LOCUS GCS1-pGEn3-DEST 8340 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(4541..4569) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 6968..7509 /note="WPRE" misc_recomb 6930..6952 /label=attB2 /note="attB2 site" misc_feature complement(7516..7730) /note="BGH pA" misc_feature complement(7818..7947) /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..6929 /dnas_title="GCS1-pGEn3-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:34915 IMAGE:5110525" /tissue_type="Cervix, carcinoma" /clone_lib="NIH_MGC_12" /lab_host="DH10B" /note="Vector: pCMV-SPORT6" gene 4593..6929 /dnas_title="GCS1" /gene="GCS1" /db_xref="GeneID:7841" /db_xref="MIM:601336" CDS 4593..6929 /dnas_title="glucosidase I" /gene="GCS1" /codon_start=1 /product="glucosidase I" /protein_id="AAH28337.1" /db_xref="GI:62020522" /db_xref="GeneID:7841" /db_xref="MIM:601336" /translation="MARGERRRRAVPAEGVRTAERAARGGPGRRDGRGGGPRSTAGGV ALAVVVLSLALGMSGRWVLAWYRARRAVTLHSAPPVLPADSSSPAVAPDLFWGTYRPH VYFGMKTRSPKPLLTGLMWAQQGTTPGTPKLRHTCEQGDGVGPYGWEFHDGLSFGRQH IQDGALRLTTEFVKRPGGQHGGDWSWRVTVEPQDSGTSALPLVSLFFYVVTDGKEVLL PEVGAKGQLKFISGHTSELGNFRFTLLPPTSPGDTAPKYGSYNVFWTSNPGLPLLTEM VKSRLNSWFQHRPPGASPERYLGLPGSLKWEDRGPSGQGQGQFLIQQVTLKIPISIEF VFESGSAQAGGNQALPRLAGSLLTQALESHAEGFRERFEKTFQLKEKGLSSGEQVLGQ AALSGLLGGIGYFYGQGLVLPDIGVEGSEQKVDPALFPPVPLFTAVPSRSFFPRGFLW DEGFHQLVVQRWDPSLTREALGHWLGLLNADGWIGREQILGDEARARVPPEFLVQRAV HANPPTLLLPVAHMLEVGDPDDLAFLRKALPRLHAWFSWLHQSQAGPLPLSYRWRGRD PALPTLLNPKTLPSGLDDYPRASHPSVTERHLDLRCWVALGARVLTRLAEHLGEAEVA AELGPLAASLEAAESLDELHWAPELGVFADFGNHTKAVQLKPRPPQGLVRVVGRPQPQ LQYVDALGYVSLFPLLLRLLDPTSSRLGPLLDILADSRHLWSPFGLRSLAASSSFYGQ RNSEHDPPYWRGAVWLNVNYLALGALHHYGHLEGPHQARAAKLHGELRANVVGNVWRQ YQATGFLWEQYSDRDGRGMGCRPFHGWTSLVLLAMAEDY" misc_feature 4593..6929 /note="GCS1 coding region" misc_difference 5130..5130 /dnas_title="GCS1" /gene="GCS1" /note="'A' in cDNA is 'G' in the human genome; amino acid difference: 'N' in cDNA, 'D' in the human genome. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." misc_difference 5292..5292 /dnas_title="GCS1" /gene="GCS1" /note="'T' in cDNA is 'C' in the human genome; amino acid difference: 'S' in cDNA, 'P' in the human genome." vector join(6938..8340,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..6937 /source="GCS1-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 gccgctgggt gctggcgtgg taccgtgcgc 4621 ggcgggcggt cacgctgcac tccgcgcctc ctgtgttgcc tgccgactcc tccagccccg 4681 ccgtggcccc ggacctcttc tggggaacct accgccctca cgtctacttc ggcatgaaga 4741 cccgcagccc gaagcccctc ctcaccggac tgatgtgggc gcagcagggc accaccccgg 4801 ggactcctaa gctcaggcac acgtgtgagc agggggacgg tgtgggtccc tatggctggg 4861 agttccacga cggcctctcc ttcgggcgcc aacacatcca ggatggggcc ttaaggctca 4921 ccactgagtt cgtcaagagg cctgggggtc agcacggagg ggactggagc tggagagtga 4981 ctgtagagcc tcaggactca ggtacttctg ccctcccttt ggtctccctg ttcttctatg 5041 tggtgacaga tggcaaggaa gtcctactac cagaggttgg ggccaagggg cagttgaagt 5101 ttatcagtgg gcacaccagt gaacttggta acttccgctt tacacttttg ccaccaacca 5161 gtccagggga tacagccccc aagtatggca gctacaatgt cttctggacc tccaacccag 5221 gactgcccct gctgacagag atggtaaaga gtcgcctaaa tagctggttt cagcatcggc 5281 ccccaggggc ctcccctgaa cgctacctcg gcttgccagg atccctgaag tgggaggaca 5341 gaggtccaag tgggcaaggg caggggcagt tcttgataca gcaggtgacc ctgaaaattc 5401 ccatttccat agagtttgtg tttgaatcag gcagtgccca ggcaggagga aatcaagccc 5461 tgccaagact ggcaggcagt ctactgaccc aggccctgga gagccatgct gaaggcttta 5521 gagagcgctt tgagaagacc ttccagctga aggagaaggg cctgagctct ggcgagcagg 5581 ttttgggtca ggctgccctc agcggcctcc ttggtggaat tggctacttc tacggacaag 5641 ggctggtatt gccagacatc ggggtggaag ggtctgagca gaaggtggac ccagccctct 5701 ttccacccgt acctcttttt acagcagtgc cctcccggtc attcttccca cgaggcttcc 5761 tttgggatga aggctttcac cagctggtgg ttcagcggtg ggatccctcc ctcacccggg 5821 aagcccttgg ccactggctg gggctgctaa atgctgatgg ctggattggg agggagcaga 5881 tactggggga tgaggcccga gcccgggtgc ctccagaatt cctagtacaa cgagcagtcc 5941 acgccaaccc cccaacccta cttttgcctg tagcccatat gctagaggtt ggtgaccctg 6001 acgacttggc tttcctccga aaggccttgc cccgcctgca tgcctggttt tcctggctcc 6061 atcagagcca ggcaggccca ctgccactat cttaccgctg gcggggacgg gaccctgcct 6121 taccaacctt actgaacccc aagaccctac cctctgggct ggatgactac ccccgggctt 6181 cacacccttc agtaaccgag cggcacctgg acctgcgatg ttgggtggca ctgggtgccc 6241 gtgtgctgac gcggctggca gagcatctgg gtgaggctga ggtagctgct gagctgggcc 6301 cactggctgc ctcactggag gcagcagaga gcctggatga gctgcactgg gccccagagc 6361 taggagtctt tgcagacttt gggaaccaca caaaagcagt acagctgaag cccaggcccc 6421 ctcaggggct cgttcgggtg gtgggtcggc cccaacctca actgcagtat gtagatgctc 6481 ttggctatgt cagtcttttt cccttgctgc tgcgactgct ggaccccacc tcatcccgcc 6541 ttgggcccct gctggacatt ctagccgaca gccgccatct ctggagcccc tttggtttac 6601 gctcccttgc agcctccagc tccttttatg gccagcgcaa ttcagagcat gatcccccct 6661 actggcgggg tgctgtgtgg ctcaatgtca actacctggc tttgggagca ctccaccact 6721 atgggcatct ggagggtcct caccaggctc gggctgccaa actccacggt gagctccgtg 6781 ccaacgtggt aggcaatgta tggcgccagt accaggctac aggctttctt tgggagcagt 6841 acagtgaccg cgatgggcga ggcatgggct gccgcccttt ccacggctgg accagccttg 6901 tcttactggc catggctgaa gactactgaC CAGCTTTCTT GTACAAAGTG GTTGATAGCT 6961 TGGATCCAAT CAACCTCTGG ATTACAAAAT TTGTGAAAGA TTGACTGGTA TTCTTAACTA 7021 TGTTGCTCCT TTTACGCTAT GTGGATACGC TGCTTTAATG CCTTTGTATC ATGCTATTGC 7081 TTCCCGTATG GCTTTCATTT TCTCCTCCTT GTATAAATCC TGGTTGCTGT CTCTTTATGA 7141 GGAGTTGTGG CCCGTTGTCA GGCAACGTGG CGTGGTGTGC ACTGTGTTTG CTGACGCAAC 7201 CCCCACTGGT TGGGGCATTG CCACCACCTG TCAGCTCCTT TCCGGGACTT TCGCTTTCCC 7261 CCTCCCTATT GCCACGGCGG AACTCATCGC CGCCTGCCTT GCCCGCTGCT GGACAGGGGC 7321 TCGGCTGTTG GGCACTGACA ATTCCGTGGT GTTGTCGGGG AAGCTGACGT CCTTTCCATG 7381 GCTGCTCGCC TGTGTTGCCA CCTGGATTCT GCGCGGGACG TCCTTCTGCT ACGTCCCTTC 7441 GGCCCTCAAT CCAGCGGACC TTCCTTCCCG CGGCCTGCTG CCGGCTCTGC GGCCTCTTCC 7501 GCGTCTTCGA GATCTGCCTC GACTGTGCCT TCTAGTTGCC AGCCATCTGT TGTTTGCCCC 7561 TCCCCCGTGC CTTCCTTGAC CCTGGAAGGT GCCACTCCCA CTGTCCTTTC CTAATAAAAT 7621 GAGGAAATTG CATCGCATTG TCTGAGTAGG TGTCATTCTA TTCTGGGGGG TGGGGTGGGG 7681 CAGGACAGCA AGGGGGAGGA TTGGGAAGAC AATAGCAGGC ATGCTGGGGA CTCGAGTTAA 7741 GGGCGAATTG CCGATTAGGA TCTTCCTAGA GCATGGCTAC GTAGATAAGT AGCATGGCGG 7801 GTTAATCATT AACTACAAGG AACCCCTAGT GATGGAGTTG GCCACTCCCT CTCTGCGCGC 7861 TCGCTCGCTC ACTGAGGCCG GGCGACCAAA GGTCGCCCGA CGCCCGGGCT TTGCCCGGGC 7921 GGCCTCAGTG AGCGAGCGAG CGCGCAGCCT TAATTAAGAG CTCATGGCGC GCCTAGGCCT 7981 TGACGGCCTT CCGCCAATTC GCCCTATAGT GAGTCGTATT ACGTCGCGCT CACTGGCCGT 8041 CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC 8101 ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGAAA CGCCCTTCCC 8161 AACAGTTGCG CAGCCTGAAT GGCGAATGGG AGCGCCCTGT AGCGGCCACT CAACCCTATC 8221 TCGGTCTATT CTTTTGATTT ATAAGGGATT TTGCCGATTT CGGCCTATTG GTTAAAAAAT 8281 GAGCTGATTT AACAAAAATT TAACGCGAAT TTTAACAAAA TATTAACGCT TACAATTTAG //