pGEL031

LOCUS       pGEL031         15756 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  STU2.0 SpyCas9 tRNA system for plant genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    pGEL031
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15756)
  AUTHORS   Tang X, Ren Q, Yang L, Bao Y, Zhong Z, He Y, Liu S, Qi C, Liu B, 
            Wang Y, Sretenovic S, Zhang Y, Zheng X, Zhang T, Qi Y, Zhang Y
  TITLE     Single transcript unit CRISPR 2.0 systems for robust Cas9 and Cas12a
            mediated plant genome editing.
  JOURNAL   Plant Biotechnol J. 2019 Jul;17(7):1431-1445. doi: 
            10.1111/pbi.13068. Epub 2019 Jan 17.
  PUBMED    30582653
REFERENCE   2  (bases 1 to 15756)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15756
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        8..2006
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             2031..6134
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6147..6167
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6311..6616)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(6413..6432)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        6943..7018
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    7372..7396
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8697..9326
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9755..10828
                     /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"
     rep_origin      10894..11088
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11324..11346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    11422..11562
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11477..11496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(11577..11594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11748..12336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11828..11847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12423..13217)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    13642..13666
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    13744..13918
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(15008..15684)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(15051..15071)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 ggcgcgcctg cagtgcagcg tgacccggtc gtgcccctct cttgagataa ggtgagcatt
       61 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca
      121 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta
      181 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag
      241 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt
      301 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat
      361 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta
      421 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata
      481 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga
      541 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa
      601 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag
      661 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc
      721 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg
      781 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc
      841 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc
      901 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagaactccc
      961 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc cccccccccc
     1021 ntctctacct tctcaagatc ggcgttccgg tccatggtta gggcccggta gttctacttc
     1081 tgttcatgtt tgtgttagat ccgtgtttgt gttagatccg tgctactagc gttcgtacac
     1141 ggatgcgacc tgtacgtcag acacgttctg attgctaact tgccagtgtt tctctttggg
     1201 gaatcctggg atggctctag ccgttccgca gacgggatcg atttcatgat tttttttgtt
     1261 tcgttgcata gggtttggtt tgcccttttc ctttatttca atatatgccg tgcacttgtt
     1321 tgtcgggtca tcttttcatg cttttttttg tcttggttgt gatgatgtgg tctggttggg
     1381 cggtcgttca agatcggagt agaattaatt ctgtttcaaa ctacctggtg gatttattaa
     1441 ttttggatct gtatgtgtgt gccatacata ttcatagtta cgaattgaag atgatggatg
     1501 gaaatatcga tctaggatag gtatacatgt tgatgcgggt tttactgatg catatacaga
     1561 gatgcttttt gttcgcttgg ttgtgatgat gtggtgtggt tgggcggtcg ttcattcgtt
     1621 caagatcgga gtagaatact gtttcaaact acctggtgta tttattaatt ttggaactgt
     1681 atgtgtgtgt catacatctt catagttacg agtttaagat ggatggaaat atcgatctag
     1741 gataggtata catgttgatg tgggttttac tgatgcatat acatgatggc atatgcagca
     1801 tctattcata tgctctaacc ttgagtacct atctattata ataaacaagt atgttttata
     1861 attattttga tcttgatata cttggatgat ggcatatgca gcagctatat gtggattttt
     1921 ttagccctgc cttcatacgc tatttatttg cttggtactg tttcttttgt cgatgctcac
     1981 cctgttgttt ggtgttactt ctgcagcctg caggtagatc gctcgtcgac atggataaga
     2041 agtactctat cggactcgat atcggaacta actctgtggg atgggctgtg atcaccgatg
     2101 agtacaaggt gccatctaag aagttcaagg ttctcggaaa caccgatagg cactctatca
     2161 agaaaaacct tatcggtgct ctcctcttcg attctggtga aactgctgag gctaccagac
     2221 tcaagagaac cgctagaaga aggtacacca gaagaaagaa caggatctgc tacctccaag
     2281 agatcttctc taacgagatg gctaaagtgg atgattcatt cttccacagg ctcgaagagt
     2341 cattcctcgt ggaagaagat aagaagcacg agaggcaccc tatcttcgga aacatcgttg
     2401 atgaggtggc ataccacgag aagtacccta ctatctacca cctcagaaag aagctcgttg
     2461 attctactga taaggctgat ctcaggctca tctacctcgc tctcgctcac atgatcaagt
     2521 tcagaggaca cttcctcatc gagggtgatc tcaaccctga taactctgat gtggataagt
     2581 tgttcatcca gctcgtgcag acctacaacc agcttttcga agagaaccct atcaacgctt
     2641 caggtgtgga tgctaaggct atcctctctg ctaggctctc taagtcaaga aggcttgaga
     2701 acctcattgc tcagctccct ggtgagaaga agaacggact tttcggaaac ttgatcgctc
     2761 tctctctcgg actcacccct aacttcaagt ctaacttcga tctcgctgag gatgcaaagc
     2821 tccagctctc aaaggatacc tacgatgatg atctcgataa cctcctcgct cagatcggag
     2881 atcagtacgc tgatttgttc ctcgctgcta agaacctctc tgatgctatc ctcctcagtg
     2941 atatcctcag agtgaacacc gagatcacca aggctccact ctcagcttct atgatcaaga
     3001 gatacgatga gcaccaccag gatctcacac ttctcaaggc tcttgttaga cagcagctcc
     3061 cagagaagta caaagagatt ttcttcgatc agtctaagaa cggatacgct ggttacatcg
     3121 atggtggtgc atctcaagaa gagttctaca agttcatcaa gcctatcctc gagaagatgg
     3181 atggaaccga ggaactcctc gtgaagctca atagagagga tcttctcaga aagcagagga
     3241 ccttcgataa cggatctatc cctcatcaga tccacctcgg agagttgcac gctatcctta
     3301 gaaggcaaga ggatttctac ccattcctca aggataacag ggaaaagatt gagaagattc
     3361 tcaccttcag aatcccttac tacgtgggac ctctcgctag aggaaactca agattcgctt
     3421 ggatgaccag aaagtctgag gaaaccatca ccccttggaa cttcgaagag gtggtggata
     3481 agggtgctag tgctcagtct ttcatcgaga ggatgaccaa cttcgataag aaccttccaa
     3541 acgagaaggt gctccctaag cactctttgc tctacgagta cttcaccgtg tacaacgagt
     3601 tgaccaaggt taagtacgtg accgagggaa tgaggaagcc tgcttttttg tcaggtgagc
     3661 aaaagaaggc tatcgttgat ctcttgttca agaccaacag aaaggtgacc gtgaagcagc
     3721 tcaaagagga ttacttcaag aaaatcgagt gcttcgattc agttgagatt tctggtgttg
     3781 aggataggtt caacgcatct ctcggaacct accacgatct cctcaagatc attaaggata
     3841 aggatttctt ggataacgag gaaaacgagg atatcttgga ggatatcgtt cttaccctca
     3901 ccctctttga agatagagag atgattgaag aaaggctcaa gacctacgct catctcttcg
     3961 atgataaggt gatgaagcag ttgaagagaa gaagatacac tggttgggga aggctctcaa
     4021 gaaagctcat taacggaatc agggataagc agtctggaaa gacaatcctt gatttcctca
     4081 agtctgatgg attcgctaac agaaacttca tgcagctcat ccacgatgat tctctcacct
     4141 ttaaagagga tatccagaag gctcaggttt caggacaggg tgatagtctc catgagcata
     4201 tcgctaacct cgctggatct cctgcaatca agaagggaat cctccagact gtgaaggttg
     4261 tggatgagtt ggtgaaggtg atgggaaggc ataagcctga gaacatcgtg atcgaaatgg
     4321 ctagagagaa ccagaccact cagaagggac agaagaactc tagggaaagg atgaagagga
     4381 tcgaggaagg tatcaaagag cttggatctc agatcctcaa agagcaccct gttgagaaca
     4441 ctcagctcca gaatgagaag ctctacctct actacctcca gaacggaagg gatatgtatg
     4501 tggatcaaga gttggatatc aacaggctct ctgattacga tgttgatcat atcgtgccac
     4561 agtcattctt gaaggatgat tctatcgata acaaggtgct caccaggtct gataagaaca
     4621 ggggtaagag tgataacgtg ccaagtgaag aggttgtgaa gaaaatgaag aactattgga
     4681 ggcagctcct caacgctaag ctcatcactc agagaaagtt cgataacttg actaaggctg
     4741 agaggggagg actctctgaa ttggataagg caggattcat caagaggcag cttgtggaaa
     4801 ccaggcagat cactaagcac gttgcacaga tcctcgattc taggatgaac accaagtacg
     4861 atgagaacga taagttgatc agggaagtga aggttatcac cctcaagtca aagctcgtgt
     4921 ctgatttcag aaaggatttc caattctaca aggtgaggga aatcaacaac taccaccacg
     4981 ctcacgatgc ttaccttaac gctgttgttg gaaccgctct catcaagaag tatcctaagc
     5041 tcgagtcaga gttcgtgtac ggtgattaca aggtgtacga tgtgaggaag atgatcgcta
     5101 agtctgagca agagatcgga aaggctaccg ctaagtattt cttctactct aacatcatga
     5161 atttcttcaa gaccgagatt accctcgcta acggtgagat cagaaagagg ccactcatcg
     5221 agacaaacgg tgaaacaggt gagatcgtgt gggataaggg aagggatttc gctaccgtta
     5281 gaaaggtgct ctctatgcca caggtgaaca tcgttaagaa aaccgaggtg cagaccggtg
     5341 gattctctaa agagtctatc ctccctaaga ggaactctga taagctcatt gctaggaaga
     5401 aggattggga ccctaagaaa tacggtggtt tcgattctcc taccgtggct tactctgttc
     5461 tcgttgtggc taaggttgag aagggaaaga gtaagaagct caagtctgtt aaggaacttc
     5521 tcggaatcac tatcatggaa aggtcatctt tcgagaagaa cccaatcgat ttcctcgagg
     5581 ctaagggata caaagaggtt aagaaggatc tcatcatcaa gctcccaaag tactcactct
     5641 tcgaactcga gaacggtaga aagaggatgc tcgcttctgc tggtgagctt caaaagggaa
     5701 acgagcttgc tctcccatct aagtacgtta actttcttta cctcgcttct cactacgaga
     5761 agttgaaggg atctccagaa gataacgagc agaagcaact tttcgttgag cagcacaagc
     5821 actacttgga tgagatcatc gagcagatct ctgagttctc taaaagggtg atcctcgctg
     5881 atgcaaacct cgataaggtg ttgtctgctt acaacaagca cagagataag cctatcaggg
     5941 aacaggcaga gaacatcatc catctcttca cccttaccaa cctcggtgct cctgctgctt
     6001 tcaagtactt cgatacaacc atcgatagga agagatacac ctctaccaaa gaagtgctcg
     6061 atgctaccct catccatcag tctatcactg gactctacga gactaggatc gatctctcac
     6121 agctcggtgg tgattcaagg gctgatccta agaagaagag gaaggtttga ggatctaaaa
     6181 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaca aagcaccagt
     6241 ggtctagtgg tagaatagta ccctgccacg gtacagaccc gggttcgatt cccggctggt
     6301 gcaggagacc ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc
     6361 ggtggctgag atcagccact tcttccccga taacggaaac cggcacactg gccatatcgg
     6421 tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacgggaga
     6481 ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt
     6541 caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt
     6601 aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag caaaagagcc gttcatttca
     6661 ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc
     6721 agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt gacatcatat
     6781 atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag
     6841 catacctctt tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa
     6901 atcagcgcgc aaatacgcat actgttatct ggcttggtct cagttttaga gctagaaata
     6961 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgcaa
     7021 caaagcacca gtggtctagt ggtagaatag taccctgcca cggtacagac ccgggttcga
     7081 ttcccggctg gtgcaggatc catatgaaga tgaagatgaa atatttggtg tgtcaaataa
     7141 aaagcttgtg tgcttaagtt tgtgtttttt tcttggcttg ttgtgttatg aatttgtggc
     7201 tttttctaat attaaatgaa tgtaagatca cattataatg aataaacaaa tgtttctata
     7261 atccattgtg aatgttttgt tggatctctt ctgcagcata taactactgt atgtgctatg
     7321 gtatggacta tggaatatga ttaaagataa ggagctcaaa ctatcagtgt ttgacaggat
     7381 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
     7441 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc
     7501 tcgggatcaa agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg
     7561 ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc
     7621 tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa
     7681 tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct
     7741 gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca
     7801 cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc
     7861 ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct
     7921 agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc
     7981 cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg
     8041 catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg
     8101 cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac
     8161 cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt
     8221 gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg
     8281 cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt
     8341 gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa
     8401 ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg
     8461 atcgcggccg ggtacgtgtt cgagccgccc gcgcacggct caaccgtgcg gctgcatgaa
     8521 atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa
     8581 gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat
     8641 gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga
     8701 aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc
     8761 tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg
     8821 gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg
     8881 accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg
     8941 acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc
     9001 tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg
     9061 ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg
     9121 cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc
     9181 ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca
     9241 caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg
     9301 ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac
     9361 aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag
     9421 cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac
     9481 caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata
     9541 catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg
     9601 ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca
     9661 tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgcctgc cggccctgca
     9721 atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc
     9781 ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc
     9841 cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc
     9901 tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa
     9961 gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac
    10021 ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg
    10081 agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc
    10141 ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac
    10201 cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc
    10261 acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga
    10321 cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa
    10381 ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa
    10441 gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat
    10501 gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt
    10561 tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa
    10621 ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt
    10681 caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga
    10741 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
    10801 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
    10861 agcaggggaa aaaggtcgaa aaggcctctt tcctgtggat agcacgtaca ttgggaaccc
    10921 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
    10981 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
    11041 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
    11101 gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc
    11161 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
    11221 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
    11281 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
    11341 tcccggaaac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcgcgtcagc
    11401 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
    11461 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
    11521 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
    11581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    11641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    11701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    11761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    11821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    11881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    11941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    12001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    12061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    12121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    12181 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    12241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    12301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    12361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcattctagg
    12421 tactaaaaca attcatccag taaaatataa tattttattt tctcccaatc aggcttgatc
    12481 cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc cctgatcgac
    12541 cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc aagatcaata
    12601 aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa
    12661 aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc gcgcggatct
    12721 ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc gttattcagt
    12781 aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca atccgatatg
    12841 tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt ttcagggctt
    12901 tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat gagcagattg
    12961 ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt tccttccagc
    13021 catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata ccggctgtcc
    13081 gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt agcaggagac
    13141 attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa ttccggtgat
    13201 attctcattt tagccattta ttatttcctt cctcttttct acagtattta aagatacccc
    13261 aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc taaaacctta
    13321 aataccagaa aacagctttt tcaaagttgt tttcaaagtt ggcgtataac atagtatcga
    13381 cggagccgat tttgaaaccg cggtgatcac aggcagcaac gctctgtcat cgttacaatc
    13441 aacatgctac cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct
    13501 tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac
    13561 gccgtcccgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
    13621 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag
    13681 acaacttaat aacacattgc ggacgttttt aatgtactga attaacgccg aattaattcg
    13741 ggggatctgg attttagtac tggattttgg ttttaggaat tagaaatttt attgatagaa
    13801 gtattttaca aatacaaata catactaagg gtttcttata tgctcaacac atgagcgaaa
    13861 ccctatagga accctaattc ccttatctgg gaactactca cacattatta tggagaaaat
    13921 agagcttgtc gatcgttaat taatcacctc caagatcccg gacgagtgct ggggcgtcgg
    13981 tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg
    14041 gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc
    14101 tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga
    14161 ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc
    14221 tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg
    14281 gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg
    14341 cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact
    14401 tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg
    14461 acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg
    14521 aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc
    14581 gtctggctaa gatcggccgc agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca
    14641 gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg
    14701 caataggtca ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg
    14761 gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt
    14821 acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc
    14881 tccgagagct gcatcaggtc ggacacgctg tcgaactttt cgatcagaaa cttctcgaca
    14941 gacgtcgcgg tgagttcagg ctttttcatc ctaggagaga tagatttgta gagagagact
    15001 ggtgatttca gcgtgtcctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt
    15061 gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt ggagatatca catcaatcca
    15121 cttgctttga agacgtggtt ggaacgtctt ctttttccac gatgctcctc gtgggtgggg
    15181 gtccatcttt gggaccactg tcggcagagg catcttgaac gatagccttt cctttatcgc
    15241 aatgatggca tttgtaggtg ccaccttcct tttctactgt ccttttgatg aagtgacaga
    15301 tagctgggca atggaatccg aggaggtttc ccgatattac cctttgttga aaagtctcaa
    15361 tagccctttg gtcttctgag actgtatctt tgatattctt ggagtagacg agagtgtcgt
    15421 gctccaccat gttcacatca atccacttgc tttgaagacg tggttggaac gtcttctttt
    15481 tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc agaggcatct
    15541 tgaacgatag cctttccttt atcgcaatga tggcatttgt aggtgccacc ttccttttct
    15601 actgtccttt tgatgaagtg acagatagct gggcaatgga atccgaggag gtttcccgat
    15661 attacccttt gttgaaaagt ctcaatagcc ctttggtctt ctgagactgt atctttgata
    15721 ttcttggagt agacgagagt gtcgtgctcc accatg
//