LOCUS KL-pGEn3-DEST 8949 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers CDS 129..1108 /dnas_title="AmpR" /label=AmpR /vntifkey="4" rep_origin 1137..1804 /dnas_title="Col E1 origin" /label=Col E1 origin /vntifkey="33" misc_feature 1998..2127 /note="5'ITR" misc_feature 2208..2790 /note="CMV Promoter" misc_feature 2828..2960 /note="Artificial Intron" misc_recomb complement(4541..4569) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 7577..8118 /note="WPRE" misc_recomb 7539..7561 /label=attB2 /note="attB2 site" /note="recognized as attB2" misc_feature complement(8125..8339) /note="BGH pA" misc_feature complement(8427..8556) /note="3'-ITR" misc_feature 2982..3056 /note="TCM signal sequence" misc_feature 3060..3083 /note="His tag" misc_feature 3084..3134 /note="AviTag" misc_feature 3135..3830 /note="Superfolder GFP" misc_feature 3837..4532 /note="Fc domain" misc_feature 2976..2981 /note="Kozak seq" misc_feature 4570..4592 /note="TEV site" source 4593..7532 /dnas_title="KL-pGEn3-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /tissue_type="kidney" gene 4593..7532 /dnas_title="klotho" /gene="klotho" CDS 4593..7532 /dnas_title="klotho" /gene="klotho" /codon_start=1 /protein_id="BAA23382.1" /db_xref="GI:2618596" /translation="MPASAPPRRPRPPPPSLSLLLVLLGLGGRRLRAEPGDGAQTWAR VSRPPAPEAAGLFQGTFPDGFLWAVGSAAYQTEGGWQQHGKGASIWDTFTHHPLAPPG DSRNASLPLGAPSPLQPATGDVASDSYNNVFRDTEALRELGVTHYRFSISWARVLPNG SAGVPNREGLRYYRRLLERLRELGVQPVVTLYHWDLPQRLQDAYGGWANRALADHFRD YAELCFRHFGGQVKYWITIDNPYVVAWHGYATGRLAPGIRGSPRLGYLVAHNLLLAHA KVWHLYNTSFRPTQGGQVSIALSSHWINPRRMTDHSIKECQKSLDFVLGWFAKPVFID GDYPESMKNNLSSILPDFTESEKKFIKGTADFFALCFGPTLSFQLLDPHMKFRQLESP NLRQLLSWIDLEFNHPQIFIVENGWFVSGTTKRDDAKYMYYLKKFIMETLKAIKLDGV DVIGYTAWSLMDGFEWHRGYSIRRGLFYVDFLSQDKMLLPKSSALFYQKLIEKNGFPP LPENQPLEGTFPCDFAWGVVDNYIQVDTTLSQFTDLNVYLWDVHHSKRLIKVDGVVTK KRKSYCVDFAAIQPQIALLQEMHVTHFRFSLDWALILPLGNQSQVNHTILQYYRCMAS ELVRVNITPVVALWQPMAPNQGLPRLLARQGAWENPYTALAFAEYARLCFQELGHHVK LWITMNEPYTRNMTYSAGHNLLKAHALAWHVYNEKFRHAQNGKISIALQADWIEPACP FSQKDKEVAERVLEFDIGWLAEPIFGSGDYPWVMRDWLNQRNNFLLPYFTEDEKKLIQ GTFDFLALSHYTTILVDSEKEDPIKYNDYLEVQEMTDITWLNSPSQVAVVPWGLRKVL NWLKFKYGDLPMYIISNGIDDGLHAEDDQLRVYYMQNYINEALKAHILDGINLCGYFA YSFNDRTAPRFGLYRYAADQFEPKASMKHYRKIIDSNGFPGPETLERFCPEEFTVCTE CSFFHTRKSLLAFIAFLFFASIISLSLIFYYSKKGRRSYK" misc_feature 4593..7532 /note="KL coding region" vector join(7547..8949,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..7546 /source="KL-Nterm-TEVfusion-pDONR221" /type="Entry Clone" ORIGIN 1 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT 61 CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA 121 GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT 181 GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT 241 TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT 301 TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG 361 TATTATCCCG TATTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA 421 ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA 481 GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA 541 CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA 601 CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA 661 CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA 721 CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC 781 TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC 841 GTGGTTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG 901 TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA 961 TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT 1021 AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA 1081 ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG 1141 AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA 1201 CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT 1261 TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTTCTT CTAGTGTAGC 1321 CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA 1381 TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA 1441 GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC 1501 CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CTATGAGAAA 1561 GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA 1621 CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG 1681 GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC 1741 TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG 1801 CTCATTAGGC ACCCCAGGCT TTACCCGAAC GACCGAGCGC AGCGAGTCAG TGAGCGAGGA 1861 AGCGGAGAGC GCCCAATACG CAAGGAAACA GCTATGACCA TGTTAATGCA GCTGGCACGA 1921 CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGGAAGGCC CATGAGGCCA GTTAATTAAG 1981 AGGTACCTTA ATTAAGGCTG CGCGCTCGCT CGCTCACTGA GGCCGCCCGG GCAAAGCCCG 2041 GGCGTCGGGC GACCTTTGGT CGCCCGGCCT CAGTGAGCGA GCGAGCGCGC AGAGAGGGAG 2101 TGGCCAACTC CATCACTAGG GGTTCCTTGT AGTTAATGAT TAACCCGCCA TGCTACTTAT 2161 CTACGTAGCC ATGCTCTAGG AAGATCGCAA TTCGCCCTTA AGCTAGCTAG TTATTAATAG 2221 TAATCAATTA CGGGGTCATT AGTTCATAGC CCATATATGG AGTTCCGCGT TACATAACTT 2281 ACGGTAAATG GCCCGCCTGG CTGACCGCCC AACGACCCCC GCCCATTGAC GTCAATAATG 2341 ACGTATGTTC CCATAGTAAC GCCAATAGGG ACTTTCCATT GACGTCAATG GGTGGAGTAT 2401 TTACGGTAAA CTGCCCACTT GGCAGTACAT CAAGTGTATC ATATGCCAAG TACGCCCCCT 2461 ATTGACGTCA ATGACGGTAA ATGGCCCGCC TGGCATTATG CCCAGTACAT GACCTTATGG 2521 GACTTTCCTA CTTGGCAGTA CATCTACGTA TTAGTCATCG CTATTACCAT GGTGATGCGG 2581 TTTTGGCAGT ACATCAATGG GCGTGGATAG CGGTTTGACT CACGGGGATT TCCAAGTCTC 2641 CACCCCATTG ACGTCAATGG GAGTTTGTTT TGGCACCAAA ATCAACGGGA CTTTCCAAAA 2701 TGTCGTAACA ACTCCGCCCC ATTGACGCAA ATGGGCGGTA GGCGTGTACG GTGGGAGGTC 2761 TATATAAGCA GAGCTGGTTT AGTGAACCGT CAGATCCTGC AGAAGTTGGT CGTGAGGCAC 2821 TGGGCAGGTA AGTATCAAGG TTACAAGACA GGTTTAAGGA GACCAATAGA AACTGGGCTT 2881 GTCGAGACAG AGAAGACTCT TGCGTTTCTG ATAGGCACCT ATTGGTCTTA CTGACATCCA 2941 CTTTGCCTTT CTCTCCACAG GTGTCCAgcg gccgcGCCAC cATGAGACTG CTGACCGCCC 3001 TGTTCGCCTA CTTCATCGTG GCCCTGATCC TGGCCTTCAG CGTGTCCGCC AAGAGCATGC 3061 ACCACCACCA TCACCATCAT CACATGAGCG GCCTGAACGA CATCTTCGAG GCCCAGAAAA 3121 TCGAGTGGCA CGAGATGAGC AAGGGCGAGG AACTGTTCAC CGGCGTGGTG CCCATCCTGG 3181 TGGAACTGGA CGGCGACGTG AACGGCCACA AGTTCTCTGT GCGGGGCGAG GGCGAGGGGG 3241 ACGCCACAAA TGGCAAGCTG ACCCTGAAGT TCATCTGCAC CACCGGAAAG CTGCCCGTGC 3301 CCTGGCCTAC CCTGGTCACA ACCCTGACCT ACGGCGTGCA GTGCTTCAGC AGATACCCCG 3361 ACCACATGAA GCGGCACGAT TTCTTCAAGA GCGCCATGCC CGAGGGCTAC GTGCAAGAAC 3421 GGACCATCAG CTTCAAGGAC GACGGCACCT ACAAGACCAG AGCCGAAGTG AAGTTCGAGG 3481 GCGACACCCT GGTCAACCGG ATCGAGCTGA AGGGCATCGA CTTCAAAGAG GACGGCAACA 3541 TCCTGGGCCA CAAGCTCGAG TACAACTTCA ACAGCCACAA CGTGTACATC ACCGCCGACA 3601 AGCAGAAGAA CGGCATCAAG GCCAACTTCA AGATCCGGCA CAACGTGGAA GATGGCAGCG 3661 TGCAGCTGGC CGACCACTAC CAGCAGAACA CCCCCATCGG CGACGGCCCC GTGCTGCTGC 3721 CCGACAATCA CTACCTGAGC ACCCAGAGCG TGCTGAGCAA GGACCCCAAC GAGAAGCGGG 3781 ACCACATGGT GCTGCTGGAA TTTGTGACCG CCGCTGGCAT CACCCACGGC GAATTCgagc 3841 ccaaatcttg tgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg 3901 gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc 3961 ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact 4021 ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca 4081 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca 4141 aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct 4201 ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggatg 4261 agctgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca 4321 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg 4381 tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt 4441 ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga ggctctgcac aaccactaca 4501 cgcagaagag cctctccctg tctccgggta aagaattcgc ATCAACAAGT TTGTACAAAA 4561 AAGCAGGCTc tgaaaacttg tactttcaag gcgcggagcc gggcgacggc gcgcagacct 4621 gggcccgtgt ctcgcggcct cctgcccccg aggccgcggg cctcttccag ggcaccttcc 4681 ccgacggctt cctctgggcc gtgggcagcg ccgcctacca gaccgagggc ggctggcagc 4741 agcacggcaa gggtgcgtcc atctgggaca cgttcaccca ccaccccctg gcacccccgg 4801 gagactcccg gaacgccagt ctgccgttgg gcgccccgtc gccgctgcag cccgccaccg 4861 gggacgtagc cagcgacagc tacaacaacg tcttccgcga cacggaggcg ctgcgcgagc 4921 tcggggtcac tcactaccgc ttctccatct cgtgggcgcg agtgctcccc aatggcagcg 4981 cgggcgtccc caaccgcgag gggctgcgct actaccggcg cctgctggag cggctgcggg 5041 agctgggcgt gcagcccgtg gtcaccctgt accactggga cctgccccag cgcctgcagg 5101 acgcctacgg cggctgggcc aaccgcgccc tggccgacca cttcagggat tacgcggagc 5161 tctgcttccg ccacttcggc ggtcaggtca agtactggat caccatcgac aacccctacg 5221 tggtggcctg gcacggctac gccaccgggc gcctggcccc cggcatccgg ggcagcccgc 5281 ggctcgggta cctggtggcg cacaacctcc tcctggctca tgccaaagtc tggcatctct 5341 acaatacttc tttccgtccc actcagggag gtcaggtgtc cattgcccta agctctcact 5401 ggatcaatcc tcgaagaatg accgaccaca gcatcaaaga atgtcaaaaa tctctggact 5461 ttgtactagg ttggtttgcc aaacccgtat ttattgatgg tgactatccc gagagcatga 5521 agaataacct ttcatctatt ctgcctgatt ttactgaatc tgagaaaaag ttcatcaaag 5581 gaactgctga cttttttgct ctttgctttg gacccacctt gagttttcaa cttttggacc 5641 ctcacatgaa gttccgccaa ttggaatctc ccaacctgag gcaactgctt tcctggattg 5701 accttgaatt taaccatcct caaatattta ttgtggaaaa tggctggttt gtctcaggga 5761 ccaccaagag agatgatgcc aaatatatgt attacctcaa aaagttcatc atggaaacct 5821 taaaagccat caagctggat ggggtggatg tcatcgggta taccgcatgg tccctcatgg 5881 atggtttcga gtggcacaga ggttacagca tcaggcgtgg actcttctat gttgactttc 5941 taagccagga caagatgttg ttgccaaagt cttcagcctt gttctaccaa aagctgatag 6001 agaaaaatgg cttccctcct ttacctgaaa atcagcccct agaagggaca tttccctgtg 6061 actttgcttg gggagttgtt gacaactaca ttcaagtaga taccactctg tctcagttta 6121 ccgacctgaa tgtttacctg tgggatgtcc accacagtaa aaggcttatt aaagtggatg 6181 gggttgtgac caagaagagg aaatcctact gtgttgactt tgctgccatc cagccccaga 6241 tcgctttact ccaggaaatg cacgttacac attttcgctt ctccctggac tgggccctga 6301 ttctccctct gggtaaccag tcccaggtga accacaccat cctgcagtac tatcgctgca 6361 tggccagcga gcttgtccgt gtcaacatca ccccagtggt ggccctgtgg cagcctatgg 6421 ccccgaacca aggactgccg cgcctcctgg ccaggcaggg cgcctgggag aacccctaca 6481 ctgccctggc ctttgcagag tatgcccgac tgtgctttca agagctcggc catcacgtca 6541 agctttggat aacgatgaat gagccgtata caaggaatat gacatacagt gctggccaca 6601 accttctgaa ggcccatgcc ctggcttggc atgtgtacaa tgaaaagttt aggcatgctc 6661 agaatgggaa aatatccata gccttgcagg ctgattggat agaacctgcc tgccctttct 6721 cccaaaagga caaagaggtg gccgagagag ttttggaatt tgacattggc tggctggctg 6781 agcccatttt cggctctgga gattatccat gggtgatgag ggactggctg aaccaaagaa 6841 acaattttct tcttccttat ttcactgaag atgaaaaaaa gctaatccag ggtacctttg 6901 actttttggc tttaagccat tataccacca tccttgtaga ctcagaaaaa gaagatccaa 6961 taaaatacaa tgattaccta gaagtgcaag aaatgaccga catcacgtgg ctcaactccc 7021 ccagtcaggt ggcggtagtg ccctgggggt tgcgcaaagt gctgaactgg ctgaagttca 7081 agtacggaga cctccccatg tacataatat ccaacggaat cgatgacggg ctgcatgctg 7141 aggacgacca gctgagggtg tattatatgc agaattacat aaacgaagct ctcaaagccc 7201 acatactgga tggtatcaat ctttgcggat actttgctta ttcgtttaac gaccgcacag 7261 ctccgaggtt tggcctctat cgttatgctg cagatcagtt tgagcccaag gcatccatga 7321 aacattacag gaaaattatt gacagcaatg gtttcccggg cccagaaact ctggaaagat 7381 tttgtccaga agaattcacc gtgtgtactg agtgcagttt ttttcacacc cgaaagtctt 7441 tactggcttt catagctttt ctattttttg cttctattat ttctctctcc cttatatttt 7501 actactcgaa gaaaggcaga agaagttaca aaTAGGACCC AGCTTTCTTG TACAAAGTGG 7561 TTGATAGCTT GGATCCAATC AACCTCTGGA TTACAAAATT TGTGAAAGAT TGACTGGTAT 7621 TCTTAACTAT GTTGCTCCTT TTACGCTATG TGGATACGCT GCTTTAATGC CTTTGTATCA 7681 TGCTATTGCT TCCCGTATGG CTTTCATTTT CTCCTCCTTG TATAAATCCT GGTTGCTGTC 7741 TCTTTATGAG GAGTTGTGGC CCGTTGTCAG GCAACGTGGC GTGGTGTGCA CTGTGTTTGC 7801 TGACGCAACC CCCACTGGTT GGGGCATTGC CACCACCTGT CAGCTCCTTT CCGGGACTTT 7861 CGCTTTCCCC CTCCCTATTG CCACGGCGGA ACTCATCGCC GCCTGCCTTG CCCGCTGCTG 7921 GACAGGGGCT CGGCTGTTGG GCACTGACAA TTCCGTGGTG TTGTCGGGGA AGCTGACGTC 7981 CTTTCCATGG CTGCTCGCCT GTGTTGCCAC CTGGATTCTG CGCGGGACGT CCTTCTGCTA 8041 CGTCCCTTCG GCCCTCAATC CAGCGGACCT TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG 8101 GCCTCTTCCG CGTCTTCGAG ATCTGCCTCG ACTGTGCCTT CTAGTTGCCA GCCATCTGTT 8161 GTTTGCCCCT CCCCCGTGCC TTCCTTGACC CTGGAAGGTG CCACTCCCAC TGTCCTTTCC 8221 TAATAAAATG AGGAAATTGC ATCGCATTGT CTGAGTAGGT GTCATTCTAT TCTGGGGGGT 8281 GGGGTGGGGC AGGACAGCAA GGGGGAGGAT TGGGAAGACA ATAGCAGGCA TGCTGGGGAC 8341 TCGAGTTAAG GGCGAATTGC CGATTAGGAT CTTCCTAGAG CATGGCTACG TAGATAAGTA 8401 GCATGGCGGG TTAATCATTA ACTACAAGGA ACCCCTAGTG ATGGAGTTGG CCACTCCCTC 8461 TCTGCGCGCT CGCTCGCTCA CTGAGGCCGG GCGACCAAAG GTCGCCCGAC GCCCGGGCTT 8521 TGCCCGGGCG GCCTCAGTGA GCGAGCGAGC GCGCAGCCTT AATTAAGAGC TCATGGCGCG 8581 CCTAGGCCTT GACGGCCTTC CGCCAATTCG CCCTATAGTG AGTCGTATTA CGTCGCGCTC 8641 ACTGGCCGTC GTTTTACAAC GTCGTGACTG GGAAAACCCT GGCGTTACCC AACTTAATCG 8701 CCTTGCAGCA CATCCCCCTT TCGCCAGCTG GCGTAATAGC GAAGAGGCCC GCACCGAAAC 8761 GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGGA GCGCCCTGTA GCGGCCACTC 8821 AACCCTATCT CGGTCTATTC TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG 8881 TTAAAAAATG AGCTGATTTA ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGCTT 8941 ACAATTTAG //