaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     3061 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     3121 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     3181 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     3241 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     3301 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     3361 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     3421 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     3481 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     3541 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     3601 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     3661 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     3721 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     3781 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     3841 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     3901 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     3961 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     4021 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     4081 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     4141 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     4201 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     4261 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     4321 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     4381 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     4441 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     4501 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     4561 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     4621 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     4681 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     4741 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     4801 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     4861 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     4921 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     4981 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     5041 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagggc ggcggatctg
     5101 gtggtgggtc tggcggaggc tcgaaggaat tctatattag tatcgagaca gtgggaaaca
     5161 acattgtaga gcggtatatc gatgaaaacg gcaaagaaag gactcgagag gtcgaatacc
     5221 ttccgaccat gtttcgccac tgtaaagaag aatctaagta caaagatatt tacgggaaga
     5281 attgcgctcc ccaaaaattt ccctccatga aggatgctcg agactggatg aagcgcatgg
     5341 aggacatagg tctcgaagca ttggggatga acgattttaa gttggcttac atctccgaca
     5401 cttacgggtc agaaatagtg tacgatagga agttcgttcg cgtggcaaat tgcgatatag
     5461 aggtcactgg agataagttc ccggacccga tgaaggcgga gtacgaaatt gacgctataa
     5521 cacactatga ctcaatcgac gaccggttct atgtatttga cctgctcaat tccatgtacg
     5581 ggtctgtaag caagtgggac gctaaactcg cggctaaact tgactgtgaa ggaggggatg
     5641 aagtacctca ggaaatcttg gacagggtaa tctacatgcc ctttgacaat gaacgagata
     5701 tgcttatgga gtacattaat ttgtgggagc aaaagcgccc cgcaatattt acaggctgga
     5761 acatagaagg gttcgatgta ccgtatatta tgaatcgggt aaagatgatc ctcggagaga
     5821 gaagcatgaa aagattttca cctattggca gagtgaaatc taagttgata caaaacatgt
     5881 atggctcaaa agagatctat tcaatagatg gagttagcat actcgactac ctggatctgt
     5941 ataaaaagtt tgcttttacc aacttgccta gcttctccct tgaaagtgtc gcccaacacg
     6001 agaccaagaa aggtaagctg ccgtacgatg gcccgattaa taaactgcgc gagaccaacc
     6061 atcaaagata tattagctac aacataattg atgtcgaatc tgtgcaagcc attgataaga
     6121 taaggggctt tatcgacctt gtcctgtcaa tgtcctatta cgccaaaatg ccgttctcag
     6181 gtgtaatgtc acccataaag acgtgggatg cgatcatctt caattctctc aagggagagc
     6241 acaaggtgat cccccaacag gggtcccacg taaagcagtc cttcccggga gcttttgtct
     6301 ttgagccaaa gccgatcgcc cgaaggtata tcatgtcttt cgaccttacg tcactttacc
     6361 cttcaattat tcgacaagtg aatatatcac ccgagactat ccggggccag tttaaggtac
     6421 acccaatcca tgagtacata gccggtacag ccccaaaacc cagtgacgaa tactcttgca
     6481 gccctaatgg gtggatgtac gacaaacacc aggagggcat aatcccaaag gaaatcgcga
     6541 aagtattttt ccaacggaaa gactggaaga aaaagatgtt cgcggaggaa atgaacgccg
     6601 aggctattaa aaaaatcatt atgaagggag cgggtagctg ttctaccaag ccagaggtag
     6661 agcgctacgt caaattcagt gatgacttcc ttaatgagct gagtaactac acagagtctg
     6721 tactgaactc actgattgag gaatgtgaaa aagccgcaac acttgctaat accaatcaac
     6781 tgaatcggaa gatcctgatt aattcactgt atggcgcctt gggcaacatt catttcagat
     6841 actacgacct caggaatgcc acggccatta caattttcgg tcaggtcggg atccagtgga
     6901 tcgcccgaaa aatcaacgag tacctcaata aagtgtgtgg taccaatgac gaggatttta
     6961 tcgcagcagg cgataccgat agcgtgtatg tttgcgtcga caaggtcatt gaaaaggtag
     7021 ggctggatcg gtttaaggag cagaatgatc ttgtcgagtt tatgaaccag tttggtaaaa
     7081 aaaagatgga accgatgata gatgtagcgt accgagaact ttgtgactac atgaataatc
     7141 gcgagcactt gatgcacatg gacagggaag cgatttcatg ccccccactc ggttcaaagg
     7201 gcgtaggggg tttctggaaa gctaagaaac ggtacgccct caacgtctat gacatggaag
     7261 acaagaggtt cgcggaacct catttgaaga taatggggat ggagacgcaa cagtcctcaa
     7321 ctccaaaggc tgtgcaagag gctctggaag aaagcatacg acgcatactc caggaggggg
     7381 aagagagtgt tcaggagtat tataaaaact ttgaaaagga gtaccgccag cttgactaca
     7441 aggtaatcgc ggaggttaag actgcgaatg atatcgccaa atatgatgat aaaggatggc
     7501 ccggtttcaa atgccctttc catatacgag gggtcctcac ctaccgccgc gccgtgtctg
     7561 gtctgggggt cgcaccaatt ctcgacggaa ataaggttat ggtactccca ctccgcgagg
     7621 ggaatccgtt tggtgacaaa tgcatcgcct ggccgtctgg tacggagctc cccaaggaaa
     7681 tacgcagcga cgtcctcagt tggatcgacc actccacact gtttcagaag tcattcgtta
     7741 aacctctggc cgggatgtgt gaatccgcgg gtatggacta cgaggagaaa gcttcattgg
     7801 actttctttt cgggggtggt tctccaaaga agaagcggaa ggtctagcct agagctcgct
     7861 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     7921 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     7981 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     8041 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     8101 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     8161 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     8221 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     8281 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     8341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     8401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     8461 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     8521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     8581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     8641 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     8701 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     8761 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     8821 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     8881 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     8941 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     9001 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     9061 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     9121 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     9181 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     9241 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     9301 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     9361 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     9421 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     9481 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     9541 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     9601 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     9661 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     9721 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     9781 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     9841 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     9901 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     9961 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
    10021 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
    10081 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
    10141 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
    10201 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
    10261 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
    10321 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
    10381 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
    10441 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
    10501 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
    10561 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
    10621 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
    10681 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    10741 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    10801 cttttgctgg ccttttgctc acatgtcaga caaatggctc tagagaccgg cgccgctaca
    10861 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
    10921 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
    10981 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
    11041 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt
    11101 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
    11161 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
    11221 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
    11281 tccaccggtg gtctcta
//
