pBI121①植物表达质粒

基本信息

启动子:	CaMV 35S promoter
复制子:	oriV
终止子:	NOS
质粒分类:	植物系列，蛋白过表达载体
质粒大小:	14758bp
原核抗性:	卡那霉素Kan
筛选标记:	遗传霉素G418/新霉素Neo
克隆菌株:	大肠杆菌DH5α
培养条件:	37℃，有氧 LB
5'测序引物:	35S:GACGCACAATCCCACTATCC
3'测序引物:	根据序列设计引物

质粒简介

     pBI121载体是一个双元植物农杆菌表达载体，能够高效转染植物。该载体来源于pB221和Bin19载体。载体上含有从ColE1来源的ori复制元件，从CaMV中来源的pROK1元件，大肠杆菌新霉素磷酸转移酶II基因（Neomycin phosphotransferase II Gene, NptII），新霉素抗性基因(Neomycin resistant gene, Neo), 敏感基因Beta葡糖苷酸酶(glucuronidase)，Ter 农杆菌Ti 质粒胭脂氨酸合成酶原件等。pBI121载体是卡那抗性质粒。

质粒图谱

质粒序列

LOCUS       Exported               14758 bp ds-DNA     circular SYN 13-SEP-2016
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14758)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14758
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(145..516)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /note="traJ"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            549..658
                     /note="incP origin of transfer"
     CDS             complement(1339..1989)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /note="TetR"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2454..2478
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        2602..2781
                     /note="NOS promoter"
                     /note="nopaline synthase promoter"
     promoter        2634..2817
                     /function="NOS promoter"
                     /note="NOS promoter"
                     /note="nopaline synthase promoter"
     CDS             2838..3632
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      4022..4274
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4823..4844
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4859..4889
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4913
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     primer_bind     4921..4937
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5461..5806
                     /note="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5845..7656
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /note="GUS"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     terminator      7727..7979
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7988..8004)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8622..8646
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(10242..11390)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /note="trfA"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(11689..12483)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  12675..13442
                     /mobile_element_type="insertion sequence:IS1"
                     /note="IS1"
                     /note="prokaryotic transposable element"
     rep_origin      14065..18
                     /note="oriV"
                     /note="incP origin of replication"
ORIGIN
        1 tgagcgtcgc aaaggcgctc ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc
       61 gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc
      121 cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca cgcccatcat
      181 gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga ggatcgtggc
      241 atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca ggccgcccag
      301 gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc
      361 cgtgattttg tagccctggc cgacggccag caggtaggcc gacaggctca tgccggccgc
      421 cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct tgataggtgg
      481 gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc
      541 ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata
      601 agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg
      661 ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata
      721 tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta
      781 tgcagcggaa aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1021 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1081 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1141 cagtgagcga ggaagcggaa gagcgccaga aggccgccag agaggccgag cgcggccgtg
     1201 aggcttggac gctagggcag ggcatgaaaa agcccgtagc gggctgctac gggcgtctga
     1261 cgcggtggaa agggggaggg gatgttgtct acatggctct gctgtagtga gtgggttgcg
     1321 ctccggcagc ggtcctgatc aatcgtcacc ctttctcggt ccttcaacgt tcctgacaac
     1381 gagcctcctt ttcgccaatc catcgacaat caccgcgagt ccctgctcga acgctgcgtc
     1441 cggaccggct tcgtcgaagg cgtctatcgc ggcccgcaac agcggcgaga gcggagcctg
     1501 ttcaacggtg ccgccgcgct cgccggcatc gctgtcgccg gcctgctcct caagcacggc
     1561 cccaacagtg aagtagctga ttgtcatcag cgcattgacg gcgtccccgg ccgaaaaacc
     1621 cgcctcgcag aggaagcgaa gctgcgcgtc ggccgtttcc atctgcggtg cgcccggtcg
     1681 cgtgccggca tggatgcgcg cgccatcgcg gtaggcgagc agcgcctgcc tgaagctgcg
     1741 ggcattcccg atcagaaatg agcgccagtc gtcgtcggct ctcggcaccg aatgcgtatg
     1801 attctccgcc agcatggctt cggccagtgc gtcgagcagc gcccgcttgt tcctgaagtg
     1861 ccagtaaagc gccggctgct gaacccccaa ccgttccgcc agtttgcgtg tcgtcagacc
     1921 gtctacgccg acctcgttca acaggtccag ggcggcacgg atcactgtat tcggctgcaa
     1981 ctttgtcatg cttgacactt tatcactgat aaacataata tgtccaccaa cttatcagtg
     2041 ataaagaatc cgcgcgttca atcggaccag cggaggctgg tccggaggcc agacgtgaaa
     2101 cccaacatac ccctgatcgt aattctgagc actgtcgcgc tcgacgctgt cggcatcggc
     2161 ctgattatgc cggtgctgcc gggcctcctg cgcgatctgg ttcactcgaa cgacgtcacc
     2221 gcccactatg gcattctgct ggcgctgtat gcgttggtgc aatttgcctg cgcacctgtg
     2281 ctgggcgcgc tgtcggatcg tttcgggcgg cggccaatct tgctcgtctc gctggccggc
     2341 gccagatctg gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
     2401 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
     2461 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
     2521 tcatgagcgg agaattaagg gagtcacgtt atgacccccg ccgatgacgc gggacaagcc
     2581 gttttacgtt tggaactgac agaaccgcaa cgttgaagga gccactcagc cgcgggtttc
     2641 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
     2701 taaggtcact atcagctagc aaatatttct tgtcaaaaat gctccactga cgttccataa
     2761 attcccctcg gtatccaatt agagtctcat attcactctc aatccaaata atctgcaccg
     2821 gatctggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2881 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2941 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3001 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3061 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3121 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3181 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3241 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3301 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3361 aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3421 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3481 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3541 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3601 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3661 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3721 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3781 catgctggag ttcttcgccc acgggatctc tgcggaacag gcggtcgaag gtgccgatat
     3841 cattacgaca gcaacggccg acaagcacaa cgccacgatc ctgagcgaca atatgatcgg
     3901 gcccggcgtc cacatcaacg gcgtcggcgg cgactgccca ggcaagaccg agatgcaccg
     3961 cgatatcttg ctgcgttcgg atattttcgt ggagttcccg ccacagaccc ggatgatccc
     4021 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     4081 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     4141 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     4201 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     4261 tatgttacta gatcgggcct cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg
     4321 gcggctctga gggtggtggc tctgagggtg gcggttctga gggtggcggc tctgagggag
     4381 gcggttccgg tggtggctct ggttccggtg attttgatta tgaaaagatg gcaaacgcta
     4441 ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac gctaaaggca
     4501 aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt ggtgacgttt
     4561 ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat tcccaaatgg
     4621 ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa tatttacctt
     4681 ccctccctca atcggttgaa tgtcgccctt ttgtctttgg cccaatacgc aaaccgcctc
     4741 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4801 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4861 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4921 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc cccagattag
     4981 ccttttcaat ttcagaaaga atgctaaccc acagatggtt agagaggctt acgcagcagg
     5041 tctcatcaag acgatctacc cgagcaataa tctccaggaa atcaaatacc ttcccaagaa
     5101 ggttaaagat gcagtcaaaa gattcaggac taactgcatc aagaacacag agaaagatat
     5161 atttctcaag atcagaagta ctattccagt atggacgatt caaggcttgc ttcacaaacc
     5221 aaggcaagta atagagattg gagtctctaa aaaggtagtt cccactgaat caaaggccat
     5281 ggagtcaaag attcaaatag aggacctaac agaactcgcc gtaaagactg gcgaacagtt
     5341 catacagagt ctcttacgac tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca
     5401 cgacacactt gtctactcca aaaatatcaa agatacagtc tcagaagacc aaagggcaat
     5461 tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat
     5521 ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg
     5581 cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc
     5641 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
     5701 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
     5761 agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg ggactctaga
     5821 ggatccccgg gtggtcagtc ccttatgtta cgtcctgtag aaaccccaac ccgtgaaatc
     5881 aaaaaactcg acggcctgtg ggcattcagt ctggatcgcg aaaactgtgg aattgatcag
     5941 cgttggtggg aaagcgcgtt acaagaaagc cgggcaattg ctgtgccagg cagttttaac
     6001 gatcagttcg ccgatgcaga tattcgtaat tatgcgggca acgtctggta tcagcgcgaa
     6061 gtctttatac cgaaaggttg ggcaggccag cgtatcgtgc tgcgtttcga tgcggtcact
     6121 cattacggca aagtgtgggt caataatcag gaagtgatgg agcatcaggg cggctatacg
     6181 ccatttgaag ccgatgtcac gccgtatgtt attgccggga aaagtgtacg tatcaccgtt
     6241 tgtgtgaaca acgaactgaa ctggcagact atcccgccgg gaatggtgat taccgacgaa
     6301 aacggcaaga aaaagcagtc ttacttccat gatttcttta actatgccgg aatccatcgc
     6361 agcgtaatgc tctacaccac gccgaacacc tgggtggacg atatcaccgt ggtgacgcat
     6421 gtcgcgcaag actgtaacca cgcgtctgtt gactggcagg tggtggccaa tggtgatgtc
     6481 agcgttgaac tgcgtgatgc ggatcaacag gtggttgcaa ctggacaagg cactagcggg
     6541 actttgcaag tggtgaatcc gcacctctgg caaccgggtg aaggttatct ctatgaactg
     6601 tgcgtcacag ccaaaagcca gacagagtgt gatatctacc cgcttcgcgt cggcatccgg
     6661 tcagtggcag tgaagggcga acagttcctg attaaccaca aaccgttcta ctttactggc
     6721 tttggtcgtc atgaagatgc ggacttgcgt ggcaaaggat tcgataacgt gctgatggtg
     6781 cacgaccacg cattaatgga ctggattggg gccaactcct accgtacctc gcattaccct
     6841 tacgctgaag agatgctcga ctgggcagat gaacatggca tcgtggtgat tgatgaaact
     6901 gctgctgtcg gctttaacct ctctttaggc attggtttcg aagcgggcaa caagccgaaa
     6961 gaactgtaca gcgaagaggc agtcaacggg gaaactcagc aagcgcactt acaggcgatt
     7021 aaagagctga tagcgcgtga caaaaaccac ccaagcgtgg tgatgtggag tattgccaac
     7081 gaaccggata cccgtccgca aggtgcacgg gaatatttcg cgccactggc ggaagcaacg
     7141 cgtaaactcg acccgacgcg tccgatcacc tgcgtcaatg taatgttctg cgacgctcac
     7201 accgatacca tcagcgatct ctttgatgtg ctgtgcctga accgttatta cggatggtat
     7261 gtccaaagcg gcgatttgga aacggcagag aaggtactgg aaaaagaact tctggcctgg
     7321 caggagaaac tgcatcagcc gattatcatc accgaatacg gcgtggatac gttagccggg
     7381 ctgcactcaa tgtacaccga catgtggagt gaagagtatc agtgtgcatg gctggatatg
     7441 tatcaccgcg tctttgatcg cgtcagcgcc gtcgtcggtg aacaggtatg gaatttcgcc
     7501 gattttgcga cctcgcaagg catattgcgc gttggcggta acaagaaagg gatcttcact
     7561 cgcgaccgca aaccgaagtc ggcggctttt ctgctgcaaa aacgctggac tggcatgaac
     7621 ttcggtgaaa aaccgcagca gggaggcaaa caatgaatca acaactctcc tggcgcacca
     7681 tcgtcggcta cagcctcggg aattgctacc gagctcgaat ttccccgatc gttcaaacat
     7741 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
     7801 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat
     7861 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa
     7921 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg
     7981 ggaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     8041 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     8101 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcgc ccgctccttt cgctttcttc
     8161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     8281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8341 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     8401 tattcttttg atttataagg gattttgccg atttcggaac caccatcaaa caggattttc
     8461 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     8521 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccacccca gtacattaaa
     8581 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
     8641 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
     8701 acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag gcggcagact
     8761 ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg tttgaaacac
     8821 ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt gctgcctgtg
     8881 atcaaatatc atctccctcg cagagatccg aattatcagc cttcttattc atttctcgct
     8941 taaccgtgac aggctgtcga tcttgagaac tatgccgaca taataggaaa tcgctggata
     9001 aagccgctga ggaagctgag tggcgctatt tctttagaag tgaacgttga cgatatcaac
     9061 tcccctatcc attgctcacc gaatggtaca ggtcggggac ccgaagttcc gactgtcggc
     9121 ctgatgcatc cccggctgat cgaccccaga tctggggctg agaaagccca gtaaggaaac
     9181 aactgtaggt tcgagtcgcg agatcccccg gaaccaaagg aagtaggtta aacccgctcc
     9241 gatcaggccg agccacgcca ggccgagaac attggttcct gtaggcatcg ggattggcgg
     9301 atcaaacact aaagctactg gaacgagcag aagtcctccg gccgccagtt gccaggcggt
     9361 aaaggtgagc agaggcacgg gaggttgcca cttgcgggtc agcacggttc cgaacgccat
     9421 ggaaaccgcc cccgccaggc ccgctgcgac gccgacagga tctagcgctg cgtttggtgt
     9481 caacaccaac agcgccacgc ccgcagttcc gcaaatagcc cccaggaccg ccatcaatcg
     9541 tatcgggcta cctagcagag cggcagagat gaacacgacc atcagcggct gcacagcgcc
     9601 taccgtcgcc gcgaccccgc ccggcaggcg gtagaccgaa ataaacaaca agctccagaa
     9661 tagcgaaata ttaagtgcgc cgaggatgaa gatgcgcatc caccagattc ccgttggaat
     9721 ctgtcggacg atcatcacga gcaataaacc cgccggcaac gcccgcagca gcataccggc
     9781 gacccctcgg cctcgctgtt cgggctccac gaaaacgccg gacagatgcg ccttgtgagc
     9841 gtccttgggg ccgtcctcct gtttgaagac cgacagccca atgatctcgc cgtcgatgta
     9901 ggcgccgaat gccacggcat ctcgcaaccg ttcagcgaac gcctccatgg gctttttctc
     9961 ctcgtgctcg taaacggacc cgaacatctc tggagctttc ttcagggccg acaatcggat
    10021 ctcgcggaaa tcctgcacgt cggccgctcc aagccgtcga atctgagcct taatcacaat
    10081 tgtcaatttt aatcctctgt ttatcggcag ttcgtagagc gcgccgtgcg tcccgagcga
    10141 tactgagcga agcaagtgcg tcgagcagtg cccgcttgtt cctgaaatgc cagtaaagcg
    10201 ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc
    10261 aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct
    10321 tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg
    10381 aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt
    10441 cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca
    10501 gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca
    10561 cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg
    10621 gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa
    10681 taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc
    10741 ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca
    10801 tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa
    10861 atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca
    10921 gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac
    10981 aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg
    11041 gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata
    11101 gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcgagacga
    11161 cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg
    11221 tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg gaaggtttcg
    11281 cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc
    11341 gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct
    11401 tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt tgacccgcct
    11461 ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc gtagaccgtc
    11521 tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac cagcgacccc
    11581 ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat gcggaagaag
    11641 tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggtact aaaacaattc
    11701 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
    11761 aaatagctcg acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc
    11821 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
    11881 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
    11941 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
    12001 ttcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
    12061 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
    12121 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
    12181 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
    12241 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
    12301 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
    12361 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
    12421 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
    12481 catttattat ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat
    12541 aacaagacga actccaattc actgttcctt gcattctaaa accttaaata ccagaaaaca
    12601 gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga gccgattttg
    12661 aaaccacaat tatgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga
    12721 ggtgctccag tggcttctgt gtctatcagc tgtccctcct gttcagctac tgacggggtg
    12781 gtgcgtaacg gcaaaagcac cgccggacat cagcgctatc tctgctctca ctgccgtaaa
    12841 acatggcaac tgcagttcac ttacaccgct tctcaacccg gtacgcacca gaaaatcatt
    12901 gatatggcca tgaatggcgt tggatgccgg gcaacagccc gcattatggg cgttggcctc
    12961 aacacgattt tacgtcactt aaaaaactca ggccgcagtc ggtaacctcg cgcatacagc
    13021 cgggcagtga cgtcatcgtc tgcgcggaaa tggacgaaca gtggggctat gtcggggcta
    13081 aatcgcgcca gcgctggctg ttttacgcgt atgacagtct ccggaagacg gttgttgcgc
    13141 acgtattcgg tgaacgcact atggcgacgc tggggcgtct tatgagcctg ctgtcaccct
    13201 ttgacgtggt gatatggatg acggatggct ggccgctgta tgaatcccgc ctgaagggaa
    13261 agctgcacgt aatcagcaag cgatatacgc agcgaattga gcggcataac ctgaatctga
    13321 ggcagcacct ggcacggctg ggacggaagt cgctgtcgtt ctcaaaatcg gtggagctgc
    13381 atgacaaagt catcgggcat tatctgaaca taaaacacta tcaataagtt ggagtcatta
    13441 cccaattatg atagaattta caagctataa ggttattgtc ctgggtttca agcattagtc
    13501 catgcaagtt tttatgcttt gcccattcta tagatatatt gataagcgcg ctgcctatgc
    13561 cttgccccct gaaatcctta catacggcga tatcttctat ataaaagata tattatctta
    13621 tcagtattgt caatatattc aaggcaatct gcctcctcat cctcttcatc ctcttcgtct
    13681 tggtagcttt ttaaatatgg cgcttcatag agtaattctg taaaggtcca attctcgttt
    13741 tcatacctcg gtataatctt acctatcacc tcaaatggtt cgctgggttt atcgcacccc
    13801 cgaacacgag cacggcaccc gcgaccacta tgccaagaat gcccaaggta aaaattgccg
    13861 gccccgccat gaagtccgtg aatgccccga cggccgaagt gaagggcagg ccgccaccca
    13921 ggccgccgcc ctcactgccc ggcacctggt cgctgaatgt cgatgccagc acctgcggca
    13981 cgtcaatgct tccgggcgtc gcgctcgggc tgatcgccca tcccgttact gccccgatcc
    14041 cggcaatggc aaggactgcc agcgctgcca tttttggggt gaggccgttc gcggccgagg
    14101 ggcgcagccc ctggggggat gggaggcccg cgttagcggg ccgggagggt tcgagaaggg
    14161 ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg cgcagccctg gttaaaaaca
    14221 aggtttataa atattggttt aaaagcaggt taaaagacag gttagcggtg gccgaaaaac
    14281 gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg acagcccctc aaatgtcaat
    14341 aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg tcaaggatcg cgcccctcat
    14401 ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg cacttatccc caggcttgtc
    14461 cacatcatct gtgggaaact cgcgtaaaat caggcgtttt cgccgatttg cgaggctggc
    14521 cagctccacg tcgccggccg aaatcgagcc tgcccctcat ctgtcaacgc cgcgccgggt
    14581 gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt cagtgagggc caagttttcc
    14641 gcgaggtatc cacaacgccg gcggccgcgg tgtctcgcac acggcttcga cggcgtttct
    14701 ggcgcgtttg cagggccata gacggccgcc agcccagcgg cgagggcaac cagcccgg