LOCUS MANBA-pGEn1-DEST 7165 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 5793..6334 /note="WPRE" misc_recomb 5755..5777 /label=attB2 /note="attB2 site" misc_feature complement(6341..6555) /note="BGH pA" misc_feature complement(6643..6772) /note="3'-ITR" misc_feature 3143..3165 /note="TEV site" source 3166..5754 /dnas_title="MANBA-pGEn1-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 3166..5754 /dnas_title="MANBA" /gene="MANBA" /gene_synonym="MANB1" /db_xref="GeneID:4126" /db_xref="HGNC:6831" /db_xref="MIM:609489" CDS 3166..5754 /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 3166..5754 /note="MANB1" misc_difference 5216..5216 /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 5482..5482 /dnas_title="MANBA" /gene="MANBA" /gene_synonym="MANB1" /note="'C' in cDNA is 'T' in the human genome; no amino acid change." vector join(5763..7165,1..3132) /source="pGEn1-DEST" /type="Destination Vector" clone 3133..5762 /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 GCCGCCATGA GACTGCTGAC AGCCCTGTTC 3001 GCCTACTTCA TCGTGGCCCT GATCCTGGCC TTCAGCGTGT CCGCCAAGAG CATGCACCAC 3061 CACCATCACC ATCATCACAG CGCCTGGTCC CACCCCCAGT TCGAGAAAGA ATTATCAACA 3121 AGTTTGTACA AAAAAGCAGG CTctgaaaac ttgtactttc aaggcgccgc ggagctcagt 3181 tacagcttgc gtggcaactg gagcatctgc aatgggaacg gctcgctgga gctgcccggg 3241 gcggtccctg gctgcgtgca cagcgccttg ttccagcagg gcctgatcca ggattcttac 3301 tacagattta atgaccttaa ctacagatgg gtctctttgg ataactggac ctatagcaaa 3361 gaatttaaaa tcccctttga aattagcaaa tggcaaaaag taaatttgat tcttgaggga 3421 gtggatacgg tttcaaaaat cctgttcaat gaagtcacta ttggggaaac agacaatatg 3481 ttcaatagat atagctttga tattaccaac gtggtcaggg acgtgaactc cattgagctg 3541 cgtttccagt cagcggtgtt gtatgcagca cagcagagca aagctcacac tcgctaccag 3601 gttcccccag actgccctcc acttgtgcag aagggtgaat gccatgtcaa ctttgttcgg 3661 aaggagcaat gttcctttag ttgggactgg gggccttcct ttcctaccca gggaatctgg 3721 aaagatgtta gaattgaagc ctataatatt tgtcacctga actacttcac attttcccca 3781 atatatgata agagtgccca ggagtggaat ctggaaatag agtctacatt tgatgttgtc 3841 agctcaaagc cagttggtgg tcaagtgatc gtagccatcc ctaagttgca aacacaacag 3901 acatacagca ttgaacttca acctgggaaa aggattgttg agctatttgt gaacattagc 3961 aagaatatta ctgtagaaac ttggtggcct catggacatg gaaaccagac tgggtacaac 4021 atgactgttc tttttgaact ggatggaggc ttaaatattg aaaaatcagc taaggtttat 4081 tttaggacag tggaacttat agaagagcct ataaaagggt ctcctggttt gagtttctat 4141 ttcaaaatta atggatttcc catatttcta aaaggctcaa actggatccc agcagattca 4201 ttccaggacc gagtaacctc tgagttgtta cggctccttt tacagtctgt tgtggatgct 4261 aatatgaata ctcttcgggt ttggggagga ggaatttatg agcaggatga attctatgaa 4321 ctctgtgatg aactaggaat aatggtatgg caggatttta tgtttgcctg tgccctttat 4381 ccaactgatc agggcttcct ggattcagtg acagcagaag ttgcctacca gatcaagaga 4441 ctgaaatctc atccttctat catcatatgg agtggcaata atgaaaatga ggaggcgctg 4501 atgatgaatt ggtatcatat cagtttcact gaccggccaa tctacatcaa ggactatgtg 4561 acactctatg tgaaaaacat cagagagctc gtactggcag gagacaagag tcgtcctttt 4621 attacgtcca gtcctacaaa tggggctgaa actgttgcag aagcctgggt ctctcaaaac 4681 cctaatagca attattttgg tgatgtacat ttttatgact atatcagtga ttgctggaac 4741 tggaaagttt tcccaaaagc tcgatttgca tctgaatatg gatatcagtc ctggccgtcc 4801 ttcagtacat tagaaaaggt ctcgtctaca gaggactggt ctttcaatag caagttttca 4861 cttcatcgac aacatcacga aggtggtaac aaacaaatgc tttatcaggc tggacttcat 4921 ttcaaactcc cccaaagcac agatccatta cgcacattta aagataccat ctaccttact 4981 caggtgatgc aggcccagtg tgtcaaaaca gaaactgaat tctaccgccg tagtcgcagc 5041 gagatagtgg atcagcaagg gcacacgatg ggggcacttt attggcagtt gaatgacatc 5101 tggcaagctc cttcctgggc ttctcttgag tacggaggaa agtggaaaat gcttcattac 5161 tttgctcaga atttctttgc tccactgttg ccagtaggct ttgagaatga aaacatgttc 5221 tatatctatg gtgtgtcaga tcttcactcg gattattcga tgacactcag tgtgagagtc 5281 catacatgga gctccctgga gcccgtgtgc tctcgtgtga ctgaacgttt tgtgatgaaa 5341 ggaggagagg ctgtctgcct ttatgaggag ccagtgtctg aattgctgag gagatgtggg 5401 aattgcacac gggaaagctg tgtggtttcc ttttaccttt cagctgacca tgaactcctg 5461 agcccgacca actaccactt cctgtcctca ccgaaggagg ccgtggggct ctgcaaggcg 5521 cagatcactg ccatcatctc tcagcaaggt gacatatttg tttttgacct ggagacctca 5581 gctgtcgctc cctttgtttg gttggatgta ggaagcatcc cagggagatt tagtgacaat 5641 ggtttcctca tgactgagaa gacacgaact atattatttt acccttggga gcccaccagc 5701 aagaatgagt tggagcaatc ttttcatgtg acctccttaa cagatattta ctgaCCAGCT 5761 TTCTTGTACA AAGTGGTTGA TAGCTTGGAT CCAATCAACC TCTGGATTAC AAAATTTGTG 5821 AAAGATTGAC TGGTATTCTT AACTATGTTG CTCCTTTTAC GCTATGTGGA TACGCTGCTT 5881 TAATGCCTTT GTATCATGCT ATTGCTTCCC GTATGGCTTT CATTTTCTCC TCCTTGTATA 5941 AATCCTGGTT GCTGTCTCTT TATGAGGAGT TGTGGCCCGT TGTCAGGCAA CGTGGCGTGG 6001 TGTGCACTGT GTTTGCTGAC GCAACCCCCA CTGGTTGGGG CATTGCCACC ACCTGTCAGC 6061 TCCTTTCCGG GACTTTCGCT TTCCCCCTCC CTATTGCCAC GGCGGAACTC ATCGCCGCCT 6121 GCCTTGCCCG CTGCTGGACA GGGGCTCGGC TGTTGGGCAC TGACAATTCC GTGGTGTTGT 6181 CGGGGAAGCT GACGTCCTTT CCATGGCTGC TCGCCTGTGT TGCCACCTGG ATTCTGCGCG 6241 GGACGTCCTT CTGCTACGTC CCTTCGGCCC TCAATCCAGC GGACCTTCCT TCCCGCGGCC 6301 TGCTGCCGGC TCTGCGGCCT CTTCCGCGTC TTCGAGATCT GCCTCGACTG TGCCTTCTAG 6361 TTGCCAGCCA TCTGTTGTTT GCCCCTCCCC CGTGCCTTCC TTGACCCTGG AAGGTGCCAC 6421 TCCCACTGTC CTTTCCTAAT AAAATGAGGA AATTGCATCG CATTGTCTGA GTAGGTGTCA 6481 TTCTATTCTG GGGGGTGGGG TGGGGCAGGA CAGCAAGGGG GAGGATTGGG AAGACAATAG 6541 CAGGCATGCT GGGGACTCGA GTTAAGGGCG AATTGCCGAT TAGGATCTTC CTAGAGCATG 6601 GCTACGTAGA TAAGTAGCAT GGCGGGTTAA TCATTAACTA CAAGGAACCC CTAGTGATGG 6661 AGTTGGCCAC TCCCTCTCTG CGCGCTCGCT CGCTCACTGA GGCCGGGCGA CCAAAGGTCG 6721 CCCGACGCCC GGGCTTTGCC CGGGCGGCCT CAGTGAGCGA GCGAGCGCGC AGCCTTAATT 6781 AAGAGCTCAT GGCGCGCCTA GGCCTTGACG GCCTTCCGCC AATTCGCCCT ATAGTGAGTC 6841 GTATTACGTC GCGCTCACTG GCCGTCGTTT TACAACGTCG TGACTGGGAA AACCCTGGCG 6901 TTACCCAACT TAATCGCCTT GCAGCACATC CCCCTTTCGC CAGCTGGCGT AATAGCGAAG 6961 AGGCCCGCAC CGAAACGCCC TTCCCAACAG TTGCGCAGCC TGAATGGCGA ATGGGAGCGC 7021 CCTGTAGCGG CCACTCAACC CTATCTCGGT CTATTCTTTT GATTTATAAG GGATTTTGCC 7081 GATTTCGGCC TATTGGTTAA AAAATGAGCT GATTTAACAA AAATTTAACG CGAATTTTAA 7141 CAAAATATTA ACGCTTACAA TTTAG //