p35S-PBE

LOCUS       p35S-PBE               17062 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Conversion of C to T in potato.
ACCESSION   .
VERSION     .
KEYWORDS    p35S-PBE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17062)
  AUTHORS   Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C
  TITLE     Efficient C-to-T base editing in plants using a fusion of nCas9 and 
            human APOBEC3A.
  JOURNAL   Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261.
  PUBMED    30272679
REFERENCE   2  (bases 1 to 17062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..17062
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(919..1548)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    2849..2873
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     3061..3083
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3075..3092
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3076..3092
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3805..4596
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        4748..4823
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5437..5781
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5930..5950
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5951..6634
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             6686..10786
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10787..10834
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10856..11104
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             11117..11137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(11793..11809)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11793..11809)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11806..11828)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11817..11833
                     /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(11841..11871)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11886..11907
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12098..12776
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             12821..13372
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    13379..13553
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    13631..13655
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             14080..14874
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      14961..15549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15450..15469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     15703..15720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    15735..15875
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(15801..15820)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     15961..15983
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      16219..16413
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(16479..490)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg
       61 ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct
      121 ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg
      181 acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga
      241 gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacggcg
      301 caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg
      361 attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca
      421 acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc
      481 gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcaggc
      541 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc
      601 gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct
      661 actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca gatagcagct
      721 cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga tcctccgccg
      781 gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc aacgttgcag
      841 ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt ttgctcattt
      901 tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca gcgcctggac
      961 ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg cggcggcagt
     1021 gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct cgtacccggc
     1081 cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa
     1141 ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc
     1201 ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga agtcggctgc
     1261 cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc
     1321 gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt
     1381 tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg gatcggaatc
     1441 gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg ttgcgatggt
     1501 cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc gttccccttg
     1561 cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg caagctgttt
     1621 tactcaaata cacatcacct ttttagacgg cggcgctcgg tttcttcagc ggccaagctg
     1681 gccggccagg ccgccagctt ggcatcagac aaaccggcca ggatttcatg cagccgcacg
     1741 gttgagacgt gcgcgggcgg ctcgaacacg tacccggccg cgatcatctc cgcctcgatc
     1801 tcttcggtaa tgaaaaacgg ttcgtcctgg ccgtcctggt gcggtttcat gcttgttcct
     1861 cttggcgttc attctcggcg gccgccaggg cgtcggcctc ggtcaatgcg tcctcacgga
     1921 aggcaccgcg ccgcctggcc tcggtgggcg tcacttcctc gctgcgctca agtgcgcggt
     1981 acagggtcga gcgatgcacg ccaagcagtg cagccgcctc tttcacggtg cggccttcct
     2041 ggtcgatcag ctcgcgggcg tgcgcgatct gtgccggggt gagggtaggg cgggggccaa
     2101 acttcacgcc tcgggccttg gcggcctcgc gcccgctccg ggtgcggtcg atgattaggg
     2161 aacgctcgaa ctcggcaatg ccggcgaaca cggtcaacac catgcggccg gccggcgtgg
     2221 tggtgtcggc ccacggctct gccaggctac gcaggcccgc gccggcctcc tggatgcgct
     2281 cggcaatgtc cagtaggtcg cgggtgctgc gggccaggcg gtctagcctg gtcactgtca
     2341 caacgtcgcc agggcgtagg tggtcaagca tcctggccag ctccgggcgg tcgcgcctgg
     2401 tgccggtgat cttctcggaa aacagcttgg tgcagccggc cgcgtgcagt tcggcccgtt
     2461 ggttggtcaa gtcctggtcg tcggtgctga cgcgggcata gcccagcagg ccagcggcgg
     2521 cgctcttgtt catggcgtaa tgtctccggt tctagtcgca agtattctac tttatgcgac
     2581 taaaacacgc gacaagaaaa cgccaggaaa agggcagggc ggcagcctgt cgcgtaactt
     2641 aggacttgtg cgacatgtcg ttttcagaag acggctgcac tgaacgtcag aagccgactg
     2701 cactatagca gcggaggggt tggatcaaag tactttgatc ccgaggggaa ccctgtggtt
     2761 ggcatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgat
     2821 tattctaata aacgctcttt tctcttaggt ttacccgcca atatatcctg tcaaacactg
     2881 atagtttaaa ctgaaggcgg gaaacgacaa tctgatccaa gctcaagctg ctctagcatt
     2941 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     3001 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     3061 cccagtcacg acgttgtaaa acgacggcca gtgccaagct tcgacttgcc ttccgcacaa
     3121 tacatcattt cttcttagct ttttttcttc ttcttcgttc atacagtttt tttttgttta
     3181 tcagcttaca ttttcttgaa ccgtagcttt cgttttcttc tttttaactt tccattcgga
     3241 gtttttgtat cttgtttcat agtttgtccc aggattagaa tgattaggca tcgaaccttc
     3301 aagaatttga ttgaataaaa catcttcatt cttaagatat gaagataatc ttcaaaaggc
     3361 ccctgggaat ctgaaagaag agaagcaggc ccatttatat gggaaagaac aatagtattt
     3421 cttatatagg cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg cttagataag
     3481 aaaacgaagc tgagtttata tacagctaga gtcgaagtag tgattgggag accaacccag
     3541 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
     3601 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     3661 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
     3721 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
     3781 cgccctaaaa caaagttaaa catcatgggg gaagcggtga tcgccgaagt atcgactcaa
     3841 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
     3901 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
     3961 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
     4021 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
     4081 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
     4141 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
     4201 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
     4261 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
     4321 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     4381 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     4441 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     4501 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     4561 gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc tagctagaaa ttcgttcaag
     4621 ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga tgcactaagc acataattgc
     4681 tcacagccaa actatcaggt caagtctgct tttattattt ttaagcgtgc ataataagcc
     4741 ggtctcggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     4801 aaaaagtggc accgagtcgg tgcttttttt tgcaaaattt tccagatcga tttcttcttc
     4861 ctctgttctt cggcgttcaa tttctggggt tttctcttcg ttttctgtaa ctgaaaccta
     4921 aaatttgacc taaaaaaaat ctcaaataat atgattcagt ggttttgtac ttttcagtta
     4981 gttgagtttt gcagttccga tgagataaac caataagctt gcatgcctgc aggtcaacat
     5041 ggtggagcac gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca
     5101 aagggcaatt gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg
     5161 cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg
     5221 ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa
     5281 agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc
     5341 aaagcaagtg gattgatgtg ataacatggt ggagcacgac acacttgtct actccaaaaa
     5401 tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac aaagggtaat
     5461 atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg tgaagatagt
     5521 ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ccatcgttga
     5581 agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga
     5641 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
     5701 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
     5761 ttcatttcat ttggagagga cctcgacctc aacacaacat atacaaaaca aacgaatctc
     5821 aagcaatcaa gcattctact tctattgcag caatttaaat catttctttt aaagcaaaag
     5881 caattttctg aaaattttca ccatttacga acgatactcg agtaatatgc caaagaagaa
     5941 gaggaaggtt tcatcggaga ccggccctgt tgctgttgac cccaccctgc ggcggagaat
     6001 cgagccacac gagttcgagg tgttcttcga cccaagggag ctccgcaagg agacgtgcct
     6061 cctgtacgag atcaactggg gcggcaggca ctccatctgg aggcacacca gccaaaacac
     6121 caacaagcac gtggaggtca acttcatcga gaagttcacc accgagaggt acttctgccc
     6181 aaacacccgc tgctccatca cctggttcct gtcctggagc ccatgcggcg agtgctccag
     6241 ggccatcacc gagttcctca gccgctaccc acacgtcacc ctgttcatct acatcgccag
     6301 gctctaccac cacgccgacc caaggaacag gcagggcctc cgcgacctga tctccagcgg
     6361 cgtgaccatc caaatcatga ccgagcagga gtccggctac tgctggagga acttcgtcaa
     6421 ctactcccca agcaacgagg cccactggcc aaggtaccca cacctctggg tgcgcctcta
     6481 cgtgctcgag ctgtactgca tcatcctcgg cctgccacca tgcctcaaca tcctgaggcg
     6541 caagcaacca cagctgacct tcttcaccat cgccctccaa agctgccact accagaggct
     6601 cccaccacac atcctgtggg ctaccggcct caagtccggc agcgagacgc caggcacctc
     6661 cgagagcgct acgcctgaac ttaaggacaa gaagtactcg atcggcctcg ccatcgggac
     6721 gaactcagtt ggctgggccg tgatcaccga cgagtacaag gtgccctcta agaagttcaa
     6781 ggtcctgggg aacaccgacc gccattccat caagaagaac ctcatcggcg ctctcctgtt
     6841 cgacagcggg gagaccgctg aggctacgag gctcaagaga accgctaggc gccggtacac
     6901 gagaaggaag aacaggatct gctacctcca agagattttc tccaacgaga tggccaaggt
     6961 tgacgattca ttcttccacc gcctggagga gtctttcctc gtggaggagg ataagaagca
     7021 cgagcggcat cccatcttcg gcaacatcgt ggacgaggtt gcctaccacg agaagtaccc
     7081 tacgatctac catctgcgga agaagctcgt ggactccacc gataaggcgg acctcagact
     7141 gatctacctc gctctggccc acatgatcaa gttccgcggc catttcctga tcgaggggga
     7201 tctcaaccca gacaacagcg atgttgacaa gctgttcatc caactcgtgc agacctacaa
     7261 ccaactcttc gaggagaacc cgatcaacgc ctctggcgtg gacgcgaagg ctatcctgtc
     7321 cgcgaggctc tcgaagtcca ggaggctgga gaacctgatc gctcagctcc caggcgagaa
     7381 gaagaacggc ctgttcggga acctcatcgc tctcagcctg gggctcaccc cgaacttcaa
     7441 gtcgaacttc gatctcgctg aggacgccaa gctgcaactc tccaaggaca cctacgacga
     7501 tgacctcgat aacctcctgg cccagatcgg cgatcaatac gcggacctgt tcctcgctgc
     7561 caagaacctg tcggacgcca tcctcctgtc agatatcctc cgcgtgaaca ccgagatcac
     7621 gaaggctcca ctctctgcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac
     7681 cctcctgaag gcgctggtcc gccaacagct cccggagaag tacaaggaga ttttcttcga
     7741 tcagtcgaag aacggctacg ctgggtacat cgacggcggg gcctcacaag aggagttcta
     7801 caagttcatc aagccaatcc tggagaagat ggacggcacg gaggagctcc tggtgaagct
     7861 caacagggag gacctcctgc ggaagcagag aaccttcgat aacggcagca tcccccacca
     7921 aatccatctc ggggagctgc acgccatcct gagaaggcaa gaggacttct accctttcct
     7981 caaggataac cgggagaaga tcgagaagat cctgaccttc agaatcccat actacgtcgg
     8041 ccctctcgcg cgggggaact caagattcgc ttggatgacc cgcaagtctg aggagaccat
     8101 cacgccgtgg aacttcgagg aggtggtgga caagggcgct agcgctcagt cgttcatcga
     8161 gaggatgacc aacttcgaca agaacctgcc caacgagaag gtgctcccta agcactcgct
     8221 cctgtacgag tacttcaccg tctacaacga gctcacgaag gtgaagtacg tcaccgaggg
     8281 catgcgcaag ccagcgttcc tgtccgggga gcagaagaag gctatcgtgg acctcctgtt
     8341 caagaccaac cggaaggtca cggttaagca actcaaggag gactacttca agaagatcga
     8401 gtgcttcgat tcggtcgaga tcagcggcgt tgaggaccgc ttcaacgcca gcctcgggac
     8461 ctaccacgat ctcctgaaga tcatcaagga taaggacttc ctggacaacg aggagaacga
     8521 ggatatcctg gaggacatcg tgctgaccct cacgctgttc gaggacaggg agatgatcga
     8581 ggagcgcctg aagacgtacg cccatctctt cgatgacaag gtcatgaagc aactcaagcg
     8641 ccggagatac accggctggg ggaggctgtc ccgcaagctc atcaacggca tccgggacaa
     8701 gcagtccggg aagaccatcc tcgacttcct caagagcgat ggcttcgcca acaggaactt
     8761 catgcaactg atccacgatg acagcctcac cttcaaggag gatatccaaa aggctcaagt
     8821 gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cccccgcgat
     8881 caagaagggc atcctccaga ccgtgaaggt tgtggacgag ctcgtgaagg tcatgggccg
     8941 gcacaagcct gagaacatcg tcatcgagat ggccagagag aaccaaacca cgcagaaggg
     9001 gcaaaagaac tctagggagc gcatgaagcg catcgaggag ggcatcaagg agctggggtc
     9061 ccaaatcctc aaggagcacc cagtggagaa cacccaactg cagaacgaga agctctacct
     9121 gtactacctc cagaacggca gggatatgta cgtggaccaa gagctggata tcaaccgcct
     9181 cagcgattac gacgtcgatc atatcgttcc ccagtctttc ctgaaggatg actccatcga
     9241 caacaaggtc ctcaccaggt cggacaagaa ccgcggcaag tcagataacg ttccatctga
     9301 ggaggtcgtt aagaagatga agaactactg gaggcagctc ctgaacgcca agctgatcac
     9361 gcaaaggaag ttcgacaacc tcaccaaggc tgagagaggc gggctctcag agctggacaa
     9421 ggccggcttc atcaagcggc agctggtcga gaccagacaa atcacgaagc acgttgcgca
     9481 aatcctcgac tctcggatga acacgaagta cgatgagaac gacaagctga tcagggaggt
     9541 taaggtgatc accctgaagt ctaagctcgt ctccgacttc aggaaggatt tccagttcta
     9601 caaggttcgc gagatcaaca actaccacca tgcccatgac gcttacctca acgctgtggt
     9661 cggcaccgct ctgatcaaga agtacccaaa gctggagtcc gagttcgtgt acggggacta
     9721 caaggtttac gatgtgcgca agatgatcgc caagtcggag caagagatcg gcaaggctac
     9781 cgccaagtac ttcttctact caaacatcat gaacttcttc aagaccgaga tcacgctggc
     9841 caacggcgag atccggaaga gaccgctcat cgagaccaac ggcgagacgg gggagatcgt
     9901 gtgggacaag ggcagggatt tcgcgaccgt ccgcaaggtt ctctccatgc cccaggtgaa
     9961 catcgtcaag aagaccgagg tccaaacggg cgggttctca aaggagtcta tcctgcctaa
    10021 gcggaacagc gacaagctca tcgccagaaa gaaggactgg gacccaaaga agtacggcgg
    10081 gttcgacagc cctaccgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa
    10141 gtccaagaag ctcaagagcg tgaaggagct cctggggatc accatcatgg agaggtccag
    10201 cttcgagaag aacccaatcg acttcctgga ggccaagggc tacaaggagg tgaagaagga
    10261 cctgatcatc aagctcccga agtactctct cttcgagctg gagaacggca ggaagagaat
    10321 gctggcttcc gctggcgagc tccagaaggg gaacgagctc gcgctgccaa gcaagtacgt
    10381 gaacttcctc tacctggctt cccactacga gaagctcaag ggcagcccgg aggacaacga
    10441 gcaaaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca tcgagcaaat
    10501 ctccgagttc agcaagcgcg tgatcctcgc cgacgcgaac ctggataagg tcctctccgc
    10561 ctacaacaag caccgggaca agcccatcag agagcaagcg gagaacatca tccatctctt
    10621 caccctgacg aacctcggcg ctcctgctgc tttcaagtac ttcgacacca cgatcgatcg
    10681 gaagagatac acctccacga aggaggtcct ggacgcgacc ctcatccacc agtcgatcac
    10741 cggcctgtac gagacgagga tcgacctctc acaactcggc ggggataaga gacccgcagc
    10801 aaccaagaag gcagggcaag caaagaagaa gaagacgcgt gactccggcg gcagcaccaa
    10861 cctgtccgac atcatcgaga aggagacggg caagcaactc gtgatccagg agagcatcct
    10921 catgctgcca gaggaggtgg aggaggtcat cggcaacaag ccagagtccg acatcctggt
    10981 gcacaccgcc tacgacgagt ccaccgacga gaacgtcatg ctcctgacca gcgacgcccc
    11041 agagtacaag ccatgggccc tcgtcatcca ggacagcaac ggggagaaca agatcaagat
    11101 gctgtcgggg gggagcccaa agaagaagcg gaaggtgtag ctcagagctt tcgttcgtat
    11161 catcggtttc gacaacgttc gtcaagttca atgcatcagt ttcattgcgc acacaccaga
    11221 atcctactga gtttgagtat tatggcattg ggaaaactgt ttttcttgta ccatttgttg
    11281 tgcttgtaat ttactgtgtt ttttattcgg ttttcgctat cgaactgtga aatggaaatg
    11341 gatggagaag agttaatgaa tgatatggtc cttttgttca ttctcaaatt aatattattt
    11401 gttttttctc ttatttgttg tgtgttgaat ttgaaattat aagagatatg caaacatttt
    11461 gttttgagta aaaatgtgtc aaatcgtggc ctctaatgac cgaagttaat atgaggagta
    11521 aaacacttgt agttgtacca ttatgcttat tcactaggca acaaatatat tttcagacct
    11581 agaaaagctg caaatgttac tgaatacaag tatgtcctct tgtgttttag acatttatga
    11641 actttccttt atgtaatttt ccagaatcct tgtcagattc taatcattgc tttataatta
    11701 tagttatact catggatttg tagttgagta tgaaaatatt ttttaatgca ttttatgact
    11761 tgccaattga ttgacaacga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
    11821 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    11881 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    11941 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    12001 cgtattggct agagcagctt gccaacatgg tggagcacga cactctcgtc tactccaaga
    12061 atatcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa
    12121 tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag
    12181 tagaaaagga aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc
    12241 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
    12301 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat aacatggtgg
    12361 agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg
    12421 ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag
    12481 ctatctgtca cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc
    12541 attgcgataa aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg
    12601 gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
    12661 aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt
    12721 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca cgctgaaatc
    12781 accagtctct ctctacaaat ctatctctct cgagtctacc atgagcccag aacgacgccc
    12841 ggccgacatc cgccgtgcca ccgaggcgga catgccggcg gtctgcacca tcgtcaacca
    12901 ctacatcgag acaagcacgg tcaacttccg taccgagccg caggaaccgc aggagtggac
    12961 ggacgacctc gtccgtctgc gggagcgcta tccctggctc gtcgccgagg tggacggcga
    13021 ggtcgccggc atcgcctacg cgggcccctg gaaggcacgc aacgcctacg actggacggc
    13081 cgagtcgacc gtgtacgtct ccccccgcca ccagcggacg ggactgggct ccacgctcta
    13141 cacccacctg ctgaagtccc tggaggcaca gggcttcaag agcgtggtcg ctgtcatcgg
    13201 gctgcccaac gacccgagcg tgcgcatgca cgaggcgctc ggatatgccc cccgcggcat
    13261 gctgcgggcg gccggcttca agcacgggaa ctggcatgac gtgggtttct ggcagctgga
    13321 cttcagcctg ccggtaccgc cccgtccggt cctgcccgtc accgagattt gactcgagtt
    13381 tctccataat aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc
    13441 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
    13501 atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag atcccccgaa
    13561 ttaattcggc gttaattcag tacattaaaa acgtccgcaa tgtgttatta agttgtctaa
    13621 gcgtcaattt gtttacacca caatatatcc tgccaccagc cagccaacag ctccccgacc
    13681 ggcagctcgg cacaaaatca ccactcgata caggcagccc atcagtccgg gacggcgtca
    13741 gcgggagagc cgttgtaagg cggcagactt tgctcatgtt accgatgcta ttcggaagaa
    13801 cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag catgttgatt
    13861 gtaacgatga cagagcgttg ctgcctgtga tcaccgcggt ttcaaaatcg gctccgtcga
    13921 tactatgtta tacgccaact ttgaaaacaa ctttgaaaaa gctgttttct ggtatttaag
    13981 gttttagaat gcaaggaaca gtgaattgga gttcgtcttg ttataattag cttcttgggg
    14041 tatctttaaa tactgtagaa aagaggaagg aaataataaa tggctaaaat gagaatatca
    14101 ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct
    14161 cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac
    14221 agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg
    14281 gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat
    14341 ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc
    14401 cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata
    14461 tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg
    14521 aataacgatc tggccgatgt ggattgcgaa aactgggaag aagacactcc atttaaagat
    14581 ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc
    14641 cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt
    14701 gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg
    14761 atcagggagg atatcgggga agaacagtat gtcgagctat tttttgactt actggggatc
    14821 aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt ttagtaccta
    14881 gaatgcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
    14941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
    15001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
    15061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
    15121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
    15181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
    15241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
    15301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
    15361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
    15421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
    15481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
    15541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
    15601 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
    15661 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
    15721 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
    15781 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
    15841 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
    15901 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
    15961 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagggtg
    16021 ccttgatgtg ggcgccggcg gtcgagtggc gacggcgcgg cttgtccgcg ccctggtaga
    16081 ttgcctggcc gtaggccagc catttttgag cggccagcgg ccgcgatagg ccgacgcgaa
    16141 gcggcggggc gtagggagcg cagcgaccga agggtaggcg ctttttgcag ctcttcggct
    16201 gtgcgctggc cagacagtta tgcacaggcc aggcgggttt taagagtttt aataagtttt
    16261 aaagagtttt aggcggaaaa atcgcctttt ttctctttta tatcagtcac ttacatgtgt
    16321 gaccggttcc caatgtacgg ctttgggttc ccaatgtacg ggttccggtt cccaatgtac
    16381 ggctttgggt tcccaatgta cgtgctatcc acaggaaaca gaccttttcg acctttttcc
    16441 cctgctaggg caatttgccc tagcatctgc tccgtacatt aggaaccggc ggatgcttcg
    16501 ccctcgatca ggttgcggta gcgcatgact aggatcgggc cagcctgccc cgcctcctcc
    16561 ttcaaatcgt actccggcag gtcatttgac ccgatcagct tgcgcacggt gaaacagaac
    16621 ttcttgaact ctccggcgct gccactgcgt tcgtagatcg tcttgaacaa ccatctggct
    16681 tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa aacggccgat gccgggatcg
    16741 atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg
    16801 cggtacatcc aatcagctag ctcgatctcg atgtactccg gccgcccggt ttcgctcttt
    16861 acgatcttgt agcggctaat caaggcttca ccctcggata ccgtcaccag gcggccgttc
    16921 ttggccttct tcgtacgctg catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct
    16981 accaggtcgt ctttctgctt tccgccatcg gctcgccggc agaacttgag tacgtccgca
    17041 acgtgtggac ggaacacgcg gc
//