LOCUS MANBA-pGEn3-DEST 8592 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 7220..7761 /note="WPRE" misc_recomb 7182..7204 /label=attB2 /note="attB2 site" misc_feature complement(7768..7982) /note="BGH pA" misc_feature complement(8070..8199) /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..7181 /dnas_title="MANBA-pGEn3-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:23037 IMAGE:4905535" /tissue_type="Muscle, rhabdomyosarcoma" /clone_lib="NIH_MGC_17" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 4593..7181 /dnas_title="MANBA" /gene="MANBA" /gene_synonym="MANB1" /db_xref="GeneID:4126" /db_xref="HGNC:6831" /db_xref="MIM:609489" CDS 4593..7181 /dnas_title="mannosidase, beta A, lysosomal" /gene="MANBA" /gene_synonym="MANB1" /codon_start=1 /product="mannosidase, beta A, lysosomal" /protein_id="AAH15743.1" /db_xref="GI:16041731" /db_xref="GeneID:4126" /db_xref="HGNC:6831" /db_xref="MIM:609489" /translation="MRLHLLLLLALCGAGTTAAELSYSLRGNWSICNGNGSLELPGAV PGCVHSALFQQGLIQDSYYRFNDLNYRWVSLDNWTYSKEFKIPFEISKWQKVNLILEG VDTVSKILFNEVTIGETDNMFNRYSFDITNVVRDVNSIELRFQSAVLYAAQQSKAHTR YQVPPDCPPLVQKGECHVNFVRKEQCSFSWDWGPSFPTQGIWKDVRIEAYNICHLNYF TFSPIYDKSAQEWNLEIESTFDVVSSKPVGGQVIVAIPKLQTQQTYSIELQPGKRIVE LFVNISKNITVETWWPHGHGNQTGYNMTVLFELDGGLNIEKSAKVYFRTVELIEEPIK GSPGLSFYFKINGFPIFLKGSNWIPADSFQDRVTSELLRLLLQSVVDANMNTLRVWGG GIYEQDEFYELCDELGIMVWQDFMFACALYPTDQGFLDSVTAEVAYQIKRLKSHPSII IWSGNNENEEALMMNWYHISFTDRPIYIKDYVTLYVKNIRELVLAGDKSRPFITSSPT NGAETVAEAWVSQNPNSNYFGDVHFYDYISDCWNWKVFPKARFASEYGYQSWPSFSTL EKVSSTEDWSFNSKFSLHRQHHEGGNKQMLYQAGLHFKLPQSTDPLRTFKDTIYLTQV MQAQCVKTETEFYRRSRSEIVDQQGHTMGALYWQLNDIWQAPSWASLEYGGKWKMLHY FAQNFFAPLLPVGFENENMFYIYGVSDLHSDYSMTLSVRVHTWSSLEPVCSRVTERFV MKGGEAVCLYEEPVSELLRRCGNCTRESCVVSFYLSADHELLSPTNYHFLSSPKEAVG LCKAQITAIISQQGDIFVFDLETSAVAPFVWLDVGSIPGRFSDNGFLMTEKTRTILFY PWEPTSKNELEQSFHVTSLTDIY" misc_feature 4593..7181 /note="MANB1" misc_difference 6643..6643 /dnas_title="MANBA" /gene="MANBA" /gene_synonym="MANB1" /note="'T' in cDNA is 'C' in the human genome; amino acid difference: 'M' in cDNA, 'T' in the human genome. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." misc_difference 6909..6909 /dnas_title="MANBA" /gene="MANBA" /gene_synonym="MANB1" /note="'C' in cDNA is 'T' in the human genome; no amino acid change." vector join(7190..8592,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..7189 /source="MANBA-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 gcgccgcgga gctcagttac agcttgcgtg 4621 gcaactggag catctgcaat gggaacggct cgctggagct gcccggggcg gtccctggct 4681 gcgtgcacag cgccttgttc cagcagggcc tgatccagga ttcttactac agatttaatg 4741 accttaacta cagatgggtc tctttggata actggaccta tagcaaagaa tttaaaatcc 4801 cctttgaaat tagcaaatgg caaaaagtaa atttgattct tgagggagtg gatacggttt 4861 caaaaatcct gttcaatgaa gtcactattg gggaaacaga caatatgttc aatagatata 4921 gctttgatat taccaacgtg gtcagggacg tgaactccat tgagctgcgt ttccagtcag 4981 cggtgttgta tgcagcacag cagagcaaag ctcacactcg ctaccaggtt cccccagact 5041 gccctccact tgtgcagaag ggtgaatgcc atgtcaactt tgttcggaag gagcaatgtt 5101 cctttagttg ggactggggg ccttcctttc ctacccaggg aatctggaaa gatgttagaa 5161 ttgaagccta taatatttgt cacctgaact acttcacatt ttccccaata tatgataaga 5221 gtgcccagga gtggaatctg gaaatagagt ctacatttga tgttgtcagc tcaaagccag 5281 ttggtggtca agtgatcgta gccatcccta agttgcaaac acaacagaca tacagcattg 5341 aacttcaacc tgggaaaagg attgttgagc tatttgtgaa cattagcaag aatattactg 5401 tagaaacttg gtggcctcat ggacatggaa accagactgg gtacaacatg actgttcttt 5461 ttgaactgga tggaggctta aatattgaaa aatcagctaa ggtttatttt aggacagtgg 5521 aacttataga agagcctata aaagggtctc ctggtttgag tttctatttc aaaattaatg 5581 gatttcccat atttctaaaa ggctcaaact ggatcccagc agattcattc caggaccgag 5641 taacctctga gttgttacgg ctccttttac agtctgttgt ggatgctaat atgaatactc 5701 ttcgggtttg gggaggagga atttatgagc aggatgaatt ctatgaactc tgtgatgaac 5761 taggaataat ggtatggcag gattttatgt ttgcctgtgc cctttatcca actgatcagg 5821 gcttcctgga ttcagtgaca gcagaagttg cctaccagat caagagactg aaatctcatc 5881 cttctatcat catatggagt ggcaataatg aaaatgagga ggcgctgatg atgaattggt 5941 atcatatcag tttcactgac cggccaatct acatcaagga ctatgtgaca ctctatgtga 6001 aaaacatcag agagctcgta ctggcaggag acaagagtcg tccttttatt acgtccagtc 6061 ctacaaatgg ggctgaaact gttgcagaag cctgggtctc tcaaaaccct aatagcaatt 6121 attttggtga tgtacatttt tatgactata tcagtgattg ctggaactgg aaagttttcc 6181 caaaagctcg atttgcatct gaatatggat atcagtcctg gccgtccttc agtacattag 6241 aaaaggtctc gtctacagag gactggtctt tcaatagcaa gttttcactt catcgacaac 6301 atcacgaagg tggtaacaaa caaatgcttt atcaggctgg acttcatttc aaactccccc 6361 aaagcacaga tccattacgc acatttaaag ataccatcta ccttactcag gtgatgcagg 6421 cccagtgtgt caaaacagaa actgaattct accgccgtag tcgcagcgag atagtggatc 6481 agcaagggca cacgatgggg gcactttatt ggcagttgaa tgacatctgg caagctcctt 6541 cctgggcttc tcttgagtac ggaggaaagt ggaaaatgct tcattacttt gctcagaatt 6601 tctttgctcc actgttgcca gtaggctttg agaatgaaaa catgttctat atctatggtg 6661 tgtcagatct tcactcggat tattcgatga cactcagtgt gagagtccat acatggagct 6721 ccctggagcc cgtgtgctct cgtgtgactg aacgttttgt gatgaaagga ggagaggctg 6781 tctgccttta tgaggagcca gtgtctgaat tgctgaggag atgtgggaat tgcacacggg 6841 aaagctgtgt ggtttccttt tacctttcag ctgaccatga actcctgagc ccgaccaact 6901 accacttcct gtcctcaccg aaggaggccg tggggctctg caaggcgcag atcactgcca 6961 tcatctctca gcaaggtgac atatttgttt ttgacctgga gacctcagct gtcgctccct 7021 ttgtttggtt ggatgtagga agcatcccag ggagatttag tgacaatggt ttcctcatga 7081 ctgagaagac acgaactata ttattttacc cttgggagcc caccagcaag aatgagttgg 7141 agcaatcttt tcatgtgacc tccttaacag atatttactg aCCAGCTTTC TTGTACAAAG 7201 TGGTTGATAG CTTGGATCCA ATCAACCTCT GGATTACAAA ATTTGTGAAA GATTGACTGG 7261 TATTCTTAAC TATGTTGCTC CTTTTACGCT ATGTGGATAC GCTGCTTTAA TGCCTTTGTA 7321 TCATGCTATT GCTTCCCGTA TGGCTTTCAT TTTCTCCTCC TTGTATAAAT CCTGGTTGCT 7381 GTCTCTTTAT GAGGAGTTGT GGCCCGTTGT CAGGCAACGT GGCGTGGTGT GCACTGTGTT 7441 TGCTGACGCA ACCCCCACTG GTTGGGGCAT TGCCACCACC TGTCAGCTCC TTTCCGGGAC 7501 TTTCGCTTTC CCCCTCCCTA TTGCCACGGC GGAACTCATC GCCGCCTGCC TTGCCCGCTG 7561 CTGGACAGGG GCTCGGCTGT TGGGCACTGA CAATTCCGTG GTGTTGTCGG GGAAGCTGAC 7621 GTCCTTTCCA TGGCTGCTCG CCTGTGTTGC CACCTGGATT CTGCGCGGGA CGTCCTTCTG 7681 CTACGTCCCT TCGGCCCTCA ATCCAGCGGA CCTTCCTTCC CGCGGCCTGC TGCCGGCTCT 7741 GCGGCCTCTT CCGCGTCTTC GAGATCTGCC TCGACTGTGC CTTCTAGTTG CCAGCCATCT 7801 GTTGTTTGCC CCTCCCCCGT GCCTTCCTTG ACCCTGGAAG GTGCCACTCC CACTGTCCTT 7861 TCCTAATAAA ATGAGGAAAT TGCATCGCAT TGTCTGAGTA GGTGTCATTC TATTCTGGGG 7921 GGTGGGGTGG GGCAGGACAG CAAGGGGGAG GATTGGGAAG ACAATAGCAG GCATGCTGGG 7981 GACTCGAGTT AAGGGCGAAT TGCCGATTAG GATCTTCCTA GAGCATGGCT ACGTAGATAA 8041 GTAGCATGGC GGGTTAATCA TTAACTACAA GGAACCCCTA GTGATGGAGT TGGCCACTCC 8101 CTCTCTGCGC GCTCGCTCGC TCACTGAGGC CGGGCGACCA AAGGTCGCCC GACGCCCGGG 8161 CTTTGCCCGG GCGGCCTCAG TGAGCGAGCG AGCGCGCAGC CTTAATTAAG AGCTCATGGC 8221 GCGCCTAGGC CTTGACGGCC TTCCGCCAAT TCGCCCTATA GTGAGTCGTA TTACGTCGCG 8281 CTCACTGGCC GTCGTTTTAC AACGTCGTGA CTGGGAAAAC CCTGGCGTTA CCCAACTTAA 8341 TCGCCTTGCA GCACATCCCC CTTTCGCCAG CTGGCGTAAT AGCGAAGAGG CCCGCACCGA 8401 AACGCCCTTC CCAACAGTTG CGCAGCCTGA ATGGCGAATG GGAGCGCCCT GTAGCGGCCA 8461 CTCAACCCTA TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT 8521 TGGTTAAAAA ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG 8581 CTTACAATTT AG //