LOCUS KL-pGEn1-DEST 7522 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_feature 2977..3054 /note="TCM Sig Seq" misc_recomb complement(3114..3142) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 3055..3078 /note="8xHis Tag" misc_feature 3085..3108 /note="StrepTag" misc_feature 6150..6691 /note="WPRE" misc_recomb 6112..6134 /label=attB2 /note="attB2 site" /note="recognized as attB2" misc_feature complement(6698..6912) /note="BGH pA" misc_feature complement(7000..7129) /note="3'-ITR" misc_feature 3143..3165 /note="TEV site" source 3166..6105 /dnas_title="KL-pGEn1-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /tissue_type="kidney" gene 3166..6105 /dnas_title="klotho" /gene="klotho" CDS 3166..6105 /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 3166..6105 /note="KL coding region" vector join(6120..7522,1..3132) /source="pGEn1-DEST" /type="Destination Vector" clone 3133..6119 /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 GCCGCCATGA GACTGCTGAC AGCCCTGTTC 3001 GCCTACTTCA TCGTGGCCCT GATCCTGGCC TTCAGCGTGT CCGCCAAGAG CATGCACCAC 3061 CACCATCACC ATCATCACAG CGCCTGGTCC CACCCCCAGT TCGAGAAAGA ATTATCAACA 3121 AGTTTGTACA AAAAAGCAGG CTctgaaaac ttgtactttc aaggcgcgga gccgggcgac 3181 ggcgcgcaga cctgggcccg tgtctcgcgg cctcctgccc ccgaggccgc gggcctcttc 3241 cagggcacct tccccgacgg cttcctctgg gccgtgggca gcgccgccta ccagaccgag 3301 ggcggctggc agcagcacgg caagggtgcg tccatctggg acacgttcac ccaccacccc 3361 ctggcacccc cgggagactc ccggaacgcc agtctgccgt tgggcgcccc gtcgccgctg 3421 cagcccgcca ccggggacgt agccagcgac agctacaaca acgtcttccg cgacacggag 3481 gcgctgcgcg agctcggggt cactcactac cgcttctcca tctcgtgggc gcgagtgctc 3541 cccaatggca gcgcgggcgt ccccaaccgc gaggggctgc gctactaccg gcgcctgctg 3601 gagcggctgc gggagctggg cgtgcagccc gtggtcaccc tgtaccactg ggacctgccc 3661 cagcgcctgc aggacgccta cggcggctgg gccaaccgcg ccctggccga ccacttcagg 3721 gattacgcgg agctctgctt ccgccacttc ggcggtcagg tcaagtactg gatcaccatc 3781 gacaacccct acgtggtggc ctggcacggc tacgccaccg ggcgcctggc ccccggcatc 3841 cggggcagcc cgcggctcgg gtacctggtg gcgcacaacc tcctcctggc tcatgccaaa 3901 gtctggcatc tctacaatac ttctttccgt cccactcagg gaggtcaggt gtccattgcc 3961 ctaagctctc actggatcaa tcctcgaaga atgaccgacc acagcatcaa agaatgtcaa 4021 aaatctctgg actttgtact aggttggttt gccaaacccg tatttattga tggtgactat 4081 cccgagagca tgaagaataa cctttcatct attctgcctg attttactga atctgagaaa 4141 aagttcatca aaggaactgc tgactttttt gctctttgct ttggacccac cttgagtttt 4201 caacttttgg accctcacat gaagttccgc caattggaat ctcccaacct gaggcaactg 4261 ctttcctgga ttgaccttga atttaaccat cctcaaatat ttattgtgga aaatggctgg 4321 tttgtctcag ggaccaccaa gagagatgat gccaaatata tgtattacct caaaaagttc 4381 atcatggaaa ccttaaaagc catcaagctg gatggggtgg atgtcatcgg gtataccgca 4441 tggtccctca tggatggttt cgagtggcac agaggttaca gcatcaggcg tggactcttc 4501 tatgttgact ttctaagcca ggacaagatg ttgttgccaa agtcttcagc cttgttctac 4561 caaaagctga tagagaaaaa tggcttccct cctttacctg aaaatcagcc cctagaaggg 4621 acatttccct gtgactttgc ttggggagtt gttgacaact acattcaagt agataccact 4681 ctgtctcagt ttaccgacct gaatgtttac ctgtgggatg tccaccacag taaaaggctt 4741 attaaagtgg atggggttgt gaccaagaag aggaaatcct actgtgttga ctttgctgcc 4801 atccagcccc agatcgcttt actccaggaa atgcacgtta cacattttcg cttctccctg 4861 gactgggccc tgattctccc tctgggtaac cagtcccagg tgaaccacac catcctgcag 4921 tactatcgct gcatggccag cgagcttgtc cgtgtcaaca tcaccccagt ggtggccctg 4981 tggcagccta tggccccgaa ccaaggactg ccgcgcctcc tggccaggca gggcgcctgg 5041 gagaacccct acactgccct ggcctttgca gagtatgccc gactgtgctt tcaagagctc 5101 ggccatcacg tcaagctttg gataacgatg aatgagccgt atacaaggaa tatgacatac 5161 agtgctggcc acaaccttct gaaggcccat gccctggctt ggcatgtgta caatgaaaag 5221 tttaggcatg ctcagaatgg gaaaatatcc atagccttgc aggctgattg gatagaacct 5281 gcctgccctt tctcccaaaa ggacaaagag gtggccgaga gagttttgga atttgacatt 5341 ggctggctgg ctgagcccat tttcggctct ggagattatc catgggtgat gagggactgg 5401 ctgaaccaaa gaaacaattt tcttcttcct tatttcactg aagatgaaaa aaagctaatc 5461 cagggtacct ttgacttttt ggctttaagc cattatacca ccatccttgt agactcagaa 5521 aaagaagatc caataaaata caatgattac ctagaagtgc aagaaatgac cgacatcacg 5581 tggctcaact cccccagtca ggtggcggta gtgccctggg ggttgcgcaa agtgctgaac 5641 tggctgaagt tcaagtacgg agacctcccc atgtacataa tatccaacgg aatcgatgac 5701 gggctgcatg ctgaggacga ccagctgagg gtgtattata tgcagaatta cataaacgaa 5761 gctctcaaag cccacatact ggatggtatc aatctttgcg gatactttgc ttattcgttt 5821 aacgaccgca cagctccgag gtttggcctc tatcgttatg ctgcagatca gtttgagccc 5881 aaggcatcca tgaaacatta caggaaaatt attgacagca atggtttccc gggcccagaa 5941 actctggaaa gattttgtcc agaagaattc accgtgtgta ctgagtgcag tttttttcac 6001 acccgaaagt ctttactggc tttcatagct tttctatttt ttgcttctat tatttctctc 6061 tcccttatat tttactactc gaagaaaggc agaagaagtt acaaaTAGGA CCCAGCTTTC 6121 TTGTACAAAG TGGTTGATAG CTTGGATCCA ATCAACCTCT GGATTACAAA ATTTGTGAAA 6181 GATTGACTGG TATTCTTAAC TATGTTGCTC CTTTTACGCT ATGTGGATAC GCTGCTTTAA 6241 TGCCTTTGTA TCATGCTATT GCTTCCCGTA TGGCTTTCAT TTTCTCCTCC TTGTATAAAT 6301 CCTGGTTGCT GTCTCTTTAT GAGGAGTTGT GGCCCGTTGT CAGGCAACGT GGCGTGGTGT 6361 GCACTGTGTT TGCTGACGCA ACCCCCACTG GTTGGGGCAT TGCCACCACC TGTCAGCTCC 6421 TTTCCGGGAC TTTCGCTTTC CCCCTCCCTA TTGCCACGGC GGAACTCATC GCCGCCTGCC 6481 TTGCCCGCTG CTGGACAGGG GCTCGGCTGT TGGGCACTGA CAATTCCGTG GTGTTGTCGG 6541 GGAAGCTGAC GTCCTTTCCA TGGCTGCTCG CCTGTGTTGC CACCTGGATT CTGCGCGGGA 6601 CGTCCTTCTG CTACGTCCCT TCGGCCCTCA ATCCAGCGGA CCTTCCTTCC CGCGGCCTGC 6661 TGCCGGCTCT GCGGCCTCTT CCGCGTCTTC GAGATCTGCC TCGACTGTGC CTTCTAGTTG 6721 CCAGCCATCT GTTGTTTGCC CCTCCCCCGT GCCTTCCTTG ACCCTGGAAG GTGCCACTCC 6781 CACTGTCCTT TCCTAATAAA ATGAGGAAAT TGCATCGCAT TGTCTGAGTA GGTGTCATTC 6841 TATTCTGGGG GGTGGGGTGG GGCAGGACAG CAAGGGGGAG GATTGGGAAG ACAATAGCAG 6901 GCATGCTGGG GACTCGAGTT AAGGGCGAAT TGCCGATTAG GATCTTCCTA GAGCATGGCT 6961 ACGTAGATAA GTAGCATGGC GGGTTAATCA TTAACTACAA GGAACCCCTA GTGATGGAGT 7021 TGGCCACTCC CTCTCTGCGC GCTCGCTCGC TCACTGAGGC CGGGCGACCA AAGGTCGCCC 7081 GACGCCCGGG CTTTGCCCGG GCGGCCTCAG TGAGCGAGCG AGCGCGCAGC CTTAATTAAG 7141 AGCTCATGGC GCGCCTAGGC CTTGACGGCC TTCCGCCAAT TCGCCCTATA GTGAGTCGTA 7201 TTACGTCGCG CTCACTGGCC GTCGTTTTAC AACGTCGTGA CTGGGAAAAC CCTGGCGTTA 7261 CCCAACTTAA TCGCCTTGCA GCACATCCCC CTTTCGCCAG CTGGCGTAAT AGCGAAGAGG 7321 CCCGCACCGA AACGCCCTTC CCAACAGTTG CGCAGCCTGA ATGGCGAATG GGAGCGCCCT 7381 GTAGCGGCCA CTCAACCCTA TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT 7441 TTCGGCCTAT TGGTTAAAAA ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA 7501 AATATTAACG CTTACAATTT AG //