LOCUS KL-pGEn2-DEST 8247 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(3839..3867) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 6875..7416 /note="WPRE" misc_recomb 6837..6859 /label=attB2 /note="attB2 site" /note="recognized as attB2" misc_feature complement(7423..7637) /note="BGH pA" misc_feature complement(7725..7854) /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 2976..2981 /note="Kozak seq" misc_feature 3868..3890 /note="TEV site" source 3891..6830 /dnas_title="KL-pGEn2-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /tissue_type="kidney" gene 3891..6830 /dnas_title="klotho" /gene="klotho" CDS 3891..6830 /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 3891..6830 /note="KL coding region" vector join(6845..8247,1..3857) /source="pGEn2-DEST" /type="Destination Vector" clone 3858..6844 /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 GAATTCgcAT 3841 CAACAAGTTT GTACAAAAAA GCAGGCTctg aaaacttgta ctttcaaggc gcggagccgg 3901 gcgacggcgc gcagacctgg gcccgtgtct cgcggcctcc tgcccccgag gccgcgggcc 3961 tcttccaggg caccttcccc gacggcttcc tctgggccgt gggcagcgcc gcctaccaga 4021 ccgagggcgg ctggcagcag cacggcaagg gtgcgtccat ctgggacacg ttcacccacc 4081 accccctggc acccccggga gactcccgga acgccagtct gccgttgggc gccccgtcgc 4141 cgctgcagcc cgccaccggg gacgtagcca gcgacagcta caacaacgtc ttccgcgaca 4201 cggaggcgct gcgcgagctc ggggtcactc actaccgctt ctccatctcg tgggcgcgag 4261 tgctccccaa tggcagcgcg ggcgtcccca accgcgaggg gctgcgctac taccggcgcc 4321 tgctggagcg gctgcgggag ctgggcgtgc agcccgtggt caccctgtac cactgggacc 4381 tgccccagcg cctgcaggac gcctacggcg gctgggccaa ccgcgccctg gccgaccact 4441 tcagggatta cgcggagctc tgcttccgcc acttcggcgg tcaggtcaag tactggatca 4501 ccatcgacaa cccctacgtg gtggcctggc acggctacgc caccgggcgc ctggcccccg 4561 gcatccgggg cagcccgcgg ctcgggtacc tggtggcgca caacctcctc ctggctcatg 4621 ccaaagtctg gcatctctac aatacttctt tccgtcccac tcagggaggt caggtgtcca 4681 ttgccctaag ctctcactgg atcaatcctc gaagaatgac cgaccacagc atcaaagaat 4741 gtcaaaaatc tctggacttt gtactaggtt ggtttgccaa acccgtattt attgatggtg 4801 actatcccga gagcatgaag aataaccttt catctattct gcctgatttt actgaatctg 4861 agaaaaagtt catcaaagga actgctgact tttttgctct ttgctttgga cccaccttga 4921 gttttcaact tttggaccct cacatgaagt tccgccaatt ggaatctccc aacctgaggc 4981 aactgctttc ctggattgac cttgaattta accatcctca aatatttatt gtggaaaatg 5041 gctggtttgt ctcagggacc accaagagag atgatgccaa atatatgtat tacctcaaaa 5101 agttcatcat ggaaacctta aaagccatca agctggatgg ggtggatgtc atcgggtata 5161 ccgcatggtc cctcatggat ggtttcgagt ggcacagagg ttacagcatc aggcgtggac 5221 tcttctatgt tgactttcta agccaggaca agatgttgtt gccaaagtct tcagccttgt 5281 tctaccaaaa gctgatagag aaaaatggct tccctccttt acctgaaaat cagcccctag 5341 aagggacatt tccctgtgac tttgcttggg gagttgttga caactacatt caagtagata 5401 ccactctgtc tcagtttacc gacctgaatg tttacctgtg ggatgtccac cacagtaaaa 5461 ggcttattaa agtggatggg gttgtgacca agaagaggaa atcctactgt gttgactttg 5521 ctgccatcca gccccagatc gctttactcc aggaaatgca cgttacacat tttcgcttct 5581 ccctggactg ggccctgatt ctccctctgg gtaaccagtc ccaggtgaac cacaccatcc 5641 tgcagtacta tcgctgcatg gccagcgagc ttgtccgtgt caacatcacc ccagtggtgg 5701 ccctgtggca gcctatggcc ccgaaccaag gactgccgcg cctcctggcc aggcagggcg 5761 cctgggagaa cccctacact gccctggcct ttgcagagta tgcccgactg tgctttcaag 5821 agctcggcca tcacgtcaag ctttggataa cgatgaatga gccgtataca aggaatatga 5881 catacagtgc tggccacaac cttctgaagg cccatgccct ggcttggcat gtgtacaatg 5941 aaaagtttag gcatgctcag aatgggaaaa tatccatagc cttgcaggct gattggatag 6001 aacctgcctg ccctttctcc caaaaggaca aagaggtggc cgagagagtt ttggaatttg 6061 acattggctg gctggctgag cccattttcg gctctggaga ttatccatgg gtgatgaggg 6121 actggctgaa ccaaagaaac aattttcttc ttccttattt cactgaagat gaaaaaaagc 6181 taatccaggg tacctttgac tttttggctt taagccatta taccaccatc cttgtagact 6241 cagaaaaaga agatccaata aaatacaatg attacctaga agtgcaagaa atgaccgaca 6301 tcacgtggct caactccccc agtcaggtgg cggtagtgcc ctgggggttg cgcaaagtgc 6361 tgaactggct gaagttcaag tacggagacc tccccatgta cataatatcc aacggaatcg 6421 atgacgggct gcatgctgag gacgaccagc tgagggtgta ttatatgcag aattacataa 6481 acgaagctct caaagcccac atactggatg gtatcaatct ttgcggatac tttgcttatt 6541 cgtttaacga ccgcacagct ccgaggtttg gcctctatcg ttatgctgca gatcagtttg 6601 agcccaaggc atccatgaaa cattacagga aaattattga cagcaatggt ttcccgggcc 6661 cagaaactct ggaaagattt tgtccagaag aattcaccgt gtgtactgag tgcagttttt 6721 ttcacacccg aaagtcttta ctggctttca tagcttttct attttttgct tctattattt 6781 ctctctccct tatattttac tactcgaaga aaggcagaag aagttacaaa TAGGACCCAG 6841 CTTTCTTGTA CAAAGTGGTT GATAGCTTGG ATCCAATCAA CCTCTGGATT ACAAAATTTG 6901 TGAAAGATTG ACTGGTATTC TTAACTATGT TGCTCCTTTT ACGCTATGTG GATACGCTGC 6961 TTTAATGCCT TTGTATCATG CTATTGCTTC CCGTATGGCT TTCATTTTCT CCTCCTTGTA 7021 TAAATCCTGG TTGCTGTCTC TTTATGAGGA GTTGTGGCCC GTTGTCAGGC AACGTGGCGT 7081 GGTGTGCACT GTGTTTGCTG ACGCAACCCC CACTGGTTGG GGCATTGCCA CCACCTGTCA 7141 GCTCCTTTCC GGGACTTTCG CTTTCCCCCT CCCTATTGCC ACGGCGGAAC TCATCGCCGC 7201 CTGCCTTGCC CGCTGCTGGA CAGGGGCTCG GCTGTTGGGC ACTGACAATT CCGTGGTGTT 7261 GTCGGGGAAG CTGACGTCCT TTCCATGGCT GCTCGCCTGT GTTGCCACCT GGATTCTGCG 7321 CGGGACGTCC TTCTGCTACG TCCCTTCGGC CCTCAATCCA GCGGACCTTC CTTCCCGCGG 7381 CCTGCTGCCG GCTCTGCGGC CTCTTCCGCG TCTTCGAGAT CTGCCTCGAC TGTGCCTTCT 7441 AGTTGCCAGC CATCTGTTGT TTGCCCCTCC CCCGTGCCTT CCTTGACCCT GGAAGGTGCC 7501 ACTCCCACTG TCCTTTCCTA ATAAAATGAG GAAATTGCAT CGCATTGTCT GAGTAGGTGT 7561 CATTCTATTC TGGGGGGTGG GGTGGGGCAG GACAGCAAGG GGGAGGATTG GGAAGACAAT 7621 AGCAGGCATG CTGGGGACTC GAGTTAAGGG CGAATTGCCG ATTAGGATCT TCCTAGAGCA 7681 TGGCTACGTA GATAAGTAGC ATGGCGGGTT AATCATTAAC TACAAGGAAC CCCTAGTGAT 7741 GGAGTTGGCC ACTCCCTCTC TGCGCGCTCG CTCGCTCACT GAGGCCGGGC GACCAAAGGT 7801 CGCCCGACGC CCGGGCTTTG CCCGGGCGGC CTCAGTGAGC GAGCGAGCGC GCAGCCTTAA 7861 TTAAGAGCTC ATGGCGCGCC TAGGCCTTGA CGGCCTTCCG CCAATTCGCC CTATAGTGAG 7921 TCGTATTACG TCGCGCTCAC TGGCCGTCGT TTTACAACGT CGTGACTGGG AAAACCCTGG 7981 CGTTACCCAA CTTAATCGCC TTGCAGCACA TCCCCCTTTC GCCAGCTGGC GTAATAGCGA 8041 AGAGGCCCGC ACCGAAACGC CCTTCCCAAC AGTTGCGCAG CCTGAATGGC GAATGGGAGC 8101 GCCCTGTAGC GGCCACTCAA CCCTATCTCG GTCTATTCTT TTGATTTATA AGGGATTTTG 8161 CCGATTTCGG CCTATTGGTT AAAAAATGAG CTGATTTAAC AAAAATTTAA CGCGAATTTT 8221 AACAAAATAT TAACGCTTAC AATTTAG //