LOCUS GAA-pGEn3-DEST 8781 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 7409..7950 /note="WPRE" misc_recomb 7371..7393 /label=attB2 /note="attB2 site" misc_feature complement(7957..8171) /note="BGH pA" misc_feature complement(8259..8388) /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..7370 /dnas_title="GAA-pGEn3-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:48746 IMAGE:5739991" /tissue_type="Duodenum, adenocarcinoma" /clone_lib="NIH_MGC_88" /lab_host="DH10B" /note="Vector: pCMV-SPORT6" gene 4593..7370 /dnas_title="GAA" /gene="GAA" /gene_synonym="LYAG" /db_xref="GeneID:2548" /db_xref="HGNC:4065" /db_xref="MIM:606800" CDS 4593..7370 /dnas_title="glucosidase, alpha; acid" /gene="GAA" /gene_synonym="LYAG" /codon_start=1 /product="glucosidase, alpha; acid" /protein_id="AAH40431.1" /db_xref="GI:26251857" /db_xref="GeneID:2548" /db_xref="HGNC:4065" /db_xref="MIM:606800" /translation="MGVRHPPCSHRLLAVCALVSLATAALLGHILLHDFLLVPRELSG SSPVLEETHPAHQQGASRPGPRDAQAHPGRPRAVPTQCDVPPNSRFDCAPDKAITQEQ CEARGCCYIPAKQGLQGAQMGQPWCFFPPSYPSYKLENLSSSEMGYTATLTRTTPTFF PKDILTLRLDVMMETENRLHFTIKDPANRRYEVPLETPHVHSRAPSPLYSVEFSEEPF GVIVRRQLDGRVLLNTTVAPLFFADQFLQLSTSLPSQYITGLAEHLSPLMLSTSWTRI TLWNRDLAPTPGANLYGSHPFYLALEDGGSAHGVFLLNSNAMDVVLQPSPALSWRSTG GILDVYIFLGPEPKSVVQQYLDVVGYPFMPPYWGLGFHLCRWGYSSTAITRQVVENMT RAHFPLDVQWNDLDYMDSRRDFTFNKDGFRDFPAMVQELHQGGRRYMMIVDPAISSSG PAGSYRPYDEGLRRGVFITNETGQPLIGKVWPGSTAFPDFTNPTALAWWEDMVAEFHD QVPFDGMWIDMNEPSNFIRGSEDGCPNNELENPPYVPGVVGGTLQAATICASSHQFLS THYNLHNLYGLTEAIASHRALVKARGTRPFVISRSTFAGHGRYAGHWTGDVWSSWEQL ASSVPEILQFNLLGVPLVGADVCGFLGNTSEELCVRWTQLGAFYPFMRNHNSLLSLPQ EPYSFSEPAQQAMRKALTLRYALLPHLYTLFHQAHVAGETVARPLFLEFPKDSSTWTV DHQLLWGEALLITPVLQAGKAEVTGYFPLGTWYDLQTVPVEALGSLPPPPAAPREPAI HSEGQWVTLPAPLDTINVHLRAGYIIPLQGPGLTTTESRQQPMALAVALTKGGEARGE LFWDDGESLEVLERGAYTQVIFLARNNTIVNELVRVTSEGAGLQLQKVTVLGVATAPQ QVLSNGVPVSNFTYSPDTKVLDICVSLLMGEQFLVSWC" misc_difference 4694..4694 /dnas_title="GAA" /gene="GAA" /gene_synonym="LYAG" /note="'T' in cDNA is 'C' in the human genome; no amino acid change. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." misc_feature 4593..7370 /note="GAA coding region" misc_difference 7082..7082 /dnas_title="GAA" /gene="GAA" /gene_synonym="LYAG" /note="'T' in cDNA is 'G' in the human genome; no amino acid change. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." vector join(7379..8781,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..7378 /source="GAA-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 gcgggcacat cctactccat gatttcctgc 4621 tggttccccg agagctgagt ggctcctccc cagtcctgga ggagactcac ccagctcacc 4681 agcagggagc cagtagacca gggccccggg atgcccaggc acaccccggc cgtcccagag 4741 cagtgcccac acagtgcgac gtccccccca acagccgctt cgattgcgcc cctgacaagg 4801 ccatcaccca ggaacagtgc gaggcccgcg gctgttgcta catccctgca aagcaggggc 4861 tgcagggagc ccagatgggg cagccctggt gcttcttccc acccagctac cccagctaca 4921 agctggagaa cctgagctcc tctgaaatgg gctacacggc caccctgacc cgtaccaccc 4981 ccaccttctt ccccaaggac atcctgaccc tgcggctgga cgtgatgatg gagactgaga 5041 accgcctcca cttcacgatc aaagatccag ctaacaggcg ctacgaggtg cccttggaga 5101 ccccgcatgt ccacagccgg gcaccgtccc cactctacag cgtggagttc tccgaggagc 5161 ccttcggggt gatcgtgcgc cggcagctgg acggccgcgt gctgctgaac acgacggtgg 5221 cgcccctgtt ctttgcggac cagttccttc agctgtccac ctcgctgccc tcgcagtata 5281 tcacaggcct cgccgagcac ctcagtcccc tgatgctcag caccagctgg accaggatca 5341 ccctgtggaa ccgggacctt gcgcccacgc ccggtgcgaa cctctacggg tctcaccctt 5401 tctacctggc gctggaggac ggcgggtcgg cacacggggt gttcctgcta aacagcaatg 5461 ccatggatgt ggtcctgcag ccgagccctg cccttagctg gaggtcgaca ggtgggatcc 5521 tggatgtcta catcttcctg ggcccagagc ccaagagcgt ggtgcagcag tacctggacg 5581 ttgtgggata cccgttcatg ccgccatact ggggcctggg cttccacctg tgccgctggg 5641 gctactcctc caccgctatc acccgccagg tggtggagaa catgaccagg gcccacttcc 5701 ccctggacgt ccagtggaac gacctggact acatggactc ccggagggac ttcacgttca 5761 acaaggatgg cttccgggac ttcccggcca tggtgcagga gctgcaccag ggcggccggc 5821 gctacatgat gatcgtggat cctgccatca gcagctcggg ccctgccggg agctacaggc 5881 cctacgacga gggtctgcgg aggggggttt tcatcaccaa cgagaccggc cagccgctga 5941 ttgggaaggt atggcccggg tccactgcct tccccgactt caccaacccc acagccctgg 6001 cctggtggga ggacatggtg gctgagttcc atgaccaggt gcccttcgac ggcatgtgga 6061 ttgacatgaa cgagccttcc aacttcatca ggggctctga ggacggctgc cccaacaatg 6121 agctggagaa cccaccctac gtgcctgggg tggttggggg gaccctccag gcggccacca 6181 tctgtgcctc cagccaccag tttctctcca cacactacaa cctgcacaac ctctacggcc 6241 tgaccgaagc catcgcctcc cacagggcgc tggtgaaggc tcgggggaca cgcccatttg 6301 tgatctcccg ctcgaccttt gctggccacg gccgatacgc cggccactgg acgggggacg 6361 tgtggagctc ctgggagcag ctcgcctcct ccgtgccaga aatcctgcag tttaacctgc 6421 tgggggtgcc tctggtcggg gccgacgtct gcggcttcct gggcaacacc tcagaggagc 6481 tgtgtgtgcg ctggacccag ctgggggcct tctacccctt catgcggaac cacaacagcc 6541 tgctcagtct gccccaggag ccgtacagct tcagcgagcc ggcccagcag gccatgagga 6601 aggccctcac cctgcgctac gcactcctcc cccacctcta cacactgttc caccaggccc 6661 acgtcgcggg ggagaccgtg gcccggcccc tcttcctgga gttccccaag gactctagca 6721 cctggactgt ggaccaccag ctcctgtggg gggaggccct gctcatcacc ccagtgctcc 6781 aggccgggaa ggccgaagtg actggctact tccccttggg cacatggtac gacctgcaga 6841 cggtgccagt agaggccctt ggcagcctcc cacccccacc tgcagctccc cgtgagccag 6901 ccatccacag cgaggggcag tgggtgacgc tgccggcccc cctggacacc atcaacgtcc 6961 acctccgggc tgggtacatc atccccctgc agggccctgg cctcacaacc acagagtccc 7021 gccagcagcc catggccctg gctgtggccc tgaccaaggg tggggaggcc cgaggggagc 7081 ttttctggga cgatggagag agcctggaag tgctggagcg aggggcctac acacaggtca 7141 tcttcctggc caggaataac acgatcgtga atgagctggt acgtgtgacc agtgagggag 7201 ctggcctgca gctgcagaag gtgactgtcc tgggcgtggc cacggcgccc cagcaggtcc 7261 tctccaacgg tgtccctgtc tccaacttca cctacagccc cgacaccaag gtcctggaca 7321 tctgtgtctc gctgttgatg ggagagcagt ttctcgtcag ctggtgttag CCAGCTTTCT 7381 TGTACAAAGT GGTTGATAGC TTGGATCCAA TCAACCTCTG GATTACAAAA TTTGTGAAAG 7441 ATTGACTGGT ATTCTTAACT ATGTTGCTCC TTTTACGCTA TGTGGATACG CTGCTTTAAT 7501 GCCTTTGTAT CATGCTATTG CTTCCCGTAT GGCTTTCATT TTCTCCTCCT TGTATAAATC 7561 CTGGTTGCTG TCTCTTTATG AGGAGTTGTG GCCCGTTGTC AGGCAACGTG GCGTGGTGTG 7621 CACTGTGTTT GCTGACGCAA CCCCCACTGG TTGGGGCATT GCCACCACCT GTCAGCTCCT 7681 TTCCGGGACT TTCGCTTTCC CCCTCCCTAT TGCCACGGCG GAACTCATCG CCGCCTGCCT 7741 TGCCCGCTGC TGGACAGGGG CTCGGCTGTT GGGCACTGAC AATTCCGTGG TGTTGTCGGG 7801 GAAGCTGACG TCCTTTCCAT GGCTGCTCGC CTGTGTTGCC ACCTGGATTC TGCGCGGGAC 7861 GTCCTTCTGC TACGTCCCTT CGGCCCTCAA TCCAGCGGAC CTTCCTTCCC GCGGCCTGCT 7921 GCCGGCTCTG CGGCCTCTTC CGCGTCTTCG AGATCTGCCT CGACTGTGCC TTCTAGTTGC 7981 CAGCCATCTG TTGTTTGCCC CTCCCCCGTG CCTTCCTTGA CCCTGGAAGG TGCCACTCCC 8041 ACTGTCCTTT CCTAATAAAA TGAGGAAATT GCATCGCATT GTCTGAGTAG GTGTCATTCT 8101 ATTCTGGGGG GTGGGGTGGG GCAGGACAGC AAGGGGGAGG ATTGGGAAGA CAATAGCAGG 8161 CATGCTGGGG ACTCGAGTTA AGGGCGAATT GCCGATTAGG ATCTTCCTAG AGCATGGCTA 8221 CGTAGATAAG TAGCATGGCG GGTTAATCAT TAACTACAAG GAACCCCTAG TGATGGAGTT 8281 GGCCACTCCC TCTCTGCGCG CTCGCTCGCT CACTGAGGCC GGGCGACCAA AGGTCGCCCG 8341 ACGCCCGGGC TTTGCCCGGG CGGCCTCAGT GAGCGAGCGA GCGCGCAGCC TTAATTAAGA 8401 GCTCATGGCG CGCCTAGGCC TTGACGGCCT TCCGCCAATT CGCCCTATAG TGAGTCGTAT 8461 TACGTCGCGC TCACTGGCCG TCGTTTTACA ACGTCGTGAC TGGGAAAACC CTGGCGTTAC 8521 CCAACTTAAT CGCCTTGCAG CACATCCCCC TTTCGCCAGC TGGCGTAATA GCGAAGAGGC 8581 CCGCACCGAA ACGCCCTTCC CAACAGTTGC GCAGCCTGAA TGGCGAATGG GAGCGCCCTG 8641 TAGCGGCCAC TCAACCCTAT CTCGGTCTAT TCTTTTGATT TATAAGGGAT TTTGCCGATT 8701 TCGGCCTATT GGTTAAAAAA TGAGCTGATT TAACAAAAAT TTAACGCGAA TTTTAACAAA 8761 ATATTAACGC TTACAATTTA G //