LOCUS LCTL-pGEn3-DEST 7551 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 6179..6720 /note="WPRE" misc_recomb 6141..6163 /label=attB2 /note="attB2 site" misc_feature complement(6727..6941) /note="BGH pA" misc_feature complement(7029..7158) /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..6140 /dnas_title="LCTL-pGEn3-DEST" /organism="synthetic construct" /mol_type="other DNA" /db_xref="taxon:32630" /clone="IMAGE:100015340" /clone_lib="NIH_MGC_425" /lab_host="DH10B" /focus /note="Vector: pENTR223.1 without stop codon" source 4593..6137 /organism="Homo sapiens" /mol_type="other DNA" /db_xref="taxon:9606" gene 4593..6137 /dnas_title="LCTL" /gene="LCTL" /gene_synonym="KLG" /gene_synonym="KLPH" /db_xref="GeneID:197021" /db_xref="HGNC:15583" CDS 4593..6137 /dnas_title="lactase-like" /gene="LCTL" /gene_synonym="KLG" /gene_synonym="KLPH" /codon_start=1 /product="lactase-like" /protein_id="AAI48355.1" /db_xref="GI:151556442" /db_xref="GeneID:197021" /db_xref="HGNC:15583" /translation="MKPVWVATLLWMLLLVPRLGAARKGSPEEASFYYGTFPLGFSWG VGSSAYQTEGAWDQDGKGPSIWDVFTHSGKGKVLGNETADVACDGYYKVQEDIILLRE LHVNHYRFSLSWPRLLPTGIRAEQVNKKGIEFYSDLIDALLSSNITPIVTLHHWDLPQ LLQVKYGGWQNVSMANYFRDYANLCFEAFGDRVKHWITFSDPRAMAEKGYETGHHAPG LKLRGTGLYKAAHHIIKAHAKAWHSYNTTWRSKQQGLVGISLNCDWGEPVDISNPKDL EAAERYLQFCLGWFANPIYAGDYPQVMKDYIGRKSAEQGLEMSRLPVFSLQEKSYIKG TSDFLGLGHFTTRYITERNYPSRQGPSYQNDRDLIELVDPNWPDLGSKWLYSVPWGFR RLLNFAQTQYGDPPIYVMENGASQKFHCTQLCDEWRIQYLKGYINEMLKAIKDGANIK GYTSWSLLDKFEWEKGYSDRYGFYYVEFNDRNKPRYPKASVQYYKKIIIANGFPNPRE VESWYLKALETCSINNQMLAAEPLLSHMQMVTEIVVPTVCSLCVLITAVLLMLLLRRQ S" misc_feature 4593..6140 /note="LCTL coding region" misc_feature 6138..6140 /note="linker at the 3' end of the ORF" misc_difference 6139..6140 /note="29 bases at the 3' end do not align to the human genome." vector join(6149..7551,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..6148 /source="LCTL-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 gcccagaaga ggcctccttc tactatggaa 4621 ccttccctct tggcttctcc tggggcgtgg gcagttctgc ctaccagacg gagggcgcct 4681 gggaccagga cgggaaaggg cctagcatct gggacgtctt cacacacagt gggaagggga 4741 aagtgcttgg gaatgagacg gcagatgtag cctgtgacgg ctactacaag gtccaggagg 4801 acatcattct gctgagggaa ctgcacgtca accactaccg attctccctg tcttggcccc 4861 ggctcctgcc cacaggcatc cgagccgagc aggtgaacaa gaagggaatc gaattctaca 4921 gtgatcttat cgatgccctt ctgagcagca acatcactcc catcgtgacc ttgcaccact 4981 gggatctgcc acagctgctc caggtcaaat acggtgggtg gcagaatgtg agcatggcca 5041 actacttcag agactacgcc aacctgtgct ttgaggcctt tggggaccgt gtgaagcact 5101 ggatcacgtt cagtgatcct cgggcaatgg cagaaaaagg ctatgagacg ggccaccatg 5161 cgccgggcct gaagctccgc ggcaccggcc tgtacaaggc agcacaccac atcattaagg 5221 cccacgccaa agcctggcat tcttataaca ccacgtggcg cagcaagcag caaggtctgg 5281 tgggaatttc attgaactgt gactgggggg aacctgtgga cattagtaac cccaaggacc 5341 tagaggctgc cgagagatac ctacagttct gtctgggctg gtttgccaac cccatttatg 5401 ccggtgacta cccccaagtc atgaaggact acattggaag aaagagtgca gagcaaggcc 5461 tggagatgtc gaggttaccg gtgttctcac tccaggagaa gagctacatt aaaggcacat 5521 ccgatttctt gggattaggt cattttacta ctcggtacat cacggaaagg aactacccct 5581 cccgccaggg gcccagctac cagaacgatc gtgacttgat agagctggtt gacccaaact 5641 ggccagatct ggggtctaaa tggctatatt ctgtgccatg gggatttagg aggctcctta 5701 actttgctca gactcaatac ggtgatcctc ccatatatgt gatggaaaat ggagcatctc 5761 aaaaattcca ctgtactcaa ttatgtgatg agtggagaat tcaatacctt aaaggataca 5821 taaatgaaat gctaaaagct ataaaagatg gtgctaatat aaaggggtat acttcctggt 5881 ctctgttgga taagtttgaa tgggagaaag gatactcaga tagatatgga ttctactatg 5941 ttgaatttaa cgacagaaat aagcctcgct atccaaaggc ttcagttcaa tattacaaga 6001 agattatcat tgccaatggg tttcccaatc caagagaggt ggaaagttgg tacctcaaag 6061 ctttggaaac ttgctctatc aacaatcaga tgcttgctgc agagcccttg ctaagtcaca 6121 tgcaaatggt tacggagtga CCAGCTTTCT TGTACAAAGT GGTTGATAGC TTGGATCCAA 6181 TCAACCTCTG GATTACAAAA TTTGTGAAAG ATTGACTGGT ATTCTTAACT ATGTTGCTCC 6241 TTTTACGCTA TGTGGATACG CTGCTTTAAT GCCTTTGTAT CATGCTATTG CTTCCCGTAT 6301 GGCTTTCATT TTCTCCTCCT TGTATAAATC CTGGTTGCTG TCTCTTTATG AGGAGTTGTG 6361 GCCCGTTGTC AGGCAACGTG GCGTGGTGTG CACTGTGTTT GCTGACGCAA CCCCCACTGG 6421 TTGGGGCATT GCCACCACCT GTCAGCTCCT TTCCGGGACT TTCGCTTTCC CCCTCCCTAT 6481 TGCCACGGCG GAACTCATCG CCGCCTGCCT TGCCCGCTGC TGGACAGGGG CTCGGCTGTT 6541 GGGCACTGAC AATTCCGTGG TGTTGTCGGG GAAGCTGACG TCCTTTCCAT GGCTGCTCGC 6601 CTGTGTTGCC ACCTGGATTC TGCGCGGGAC GTCCTTCTGC TACGTCCCTT CGGCCCTCAA 6661 TCCAGCGGAC CTTCCTTCCC GCGGCCTGCT GCCGGCTCTG CGGCCTCTTC CGCGTCTTCG 6721 AGATCTGCCT CGACTGTGCC TTCTAGTTGC CAGCCATCTG TTGTTTGCCC CTCCCCCGTG 6781 CCTTCCTTGA CCCTGGAAGG TGCCACTCCC ACTGTCCTTT CCTAATAAAA TGAGGAAATT 6841 GCATCGCATT GTCTGAGTAG GTGTCATTCT ATTCTGGGGG GTGGGGTGGG GCAGGACAGC 6901 AAGGGGGAGG ATTGGGAAGA CAATAGCAGG CATGCTGGGG ACTCGAGTTA AGGGCGAATT 6961 GCCGATTAGG ATCTTCCTAG AGCATGGCTA CGTAGATAAG TAGCATGGCG GGTTAATCAT 7021 TAACTACAAG GAACCCCTAG TGATGGAGTT GGCCACTCCC TCTCTGCGCG CTCGCTCGCT 7081 CACTGAGGCC GGGCGACCAA AGGTCGCCCG ACGCCCGGGC TTTGCCCGGG CGGCCTCAGT 7141 GAGCGAGCGA GCGCGCAGCC TTAATTAAGA GCTCATGGCG CGCCTAGGCC TTGACGGCCT 7201 TCCGCCAATT CGCCCTATAG TGAGTCGTAT TACGTCGCGC TCACTGGCCG TCGTTTTACA 7261 ACGTCGTGAC TGGGAAAACC CTGGCGTTAC CCAACTTAAT CGCCTTGCAG CACATCCCCC 7321 TTTCGCCAGC TGGCGTAATA GCGAAGAGGC CCGCACCGAA ACGCCCTTCC CAACAGTTGC 7381 GCAGCCTGAA TGGCGAATGG GAGCGCCCTG TAGCGGCCAC TCAACCCTAT CTCGGTCTAT 7441 TCTTTTGATT TATAAGGGAT TTTGCCGATT TCGGCCTATT GGTTAAAAAA TGAGCTGATT 7501 TAACAAAAAT TTAACGCGAA TTTTAACAAA ATATTAACGC TTACAATTTA G //