LOCUS EDEM1-pGEn2-DEST 7200 bp DNA circular 25-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 5828..6369 /note="WPRE" misc_recomb 5790..5812 /label=attB2 /note="attB2 site" misc_feature complement(6376..6590) /note="BGH pA" misc_feature complement(6678..6807) /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..5789 /dnas_title="EDEM1-pGEn2-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:29498 IMAGE:4863779" /tissue_type="Skin, melanotic melanoma, high MDR." /clone_lib="NIH_MGC_49" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 3891..5789 /dnas_title="EDEM1" /gene="EDEM1" /gene_synonym="EDEM" /gene_synonym="KIAA0212" /db_xref="GeneID:9695" /db_xref="HGNC:18967" /db_xref="MIM:607673" CDS 3891..5789 /dnas_title="ER degradation enhancer, mannosidase alpha-like 1" /gene="EDEM1" /gene_synonym="EDEM" /gene_synonym="KIAA0212" /codon_start=1 /product="ER degradation enhancer, mannosidase alpha-like 1" /protein_id="AAH19088.1" /db_xref="GI:17512212" /db_xref="GeneID:9695" /db_xref="HGNC:18967" /db_xref="MIM:607673" /translation="MQWRALVLGLVLLRLGLHGVLWLVFGLGPSMGFYQRFPLSFGFQ RLRSPDGPASPTSGPVGRPGGVSGPSWLQPPGTGAAQSPRKAPRRPGPGMCGPANWGY VLGGRGRGPDEYEKRYSGAFPPQLRAQMRDLARGMFVFGYDNYMAHAFPQDELNPIHC RGRGPDRGDPSNLNINDVLGNYSLTLVDALDTLAIMGNSSEFQKAVKLVINTVSFDKD STVQVFEATIRVLGSLLSAHRIITDSKQPFGDMTIKDYDNELLYMAHDLAVRLLPAFE NTKTGIPYPRVNLKTGVPPDTNNETCTAGAGSLLVEFGILSRLLGDSTFEWVARRAVK ALWNLRSNDTGLLGNVVNIQTGHWVGKQSGLGAGLDSFYEYLLKSYILFGEKEDLEMF NAAYQSIQNYLRRGREACNEGEGDPPLYVNVNMFSGQLMNTWIDSLQAFFPGLQVLIG DVEDAICLHAFYYAIWKRYGALPERYNWQLQAPDVLFYPLRPELVESTYLLYQATKNP FYLHVGMDILQSLEKYTKVKCGYATLHHVIDKSTEDRMESFFLSETCKYLYLLFDEDN PVHKSGTRYMFTTEGHIVSVDEHLRELPWKEFFSEEGGQDQGGKSVHRPKPHELKVIN SSSNCNRVPDERRYSLPLKSIYMRQIDQMVGLI" misc_feature 3891..5789 /note="EDEM1 coding region" misc_difference 4430..4430 /dnas_title="EDEM1" /gene="EDEM1" /gene_synonym="EDEM" /gene_synonym="KIAA0212" /note="'A' in cDNA is 'C' in the human genome; no amino acid change. The chimpanzee genome agrees with the cDNA sequence, suggesting that this difference is unlikely to be due to an artifact." vector join(5798..7200,1..3857) /source="pGEn2-DEST" /type="Destination Vector" clone 3858..5797 /source="EDEM1-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 gggctggggc 3901 ccagcatggg cttctaccag cgctttccgc tcagcttcgg cttccagcgt ctgaggagcc 3961 ccgacggccc cgcgtcgccc acctcggggc ccgtgggccg gcctgggggg gtatccgggc 4021 cgtcgtggct gcagccgccg gggaccgggg cagcgcagag cccgcgcaag gctccgcggc 4081 gtcctgggcc ggggatgtgc ggcccagcca actggggcta cgtgctgggc ggccggggcc 4141 gcggcccgga cgagtacgag aagcgctaca gcggcgcctt ccctccgcag ctgcgtgccc 4201 agatgcgcga cctggcacgg ggcatgttcg tctttggcta cgacaactac atggctcacg 4261 ccttccccca ggacgagctc aaccccatcc actgccgcgg ccgtgggccc gaccgcgggg 4321 acccttcaaa tctgaacatc aatgatgtac tagggaacta ctcattgact cttgttgatg 4381 cattggatac acttgcaata atgggaaatt catccgagtt ccagaaagca gtcaagttag 4441 tgatcaacac agtttcattt gacaaagatt ccaccgtcca agtctttgag gccacgataa 4501 gggtcctggg aagcctcctt tctgctcaca gaataataac tgactccaag cagccctttg 4561 gtgacatgac aattaaggac tatgataatg agttgttata catggcccat gacctggcgg 4621 tgcggctcct ccctgctttt gaaaacacca agacagggat tccatatcct cgggtgaatc 4681 taaagacagg agttcctcct gacaccaata atgagacatg cacagcggga gccggttccc 4741 tcctggtgga atttgggatt ctgagtcgac tcctggggga ctccacattt gagtgggtgg 4801 ccagacgagc agtgaaagcc ctttggaacc tccggagcaa tgatacagga ttactaggca 4861 atgtcgtgaa cattcagacg ggccactggg ttggaaagca gagtggcctg ggtgccgggc 4921 tggactcctt ctatgaatac ctcttgaaat cttacattct ctttggagaa aaagaagacc 4981 tagaaatgtt taatgctgca tatcagagta ttcagaacta cttaagaaga gggcgggaag 5041 cctgcaatga aggagaagga gaccctccac tctatgtcaa cgtgaacatg ttcagtgggc 5101 agctgatgaa cacctggatt gactctctgc aggccttttt ccctggactg caggtgctga 5161 taggagatgt ggaagatgcc atctgccttc atgccttcta ctatgccata tggaaacgat 5221 atggtgccct ccctgagaga tataactggc agctgcaggc ccctgacgtt ctcttctacc 5281 cactgagacc agagttagtg gaatccacat atctcctcta ccaggcaacc aagaatccct 5341 tctacctcca tgtaggaatg gatattctgc agagtctgga aaagtacaca aaagtcaagt 5401 gtgggtacgc cacgctgcat cacgtcattg acaagtccac agaagaccgg atggagagct 5461 tctttctcag tgagacctgt aaatatttgt atctgctgtt tgatgaagac aatccagtac 5521 acaagtctgg aaccagatac atgttcacaa cagagggaca cattgtatct gtggatgagc 5581 atcttcggga attgccatgg aaggaattct tctctgaaga gggagggcag gaccaagggg 5641 gaaagtctgt gcacaggccg aaacctcatg agttaaaagt catcaactcc agctccaact 5701 gcaatcgtgt acctgatgag aggaggtact ccctgccctt aaagagcatc tacatgcgac 5761 agattgacca gatggttggt ttgatttgaC CAGCTTTCTT GTACAAAGTG GTTGATAGCT 5821 TGGATCCAAT CAACCTCTGG ATTACAAAAT TTGTGAAAGA TTGACTGGTA TTCTTAACTA 5881 TGTTGCTCCT TTTACGCTAT GTGGATACGC TGCTTTAATG CCTTTGTATC ATGCTATTGC 5941 TTCCCGTATG GCTTTCATTT TCTCCTCCTT GTATAAATCC TGGTTGCTGT CTCTTTATGA 6001 GGAGTTGTGG CCCGTTGTCA GGCAACGTGG CGTGGTGTGC ACTGTGTTTG CTGACGCAAC 6061 CCCCACTGGT TGGGGCATTG CCACCACCTG TCAGCTCCTT TCCGGGACTT TCGCTTTCCC 6121 CCTCCCTATT GCCACGGCGG AACTCATCGC CGCCTGCCTT GCCCGCTGCT GGACAGGGGC 6181 TCGGCTGTTG GGCACTGACA ATTCCGTGGT GTTGTCGGGG AAGCTGACGT CCTTTCCATG 6241 GCTGCTCGCC TGTGTTGCCA CCTGGATTCT GCGCGGGACG TCCTTCTGCT ACGTCCCTTC 6301 GGCCCTCAAT CCAGCGGACC TTCCTTCCCG CGGCCTGCTG CCGGCTCTGC GGCCTCTTCC 6361 GCGTCTTCGA GATCTGCCTC GACTGTGCCT TCTAGTTGCC AGCCATCTGT TGTTTGCCCC 6421 TCCCCCGTGC CTTCCTTGAC CCTGGAAGGT GCCACTCCCA CTGTCCTTTC CTAATAAAAT 6481 GAGGAAATTG CATCGCATTG TCTGAGTAGG TGTCATTCTA TTCTGGGGGG TGGGGTGGGG 6541 CAGGACAGCA AGGGGGAGGA TTGGGAAGAC AATAGCAGGC ATGCTGGGGA CTCGAGTTAA 6601 GGGCGAATTG CCGATTAGGA TCTTCCTAGA GCATGGCTAC GTAGATAAGT AGCATGGCGG 6661 GTTAATCATT AACTACAAGG AACCCCTAGT GATGGAGTTG GCCACTCCCT CTCTGCGCGC 6721 TCGCTCGCTC ACTGAGGCCG GGCGACCAAA GGTCGCCCGA CGCCCGGGCT TTGCCCGGGC 6781 GGCCTCAGTG AGCGAGCGAG CGCGCAGCCT TAATTAAGAG CTCATGGCGC GCCTAGGCCT 6841 TGACGGCCTT CCGCCAATTC GCCCTATAGT GAGTCGTATT ACGTCGCGCT CACTGGCCGT 6901 CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC 6961 ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGAAA CGCCCTTCCC 7021 AACAGTTGCG CAGCCTGAAT GGCGAATGGG AGCGCCCTGT AGCGGCCACT CAACCCTATC 7081 TCGGTCTATT CTTTTGATTT ATAAGGGATT TTGCCGATTT CGGCCTATTG GTTAAAAAAT 7141 GAGCTGATTT AACAAAAATT TAACGCGAAT TTTAACAAAA TATTAACGCT TACAATTTAG //