pH-STEME-1-esgRNA

LOCUS       pH-STEME-1-esgRNA       18514 bp DNA     circular SYN 26-AUG-2024
DEFINITION  Targeted simultaneous C-to-T and A-to-G in rice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18514)
  TITLE     Gao Lab Plasmids
REFERENCE   2  (bases 1 to 18514)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2024 from SnapGene Server 7.0.3
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..18514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(578..1207)
                     /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    2508..2532
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     2720..2742
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2734..2751
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2735..2751
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2818..3197
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     terminator      3305..3324
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        3337..5329
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             7269..11369
                     /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"
     RBS             11282..11290
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             11370..11417
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             11436..11684
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             11697..11717
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(12376..12392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12376..12392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12389..12411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12400..12416
                     /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(12424..12454)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12469..12490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12681..13357
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     13294..13314
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             13424..14449
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    14490..14664
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    14742..14766
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             15191..15985
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      16072..16660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     16561..16580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     16814..16831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    16846..16986
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(16912..16931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     17072..17094
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      17330..17524
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(17590..18514,1..149))
                     /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 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
       61 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
      121 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
      181 cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
      241 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
      301 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
      361 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
      421 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
      481 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
      541 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
      601 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
      661 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
      721 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
      781 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
      841 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
      901 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
      961 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
     1021 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
     1081 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
     1141 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
     1201 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
     1261 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
     1321 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
     1381 gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc
     1441 gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg
     1501 cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg
     1561 gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg
     1621 ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct
     1681 ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg
     1741 agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg
     1801 gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc
     1861 atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg
     1921 ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg
     1981 tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc
     2041 tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc
     2101 gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag
     2161 cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa
     2221 gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg
     2281 gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact
     2341 gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc
     2401 cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac
     2461 gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa
     2521 tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag
     2581 ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2641 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2701 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
     2761 agtaattcat ccaggtcacc aagttctagg attttcagaa ctgcaactta ttttatcaag
     2821 gaatctttaa acatacgaac agatcactta aagttcttct gaagcaactt aaagttatca
     2881 ggcatgcatg gatcttggag gaatcagatg tgcagtcagg gaccatagca caagacaggc
     2941 gtcttctact ggtgctacca gcaaatgctg gaagccggga acactgggta cgttggaaac
     3001 cacgtgatgt gaagaagtaa gataaactgt aggagaaaag catttcgtag tgggccatga
     3061 agcctttcag gacatgtatt gcagtatggg ccggcccatt acgcaattgg acgacaacaa
     3121 agactagtat tagtaccacc tcggctatcc acatagatca aagctgattt aaaagagttg
     3181 tgcagatgat ccgtggcgtg agaccctgca gggtctctgt ttaagagcta tgctggaaac
     3241 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     3301 gtgctttttt tgttttttat gtctaagctt gcatgcctgc agtgcagcgt gacccggtcg
     3361 tgcccctctc tagagataat gagcattgca tgtctaagtt ataaaaaatt accacatatt
     3421 ttttttgtca cacttgtttg aagtgcagtt tatctatctt tatacatata tttaaacttt
     3481 actctacgaa taatataatc tatagtacta caataatatc agtgttttag agaatcatat
     3541 aaatgaacag ttagacatgg tctaaaggac aattgagtat tttgacaaca ggactctaca
     3601 gttttatctt tttagtgtgc atgtgttctc cttttttttt gcaaatagct tcacctatat
     3661 aatacttcat ccattttatt agtacatcca tttagggttt agggttaatg gtttttatag
     3721 actaattttt ttagtacatc tattttattc tattttagcc tctaaattaa gaaaactaaa
     3781 actctatttt agttttttta tttaataatt tagatataaa atagaataaa ataaagtgac
     3841 taaaaattaa acaaataccc tttaagaaat taaaaaaact aaggaaacat ttttcttgtt
     3901 tcgagtagat aatgccagcc tgttaaacgc cgtcgacgag tctaacggac accaaccagc
     3961 gaaccagcag cgtcgcgtcg ggccaagcga agcagacggc acggcatctc tgtcgctgcc
     4021 tctggacccc tctcgagagt tccgctccac cgttggactt gctccgctgt cggcatccag
     4081 aaattgcgtg gcggagcggc agacgtgagc cggcacggca ggcggcctcc tcctcctctc
     4141 acggcaccgg cagctacggg ggattccttt cccaccgctc cttcgctttc ccttcctcgc
     4201 ccgccgtaat aaatagacac cccctccaca ccctctttcc ccaacctcgt gttgttcgga
     4261 gcgcacacac acacaaccag atctccccca aatccacccg tcggcacctc cgcttcaagg
     4321 tacgccgctc gtcctccccc cccccccctc tctaccttct ctagatcggc gttccggtcc
     4381 atggttaggg cccggtagtt ctacttctgt tcatgtttgt gttagatccg tgtttgtgtt
     4441 agatccgtgc tgctagcgtt cgtacacgga tgcgacctgt acgtcagaca cgttctgatt
     4501 gctaacttgc cagtgtttct ctttggggaa tcctgggatg gctctagccg ttccgcagac
     4561 gggatcgatt tcatgatttt ttttgtttcg ttgcataggg tttggtttgc ccttttcctt
     4621 tatttcaata tatgccgtgc acttgtttgt cgggtcatct tttcatgctt ttttttgtct
     4681 tggttgtgat gatgtggtct ggttgggcgg tcgttctaga tcggagtaga attctgtttc
     4741 aaactacctg gtggatttat taattttgga tctgtatgtg tgtgccatac atattcatag
     4801 ttacgaattg aagatgatgg atggaaatat cgatctagga taggtataca tgttgatgcg
     4861 ggttttactg atgcatatac agagatgctt tttgttcgct tggttgtgat gatgtggtgt
     4921 ggttgggcgg tcgttcattc gttctagatc ggagtagaat actgtttcaa actacctggt
     4981 gtatttatta attttggaac tgtatgtgtg tgtcatacat cttcatagtt acgagtttaa
     5041 gatggatgga aatatcgatc taggataggt atacatgttg atgtgggttt tactgatgca
     5101 tatacatgat ggcatatgca gcatctattc atatgctcta accttgagta cctatctatt
     5161 ataataaaca agtatgtttt ataattattt tgatcttgat atacttggat gatggcatat
     5221 gcagcagcta tatgtggatt tttttagccc tgccttcata cgctatttat ttgcttggta
     5281 ctgtttcttt tgtcgatgct caccctgttg tttggtgtta cttctgcagc cctaggccta
     5341 tggaggccag cccggctagc ggcccaaggc atctcatgga cccgcacatc ttcaccagca
     5401 acttcaacaa cggcatcggc aggcacaaga cctacttgtg ctacgaggtg gagaggctcg
     5461 acaacggaac ctccgtgaag atggaccaac acagggggtt cctccacaac caagccaaga
     5521 acctcctctg cggcttctac ggcaggcacg ccgagttgag gttcctcgac ttggtgccat
     5581 ccctccaact cgatccagcc caaatctacc gcgtgacctg gttcatctcc tggtccccat
     5641 gcttctcctg gggttgcgcc ggcgaggttc gggctttcct ccaagaaaac acccacgtcc
     5701 gcctccgcat tttcgccgcc aggatctatg attacgaccc tctctacaag gaggccctcc
     5761 agatgctgcg ggacgccggt gctcaggtga gtatcatgac ctacgacgag ttcaagcact
     5821 gctgggacac cttcgttgac caccagggct gcccattcca accatgggac ggtctggatg
     5881 aacacagcca agccttgtcc ggcaggctcc gggccatcct ccaaaaccag gggaactccg
     5941 ggagcgagac gccaggcacc tccgagtcgg ccaccccaga atcttccggc ggatcttctg
     6001 gaggatctag cggctccgag acaccaggaa catccgaatc cgctacacca gagtcttctg
     6061 gaggatctag cggaggatct tccgaggtgg agttctctca cgagtattgg atgaggcacg
     6121 ctcttacact tgctaagaga gcttgggacg aaagagaagt gccagttggc gccgttcttg
     6181 tgcataataa tagggtgatc ggcgagggtt ggaatagacc aattggaagg catgatccaa
     6241 cagctcacgc agagattatg gctctcagac aaggcggcct cgttatgcag aactacaggc
     6301 tcattgacgc tacactctac gtgacactcg aaccttgcgt tatgtgcgcc ggagctatga
     6361 ttcattctag gattggcagg gtcgtgtttg gagctaggga cgctaaaaca ggagccgccg
     6421 gatctcttat ggacgtgttg catcatccag gcatgaacca tagggtggag attacagagg
     6481 gcattcttgc agacgagtgc gctgctcttc tttccgattt cttcaggatg agaaggcagg
     6541 agattaaggc ccagaagaag gctcagtctt ctacagatag cggaggatct tccggaggat
     6601 ctagcggctc cgagacacca ggaacatccg aaagcgctac accagaatct agcggaggct
     6661 cttccggagg atcttctgaa gtggagttct cccacgagta ttggatgagg cacgctctta
     6721 cacttgctaa aagggctagg gacgaaaggg aagttccagt tggagctgtt ctcgtgctca
     6781 ataacagggt gattggcgag ggttggaata gagccattgg actccatgat ccaacagctc
     6841 acgcagagat tatggctctt agacaaggcg gcctcgttat gcagaattac agactcatcg
     6901 acgccacact ctacgttacc ttcgaacctt gcgttatgtg cgccggagct atgatccatt
     6961 ctaggattgg cagggtcgtg ttcggcgtta gaaacgctaa gacaggagct gcaggctctc
     7021 ttatggacgt tcttcattac ccaggcatga atcatagagt ggagatcaca gaaggcattc
     7081 ttgcagacga gtgcgcagct ctcctttgct atttcttcag gatgccgagg caagttttca
     7141 acgctcagaa gaaggcccag tcttctacag attccggcgg atcttctgga ggatctagcg
     7201 gctccgagac accaggaaca tccgaatccg ctacaccaga gtcttctgga ggatctagcg
     7261 gaggatctga caagaagtac tcgatcggcc tcgccattgg gactaactct gttggctggg
     7321 ccgtgatcac cgacgagtac aaggtgccct caaagaagtt caaggtcctg ggcaacaccg
     7381 atcggcattc catcaagaag aatctcattg gcgctctcct gttcgacagc ggcgagacgg
     7441 ctgaggctac gcggctcaag cgcaccgccc gcaggcggta cacgcgcagg aagaatcgca
     7501 tctgctacct gcaggagatt ttctccaacg agatggcgaa ggttgacgat tctttcttcc
     7561 acaggctgga ggagtcattc ctcgtggagg aggataagaa gcacgagcgg catccaatct
     7621 tcggcaacat tgtcgacgag gttgcctacc acgagaagta ccctacgatc taccatctgc
     7681 ggaagaagct cgtggactcc acagataagg cggacctccg cctgatctac ctcgctctgg
     7741 cccacatgat taagttcagg ggccatttcc tgatcgaggg ggatctcaac ccggacaata
     7801 gcgatgttga caagctgttc atccagctcg tgcagacgta caaccagctc ttcgaggaga
     7861 accccattaa tgcgtcaggc gtcgacgcga aggctatcct gtccgctagg ctctcgaagt
     7921 ctcggcgcct cgagaacctg atcgcccagc tgccgggcga gaagaagaac ggcctgttcg
     7981 ggaatctcat tgcgctcagc ctggggctca cgcccaactt caagtcgaat ttcgatctcg
     8041 ctgaggacgc caagctgcag ctctccaagg acacatacga cgatgacctg gataacctcc
     8101 tggcccagat cggcgatcag tacgcggacc tgttcctcgc tgccaagaat ctgtcggacg
     8161 ccatcctcct gtctgatatt ctcagggtga acaccgagat tacgaaggct ccgctctcag
     8221 cctccatgat caagcgctac gacgagcacc atcaggatct gaccctcctg aaggcgctgg
     8281 tcaggcagca gctccccgag aagtacaagg agatcttctt cgatcagtcg aagaacggct
     8341 acgctgggta cattgacggc ggggcctctc aggaggagtt ctacaagttc atcaagccga
     8401 ttctggagaa gatggacggc acggaggagc tgctggtgaa gctcaatcgc gaggacctcc
     8461 tgaggaagca gcggacattc gataacggca gcatcccaca ccagattcat ctcggggagc
     8521 tgcacgctat cctgaggagg caggaggact tctacccttt cctcaaggat aaccgcgaga
     8581 agatcgagaa gattctgact ttcaggatcc cgtactacgt cggcccactc gctaggggca
     8641 actcccgctt cgcttggatg acccgcaagt cagaggagac gatcacgccg tggaacttcg
     8701 aggaggtggt cgacaagggc gctagcgctc agtcgttcat cgagaggatg acgaatttcg
     8761 acaagaacct gccaaatgag aaggtgctcc ctaagcactc gctcctgtac gagtacttca
     8821 cagtctacaa cgagctgact aaggtgaagt atgtgaccga gggcatgagg aagccggctt
     8881 tcctgtctgg ggagcagaag aaggccatcg tggacctcct gttcaagacc aaccggaagg
     8941 tcacggttaa gcagctcaag gaggactact tcaagaagat tgagtgcttc gattcggtcg
     9001 agatctctgg cgttgaggac cgcttcaacg cctccctggg gacctaccac gatctcctga
     9061 agatcattaa ggataaggac ttcctggaca acgaggagaa tgaggatatc ctcgaggaca
     9121 ttgtgctgac actcactctg ttcgaggacc gggagatgat cgaggagcgc ctgaagactt
     9181 acgcccatct cttcgatgac aaggtcatga agcagctcaa gaggaggagg tacaccggct
     9241 gggggaggct gagcaggaag ctcatcaacg gcattcggga caagcagtcc gggaagacga
     9301 tcctcgactt cctgaagagc gatggcttcg cgaaccgcaa tttcatgcag ctgattcacg
     9361 atgacagcct cacattcaag gaggatatcc agaaggctca ggtgagcggc cagggggact
     9421 cgctgcacga gcatatcgcg aacctcgctg gctcgccagc tatcaagaag gggattctgc
     9481 agaccgtgaa ggttgtggac gagctggtga aggtcatggg caggcacaag cctgagaaca
     9541 tcgtcattga gatggcccgg gagaatcaga ccacgcagaa gggccagaag aactcacgcg
     9601 agaggatgaa gaggatcgag gagggcatta aggagctggg gtcccagatc ctcaaggagc
     9661 acccggtgga gaacacgcag ctgcagaatg agaagctcta cctgtactac ctccagaatg
     9721 gccgcgatat gtatgtggac caggagctgg atattaacag gctcagcgat tacgacgtcg
     9781 atcatatcgt tccacagtca ttcctgaagg atgactccat tgacaacaag gtcctcacca
     9841 ggtcggacaa gaaccggggc aagtctgata atgttccttc agaggaggtc gttaagaaga
     9901 tgaagaacta ctggcgccag ctcctgaatg ccaagctgat cacgcagcgg aagttcgata
     9961 acctcacaaa ggctgagagg ggcgggctct ctgagctgga caaggcgggc ttcatcaaga
    10021 ggcagctggt cgagacacgg cagatcacta agcacgttgc gcagattctc gactcacgga
    10081 tgaacactaa gtacgatgag aatgacaagc tgatccgcga ggtgaaggtc atcaccctga
    10141 agtcaaagct cgtctccgac ttcaggaagg atttccagtt ctacaaggtt cgggagatca
    10201 acaattacca ccatgcccat gacgcgtacc tgaacgcggt ggtcggcaca gctctgatca
    10261 agaagtaccc aaagctcgag agcgagttcg tgtacgggga ctacaaggtt tacgatgtga
    10321 ggaagatgat cgccaagtcg gagcaggaga ttggcaaggc taccgccaag tacttcttct
    10381 actctaacat tatgaatttc ttcaagacag agatcactct ggccaatggc gagatccgga
    10441 agcgccccct catcgagacg aacggcgaga cgggggagat cgtgtgggac aagggcaggg
    10501 atttcgcgac cgtcaggaag gttctctcca tgccacaagt gaatatcgtc aagaagacag
    10561 aggtccagac tggcgggttc tctaaggagt caattctgcc taagcggaac agcgacaagc
    10621 tcatcgcccg caagaaggac tgggatccga agaagtacgg cgggttcgac agccccactg
    10681 tggcctactc ggtcctggtt gtggcgaagg ttgagaaggg caagtccaag aagctcaaga
    10741 gcgtgaagga gctgctgggg atcacgatta tggagcgctc cagcttcgag aagaacccga
    10801 tcgatttcct ggaggcgaag ggctacaagg aggtgaagaa ggacctgatc attaagctcc
    10861 ccaagtactc actcttcgag ctggagaacg gcaggaagcg gatgctggct tccgctggcg
    10921 agctgcagaa ggggaacgag ctggctctgc cgtccaagta tgtgaacttc ctctacctgg
    10981 cctcccacta cgagaagctc aagggcagcc ccgaggacaa cgagcagaag cagctgttcg
    11041 tcgagcagca caagcattac ctcgacgaga tcattgagca gatttccgag ttctccaagc
    11101 gcgtgatcct ggccgacgcg aatctggata aggtcctctc cgcgtacaac aagcaccgcg
    11161 acaagccaat cagggagcag gctgagaata tcattcatct cttcaccctg acgaacctcg
    11221 gcgcccctgc tgctttcaag tacttcgaca caactatcga tcgcaagagg tacacaagca
    11281 ctaaggaggt cctggacgcg accctcatcc accagtcgat taccggcctc tacgagacgc
    11341 gcatcgacct gtctcagctc gggggcgaca agcggccagc ggcgacgaag aaggcggggc
    11401 aggcgaagaa gaagaaggaa ttctccggcg gcagcaccaa cctgtccgac atcatcgaga
    11461 aggagacggg caagcaactc gtgatccagg agagcatcct catgctgcca gaggaggtgg
    11521 aggaggtcat cggcaacaag ccagagtccg acatcctggt gcacaccgcc tacgacgagt
    11581 ccaccgacga gaacgtcatg ctcctgacca gcgacgcccc agagtacaag ccatgggccc
    11641 tcgtcatcca ggacagcaac ggggagaaca agatcaagat gctgtcgggg gggagcccaa
    11701 agaagaagcg gaaggtgtag gagctcagag ctttcgttcg tatcatcggt ttcgacaacg
    11761 ttcgtcaagt tcaatgcatc agtttcattg cgcacacacc agaatcctac tgagtttgag
    11821 tattatggca ttgggaaaac tgtttttctt gtaccatttg ttgtgcttgt aatttactgt
    11881 gttttttatt cggttttcgc tatcgaactg tgaaatggaa atggatggag aagagttaat
    11941 gaatgatatg gtccttttgt tcattctcaa attaatatta tttgtttttt ctcttatttg
    12001 ttgtgtgttg aatttgaaat tataagagat atgcaaacat tttgttttga gtaaaaatgt
    12061 gtcaaatcgt ggcctctaat gaccgaagtt aatatgagga gtaaaacact tgtagttgta
    12121 ccattatgct tattcactag gcaacaaata tattttcaga cctagaaaag ctgcaaatgt
    12181 tactgaatac aagtatgtcc tcttgtgttt tagacattta tgaactttcc tttatgtaat
    12241 tttccagaat ccttgtcaga ttctaatcat tgctttataa ttatagttat actcatggat
    12301 ttgtagttga gtatgaaaat attttttaat gcattttatg acttgccaat tgattgacaa
    12361 cgaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    12421 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    12481 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    12541 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctagagcag
    12601 cttgccaaca tggtggagca cgacactctc gtctactcca agaatatcaa agatacagtc
    12661 tcagaagacc aaagggctat tgagactttt caacaaaggg taatatcggg aaacctcctc
    12721 ggattccatt gcccagctat ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc
    12781 acctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc ctctgccgac
    12841 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
    12901 accacgtctt caaagcaagt ggattgatgt gaacatggtg gagcacgaca ctctcgtcta
    12961 ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga cttttcaaca
    13021 aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcaa
    13081 aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata aaggaaaggc
    13141 tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
    13201 catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat
    13261 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
    13321 ataaggaagt tcatttcatt tggagaggac acgctgaaat caccagtctc tctctacaaa
    13381 tctatctctc tcgagctttc gcagatccgg ggggcaatga gatatgaaaa agcctgaact
    13441 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
    13501 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
    13561 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
    13621 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt ttagcgagag
    13681 cctgacctat tgcatctccc gccgttcaca gggtgtcacg ttgcaagacc tgcctgaaac
    13741 cgaactgccc gctgttctac aaccggtcgc ggaggctatg gatgcgatcg ctgcggccga
    13801 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
    13861 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
    13921 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
    13981 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
    14041 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
    14101 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
    14161 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccacgactcc gggcgtatat
    14221 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
    14281 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
    14341 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc
    14401 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aagaaataga gtagatgccg
    14461 accgggatct gtcgatcgac aagctcgagt ttctccataa taatgtgtga gtagttccca
    14521 gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct
    14581 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
    14641 aaatccagta ctaaaatcca gatcccccga attaattcgg cgttaattca gtacattaaa
    14701 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
    14761 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
    14821 acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag gcggcagact
    14881 ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg tttgaaacac
    14941 ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt gctgcctgtg
    15001 atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac tttgaaaaca
    15061 actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac agtgaattgg
    15121 agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga aaagaggaag
    15181 gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga tcgaaaaata
    15241 ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc tggtgggaga
    15301 aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca cctatgatgt
    15361 ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc caaaggtcct
    15421 gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg atggcgtcct
    15481 ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc tgtatgcgga
    15541 gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga atagcttaga
    15601 cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg tggattgcga
    15661 aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt ttttaaagac
    15721 ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca gcaacatctt
    15781 tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca gggcggacaa
    15841 gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg aagaacagta
    15901 tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa taaaatatta
    15961 tattttactg gatgaattgt tttagtacct agaatgcatg accaaaatcc cttaacgtga
    16021 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    16081 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    16141 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    16201 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    16261 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    16321 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    16381 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    16441 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    16501 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    16561 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    16621 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    16681 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    16741 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    16801 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
    16861 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
    16921 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
    16981 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
    17041 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
    17101 ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc ggtcgagtgg
    17161 cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag ccatttttga
    17221 gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc gcagcgaccg
    17281 aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt atgcacaggc
    17341 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt
    17401 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt
    17461 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc
    17521 cacaggaaac agaccttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg
    17581 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac
    17641 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga
    17701 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg
    17761 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag
    17821 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag
    17881 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc
    17941 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc
    18001 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac
    18061 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc
    18121 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt
    18181 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag
    18241 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg
    18301 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg
    18361 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg
    18421 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg gcgcaccggc
    18481 tgccggcggt tgccgggatt ctttgcggat tcga
//