LOCUS GAA-pGEn2-DEST 8079 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(3839..3867) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 6707..7248 /note="WPRE" misc_recomb 6669..6691 /label=attB2 /note="attB2 site" misc_feature complement(7255..7469) /note="BGH pA" misc_feature complement(7557..7686) /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..6668 /dnas_title="GAA-pGEn2-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 3891..6668 /dnas_title="GAA" /gene="GAA" /gene_synonym="LYAG" /db_xref="GeneID:2548" /db_xref="HGNC:4065" /db_xref="MIM:606800" CDS 3891..6668 /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 3992..3992 /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 3891..6668 /note="GAA coding region" misc_difference 6380..6380 /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(6677..8079,1..3857) /source="pGEn2-DEST" /type="Destination Vector" clone 3858..6676 /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 GAATTCgcAT 3841 CAACAAGTTT GTACAAAAAA GCAGGCTctg aaaacttgta ctttcaaggc gggcacatcc 3901 tactccatga tttcctgctg gttccccgag agctgagtgg ctcctcccca gtcctggagg 3961 agactcaccc agctcaccag cagggagcca gtagaccagg gccccgggat gcccaggcac 4021 accccggccg tcccagagca gtgcccacac agtgcgacgt cccccccaac agccgcttcg 4081 attgcgcccc tgacaaggcc atcacccagg aacagtgcga ggcccgcggc tgttgctaca 4141 tccctgcaaa gcaggggctg cagggagccc agatggggca gccctggtgc ttcttcccac 4201 ccagctaccc cagctacaag ctggagaacc tgagctcctc tgaaatgggc tacacggcca 4261 ccctgacccg taccaccccc accttcttcc ccaaggacat cctgaccctg cggctggacg 4321 tgatgatgga gactgagaac cgcctccact tcacgatcaa agatccagct aacaggcgct 4381 acgaggtgcc cttggagacc ccgcatgtcc acagccgggc accgtcccca ctctacagcg 4441 tggagttctc cgaggagccc ttcggggtga tcgtgcgccg gcagctggac ggccgcgtgc 4501 tgctgaacac gacggtggcg cccctgttct ttgcggacca gttccttcag ctgtccacct 4561 cgctgccctc gcagtatatc acaggcctcg ccgagcacct cagtcccctg atgctcagca 4621 ccagctggac caggatcacc ctgtggaacc gggaccttgc gcccacgccc ggtgcgaacc 4681 tctacgggtc tcaccctttc tacctggcgc tggaggacgg cgggtcggca cacggggtgt 4741 tcctgctaaa cagcaatgcc atggatgtgg tcctgcagcc gagccctgcc cttagctgga 4801 ggtcgacagg tgggatcctg gatgtctaca tcttcctggg cccagagccc aagagcgtgg 4861 tgcagcagta cctggacgtt gtgggatacc cgttcatgcc gccatactgg ggcctgggct 4921 tccacctgtg ccgctggggc tactcctcca ccgctatcac ccgccaggtg gtggagaaca 4981 tgaccagggc ccacttcccc ctggacgtcc agtggaacga cctggactac atggactccc 5041 ggagggactt cacgttcaac aaggatggct tccgggactt cccggccatg gtgcaggagc 5101 tgcaccaggg cggccggcgc tacatgatga tcgtggatcc tgccatcagc agctcgggcc 5161 ctgccgggag ctacaggccc tacgacgagg gtctgcggag gggggttttc atcaccaacg 5221 agaccggcca gccgctgatt gggaaggtat ggcccgggtc cactgccttc cccgacttca 5281 ccaaccccac agccctggcc tggtgggagg acatggtggc tgagttccat gaccaggtgc 5341 ccttcgacgg catgtggatt gacatgaacg agccttccaa cttcatcagg ggctctgagg 5401 acggctgccc caacaatgag ctggagaacc caccctacgt gcctggggtg gttgggggga 5461 ccctccaggc ggccaccatc tgtgcctcca gccaccagtt tctctccaca cactacaacc 5521 tgcacaacct ctacggcctg accgaagcca tcgcctccca cagggcgctg gtgaaggctc 5581 gggggacacg cccatttgtg atctcccgct cgacctttgc tggccacggc cgatacgccg 5641 gccactggac gggggacgtg tggagctcct gggagcagct cgcctcctcc gtgccagaaa 5701 tcctgcagtt taacctgctg ggggtgcctc tggtcggggc cgacgtctgc ggcttcctgg 5761 gcaacacctc agaggagctg tgtgtgcgct ggacccagct gggggccttc taccccttca 5821 tgcggaacca caacagcctg ctcagtctgc cccaggagcc gtacagcttc agcgagccgg 5881 cccagcaggc catgaggaag gccctcaccc tgcgctacgc actcctcccc cacctctaca 5941 cactgttcca ccaggcccac gtcgcggggg agaccgtggc ccggcccctc ttcctggagt 6001 tccccaagga ctctagcacc tggactgtgg accaccagct cctgtggggg gaggccctgc 6061 tcatcacccc agtgctccag gccgggaagg ccgaagtgac tggctacttc cccttgggca 6121 catggtacga cctgcagacg gtgccagtag aggcccttgg cagcctccca cccccacctg 6181 cagctccccg tgagccagcc atccacagcg aggggcagtg ggtgacgctg ccggcccccc 6241 tggacaccat caacgtccac ctccgggctg ggtacatcat ccccctgcag ggccctggcc 6301 tcacaaccac agagtcccgc cagcagccca tggccctggc tgtggccctg accaagggtg 6361 gggaggcccg aggggagctt ttctgggacg atggagagag cctggaagtg ctggagcgag 6421 gggcctacac acaggtcatc ttcctggcca ggaataacac gatcgtgaat gagctggtac 6481 gtgtgaccag tgagggagct ggcctgcagc tgcagaaggt gactgtcctg ggcgtggcca 6541 cggcgcccca gcaggtcctc tccaacggtg tccctgtctc caacttcacc tacagccccg 6601 acaccaaggt cctggacatc tgtgtctcgc tgttgatggg agagcagttt ctcgtcagct 6661 ggtgttagCC AGCTTTCTTG TACAAAGTGG TTGATAGCTT GGATCCAATC AACCTCTGGA 6721 TTACAAAATT TGTGAAAGAT TGACTGGTAT TCTTAACTAT GTTGCTCCTT TTACGCTATG 6781 TGGATACGCT GCTTTAATGC CTTTGTATCA TGCTATTGCT TCCCGTATGG CTTTCATTTT 6841 CTCCTCCTTG TATAAATCCT GGTTGCTGTC TCTTTATGAG GAGTTGTGGC CCGTTGTCAG 6901 GCAACGTGGC GTGGTGTGCA CTGTGTTTGC TGACGCAACC CCCACTGGTT GGGGCATTGC 6961 CACCACCTGT CAGCTCCTTT CCGGGACTTT CGCTTTCCCC CTCCCTATTG CCACGGCGGA 7021 ACTCATCGCC GCCTGCCTTG CCCGCTGCTG GACAGGGGCT CGGCTGTTGG GCACTGACAA 7081 TTCCGTGGTG TTGTCGGGGA AGCTGACGTC CTTTCCATGG CTGCTCGCCT GTGTTGCCAC 7141 CTGGATTCTG CGCGGGACGT CCTTCTGCTA CGTCCCTTCG GCCCTCAATC CAGCGGACCT 7201 TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG GCCTCTTCCG CGTCTTCGAG ATCTGCCTCG 7261 ACTGTGCCTT CTAGTTGCCA GCCATCTGTT GTTTGCCCCT CCCCCGTGCC TTCCTTGACC 7321 CTGGAAGGTG CCACTCCCAC TGTCCTTTCC TAATAAAATG AGGAAATTGC ATCGCATTGT 7381 CTGAGTAGGT GTCATTCTAT TCTGGGGGGT GGGGTGGGGC AGGACAGCAA GGGGGAGGAT 7441 TGGGAAGACA ATAGCAGGCA TGCTGGGGAC TCGAGTTAAG GGCGAATTGC CGATTAGGAT 7501 CTTCCTAGAG CATGGCTACG TAGATAAGTA GCATGGCGGG TTAATCATTA ACTACAAGGA 7561 ACCCCTAGTG ATGGAGTTGG CCACTCCCTC TCTGCGCGCT CGCTCGCTCA CTGAGGCCGG 7621 GCGACCAAAG GTCGCCCGAC GCCCGGGCTT TGCCCGGGCG GCCTCAGTGA GCGAGCGAGC 7681 GCGCAGCCTT AATTAAGAGC TCATGGCGCG CCTAGGCCTT GACGGCCTTC CGCCAATTCG 7741 CCCTATAGTG AGTCGTATTA CGTCGCGCTC ACTGGCCGTC GTTTTACAAC GTCGTGACTG 7801 GGAAAACCCT GGCGTTACCC AACTTAATCG CCTTGCAGCA CATCCCCCTT TCGCCAGCTG 7861 GCGTAATAGC GAAGAGGCCC GCACCGAAAC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG 7921 GCGAATGGGA GCGCCCTGTA GCGGCCACTC AACCCTATCT CGGTCTATTC TTTTGATTTA 7981 TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG AGCTGATTTA ACAAAAATTT 8041 AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTAG //