LOCUS GAA-pGEn1-DEST 7354 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_feature 2977..3054 /note="TCM Sig Seq" misc_recomb complement(3114..3142) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 3055..3078 /note="8xHis Tag" misc_feature 3085..3108 /note="StrepTag" misc_feature 5982..6523 /note="WPRE" misc_recomb 5944..5966 /label=attB2 /note="attB2 site" misc_feature complement(6530..6744) /note="BGH pA" misc_feature complement(6832..6961) /note="3'-ITR" misc_feature 3143..3165 /note="TEV site" source 3166..5943 /dnas_title="GAA-pGEn1-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 3166..5943 /dnas_title="GAA" /gene="GAA" /gene_synonym="LYAG" /db_xref="GeneID:2548" /db_xref="HGNC:4065" /db_xref="MIM:606800" CDS 3166..5943 /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 3267..3267 /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 3166..5943 /note="GAA coding region" misc_difference 5655..5655 /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(5952..7354,1..3132) /source="pGEn1-DEST" /type="Destination Vector" clone 3133..5951 /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 GCCGCCATGA GACTGCTGAC AGCCCTGTTC 3001 GCCTACTTCA TCGTGGCCCT GATCCTGGCC TTCAGCGTGT CCGCCAAGAG CATGCACCAC 3061 CACCATCACC ATCATCACAG CGCCTGGTCC CACCCCCAGT TCGAGAAAGA ATTATCAACA 3121 AGTTTGTACA AAAAAGCAGG CTctgaaaac ttgtactttc aaggcgggca catcctactc 3181 catgatttcc tgctggttcc ccgagagctg agtggctcct ccccagtcct ggaggagact 3241 cacccagctc accagcaggg agccagtaga ccagggcccc gggatgccca ggcacacccc 3301 ggccgtccca gagcagtgcc cacacagtgc gacgtccccc ccaacagccg cttcgattgc 3361 gcccctgaca aggccatcac ccaggaacag tgcgaggccc gcggctgttg ctacatccct 3421 gcaaagcagg ggctgcaggg agcccagatg gggcagccct ggtgcttctt cccacccagc 3481 taccccagct acaagctgga gaacctgagc tcctctgaaa tgggctacac ggccaccctg 3541 acccgtacca cccccacctt cttccccaag gacatcctga ccctgcggct ggacgtgatg 3601 atggagactg agaaccgcct ccacttcacg atcaaagatc cagctaacag gcgctacgag 3661 gtgcccttgg agaccccgca tgtccacagc cgggcaccgt ccccactcta cagcgtggag 3721 ttctccgagg agcccttcgg ggtgatcgtg cgccggcagc tggacggccg cgtgctgctg 3781 aacacgacgg tggcgcccct gttctttgcg gaccagttcc ttcagctgtc cacctcgctg 3841 ccctcgcagt atatcacagg cctcgccgag cacctcagtc ccctgatgct cagcaccagc 3901 tggaccagga tcaccctgtg gaaccgggac cttgcgccca cgcccggtgc gaacctctac 3961 gggtctcacc ctttctacct ggcgctggag gacggcgggt cggcacacgg ggtgttcctg 4021 ctaaacagca atgccatgga tgtggtcctg cagccgagcc ctgcccttag ctggaggtcg 4081 acaggtggga tcctggatgt ctacatcttc ctgggcccag agcccaagag cgtggtgcag 4141 cagtacctgg acgttgtggg atacccgttc atgccgccat actggggcct gggcttccac 4201 ctgtgccgct ggggctactc ctccaccgct atcacccgcc aggtggtgga gaacatgacc 4261 agggcccact tccccctgga cgtccagtgg aacgacctgg actacatgga ctcccggagg 4321 gacttcacgt tcaacaagga tggcttccgg gacttcccgg ccatggtgca ggagctgcac 4381 cagggcggcc ggcgctacat gatgatcgtg gatcctgcca tcagcagctc gggccctgcc 4441 gggagctaca ggccctacga cgagggtctg cggagggggg ttttcatcac caacgagacc 4501 ggccagccgc tgattgggaa ggtatggccc gggtccactg ccttccccga cttcaccaac 4561 cccacagccc tggcctggtg ggaggacatg gtggctgagt tccatgacca ggtgcccttc 4621 gacggcatgt ggattgacat gaacgagcct tccaacttca tcaggggctc tgaggacggc 4681 tgccccaaca atgagctgga gaacccaccc tacgtgcctg gggtggttgg ggggaccctc 4741 caggcggcca ccatctgtgc ctccagccac cagtttctct ccacacacta caacctgcac 4801 aacctctacg gcctgaccga agccatcgcc tcccacaggg cgctggtgaa ggctcggggg 4861 acacgcccat ttgtgatctc ccgctcgacc tttgctggcc acggccgata cgccggccac 4921 tggacggggg acgtgtggag ctcctgggag cagctcgcct cctccgtgcc agaaatcctg 4981 cagtttaacc tgctgggggt gcctctggtc ggggccgacg tctgcggctt cctgggcaac 5041 acctcagagg agctgtgtgt gcgctggacc cagctggggg ccttctaccc cttcatgcgg 5101 aaccacaaca gcctgctcag tctgccccag gagccgtaca gcttcagcga gccggcccag 5161 caggccatga ggaaggccct caccctgcgc tacgcactcc tcccccacct ctacacactg 5221 ttccaccagg cccacgtcgc gggggagacc gtggcccggc ccctcttcct ggagttcccc 5281 aaggactcta gcacctggac tgtggaccac cagctcctgt ggggggaggc cctgctcatc 5341 accccagtgc tccaggccgg gaaggccgaa gtgactggct acttcccctt gggcacatgg 5401 tacgacctgc agacggtgcc agtagaggcc cttggcagcc tcccaccccc acctgcagct 5461 ccccgtgagc cagccatcca cagcgagggg cagtgggtga cgctgccggc ccccctggac 5521 accatcaacg tccacctccg ggctgggtac atcatccccc tgcagggccc tggcctcaca 5581 accacagagt cccgccagca gcccatggcc ctggctgtgg ccctgaccaa gggtggggag 5641 gcccgagggg agcttttctg ggacgatgga gagagcctgg aagtgctgga gcgaggggcc 5701 tacacacagg tcatcttcct ggccaggaat aacacgatcg tgaatgagct ggtacgtgtg 5761 accagtgagg gagctggcct gcagctgcag aaggtgactg tcctgggcgt ggccacggcg 5821 ccccagcagg tcctctccaa cggtgtccct gtctccaact tcacctacag ccccgacacc 5881 aaggtcctgg acatctgtgt ctcgctgttg atgggagagc agtttctcgt cagctggtgt 5941 tagCCAGCTT TCTTGTACAA AGTGGTTGAT AGCTTGGATC CAATCAACCT CTGGATTACA 6001 AAATTTGTGA AAGATTGACT GGTATTCTTA ACTATGTTGC TCCTTTTACG CTATGTGGAT 6061 ACGCTGCTTT AATGCCTTTG TATCATGCTA TTGCTTCCCG TATGGCTTTC ATTTTCTCCT 6121 CCTTGTATAA ATCCTGGTTG CTGTCTCTTT ATGAGGAGTT GTGGCCCGTT GTCAGGCAAC 6181 GTGGCGTGGT GTGCACTGTG TTTGCTGACG CAACCCCCAC TGGTTGGGGC ATTGCCACCA 6241 CCTGTCAGCT CCTTTCCGGG ACTTTCGCTT TCCCCCTCCC TATTGCCACG GCGGAACTCA 6301 TCGCCGCCTG CCTTGCCCGC TGCTGGACAG GGGCTCGGCT GTTGGGCACT GACAATTCCG 6361 TGGTGTTGTC GGGGAAGCTG ACGTCCTTTC CATGGCTGCT CGCCTGTGTT GCCACCTGGA 6421 TTCTGCGCGG GACGTCCTTC TGCTACGTCC CTTCGGCCCT CAATCCAGCG GACCTTCCTT 6481 CCCGCGGCCT GCTGCCGGCT CTGCGGCCTC TTCCGCGTCT TCGAGATCTG CCTCGACTGT 6541 GCCTTCTAGT TGCCAGCCAT CTGTTGTTTG CCCCTCCCCC GTGCCTTCCT TGACCCTGGA 6601 AGGTGCCACT CCCACTGTCC TTTCCTAATA AAATGAGGAA ATTGCATCGC ATTGTCTGAG 6661 TAGGTGTCAT TCTATTCTGG GGGGTGGGGT GGGGCAGGAC AGCAAGGGGG AGGATTGGGA 6721 AGACAATAGC AGGCATGCTG GGGACTCGAG TTAAGGGCGA ATTGCCGATT AGGATCTTCC 6781 TAGAGCATGG CTACGTAGAT AAGTAGCATG GCGGGTTAAT CATTAACTAC AAGGAACCCC 6841 TAGTGATGGA GTTGGCCACT CCCTCTCTGC GCGCTCGCTC GCTCACTGAG GCCGGGCGAC 6901 CAAAGGTCGC CCGACGCCCG GGCTTTGCCC GGGCGGCCTC AGTGAGCGAG CGAGCGCGCA 6961 GCCTTAATTA AGAGCTCATG GCGCGCCTAG GCCTTGACGG CCTTCCGCCA ATTCGCCCTA 7021 TAGTGAGTCG TATTACGTCG CGCTCACTGG CCGTCGTTTT ACAACGTCGT GACTGGGAAA 7081 ACCCTGGCGT TACCCAACTT AATCGCCTTG CAGCACATCC CCCTTTCGCC AGCTGGCGTA 7141 ATAGCGAAGA GGCCCGCACC GAAACGCCCT TCCCAACAGT TGCGCAGCCT GAATGGCGAA 7201 TGGGAGCGCC CTGTAGCGGC CACTCAACCC TATCTCGGTC TATTCTTTTG ATTTATAAGG 7261 GATTTTGCCG ATTTCGGCCT ATTGGTTAAA AAATGAGCTG ATTTAACAAA AATTTAACGC 7321 GAATTTTAAC AAAATATTAA CGCTTACAAT TTAG //