LOCUS GLB1-pGEc1-DEST 6526 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6004..6133) /note="3'-ITR" misc_feature complement(5702..5916) /note="BGH pA" rep_origin 1137..1804 /dnas_title="Col E1 origin" /label=Col E1 origin /vntifkey="33" CDS 129..989 /dnas_title="AmpR" /label=AmpR /vntifkey="4" misc_feature 1998..2127 /note="5'ITR" misc_feature 2208..2790 /note="CMV Promoter" misc_feature 2828..2960 /note="Artificial Intron" misc_feature 5154..5695 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_recomb 5059..5081 /label=attB2 /note="attB2 site" misc_feature 5091..5114 /note="8xHis Tag" misc_feature 5118..5141 /note="StrepTag" PCR_frame 3002..5059 /note="Reading Frame" /set="Set 1" source 3006..5036 /dnas_title="GLB1-pGEc1-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:2315 IMAGE:2988086" /tissue_type="Colon, adenocarcinoma" /clone_lib="NIH_MGC_15" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 3006..5036 /dnas_title="GLB1" /gene="GLB1" /db_xref="GeneID:2720" /db_xref="HGNC:4298" /db_xref="MIM:230500" CDS 3006..5036 /dnas_title="galactosidase, beta 1" /gene="GLB1" /codon_start=1 /product="galactosidase, beta 1" /protein_id="AAH07493.1" /db_xref="GI:13960104" /db_xref="GeneID:2720" /db_xref="HGNC:4298" /db_xref="MIM:230500" /translation="MPGFLVRILLLLLVLLLLGPTRGLRNATQRMFEIDYSRDSFLKD GQPFRYISGSIHYSRVPRFYWKDRLLKMKMAGLNAIQTYVPWNFHEPWPGQYQFSEDH DVEYFLRLAHELGLLVILRPGPYICAEWEMGGLPAWLLEKESILLRSSDPDYLAAVDK WLGVLLPKMKPLLYQNGGPVITVQVENEYGSYFACDFDYLRFLQKRFRHHLGDDVVLF TTDGAHKTFLKCGALQGLYTTVDFGTGSNITDAFLSQRKCEPKGPLINSEFYTGWLDH WGQPHSTIKTEAVASSLYDILARGASVNLYMFIGGTNFAYWNGANSPYAAQPTSYDYD APLSEAGDLTEKYFALRNIIQKFEKVPEGPIPPSTPKFAYGKVTLEKLKTVGAALDIL CPSGPIKSLYPLTFIQVKQHYGFVLYRTTLPQDCSNPAPLSSPLNGVHDRAYVAVDGI PQGVLERNNVITLNITGKAGATLDLLVENMGRVNYGAYINDFKGLVSNLTLSSNILTD WTIFPLDTEDAVRSHLGGWGHRDSGHHDEAWAHNSSNYTLPAFYMGNFSIPSGIPDLP QDTFIQFPGWTKGQVWINGFNLGRYWPARGPQLTLFVPQHILMTSAPNTITVLELEWA PCSSDDPELCAVTFVDRPVIGSSVTYDHPSKPVEKRLMPPPPQKNKDSWLDHV" misc_difference 3034..3034 /dnas_title="GLB1" /gene="GLB1" /note="'T' in cDNA is 'C' in the human genome; amino acid difference: 'L' in cDNA, 'P' in the human genome. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." misc_difference 3039..3039 /dnas_title="GLB1" /gene="GLB1" /note="'C' in cDNA is 'T' in the human genome; no amino acid change." misc_feature 3006..5036 /note="GLB1 coding region" misc_difference 4566..4566 /dnas_title="GLB1" /gene="GLB1" /note="'C' in cDNA is 'T' in the human genome; amino acid difference: 'R' in cDNA, 'C' in the human genome." misc_feature 5037..5057 /note="TEV site" PCR_primer 2992..3001 /dnas_title="GTACAAAAAAGCAGGCTCATGGAACAAGGACAGCTCTTCC" /primer="CAGCTCTTCC" /set="Set 1" /pair="Pair 1" /current=1 /original="GTACAAAAAAGCAGGCTCATGGAACAAGGACAGCTCTTCC" PCR_primer complement(3002..5066) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..5066) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(5067..6526,1..2991) /source="pGEc1-DEST" /type="Destination Vector" clone 2992..5066 /source="GLB1-C-term-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 GCCATCACAA GTTTGTACAA AAAAGCAGGC 3001 TCGCCatgcc ggggttcctg gttcgcatcc tccttctgct gctggttctg ctgcttctgg 3061 gccctacgcg cggcttgcgc aatgccaccc agaggatgtt tgaaattgac tatagccggg 3121 actccttcct caaggatggc cagccatttc gctacatctc aggaagcatt cactactccc 3181 gtgtgccccg cttctactgg aaggaccggc tgctgaagat gaagatggct gggctgaacg 3241 ccatccagac gtatgtgccc tggaactttc atgagccctg gccaggacag taccagtttt 3301 ctgaggacca tgatgtggaa tattttcttc ggctggctca tgagctggga ctgctggtta 3361 tcctgaggcc cgggccctac atctgtgcag agtgggaaat gggaggatta cctgcttggc 3421 tgctagagaa agagtctatt cttctccgct cctccgaccc agattacctg gcagctgtgg 3481 acaagtggtt gggagtcctt ctgcccaaga tgaagcctct cctctatcag aatggagggc 3541 cagttataac agtgcaggtt gaaaatgaat atggcagcta ctttgcctgt gattttgact 3601 acctgcgctt cctgcagaag cgctttcgcc accatctggg ggatgatgtg gttctgttta 3661 ccactgatgg agcacataaa acattcctga aatgtggggc cctgcagggc ctctacacca 3721 cggtggactt tggaacaggc agcaacatca cagatgcttt cctaagccag aggaagtgtg 3781 agcccaaagg acccttgatc aattctgaat tctatactgg ctggctagat cactggggcc 3841 aacctcactc cacaatcaag accgaagcag tggcttcctc cctctatgat atacttgccc 3901 gtggggcgag tgtgaacttg tacatgttta taggtgggac caattttgcc tattggaatg 3961 gggccaactc accctatgca gcacagccca ccagctacga ctatgatgcc ccactgagtg 4021 aggctgggga cctcactgag aagtattttg ctctgcgaaa catcatccag aagtttgaaa 4081 aagtaccaga aggtcctatc cctccatcta caccaaagtt tgcatatgga aaggtcactt 4141 tggaaaagtt aaagacagtg ggagcagctc tggacattct gtgtccctct gggcccatca 4201 aaagccttta tcccttgaca tttatccagg tgaaacagca ttatgggttt gtgctgtacc 4261 ggacaacact tcctcaagat tgcagcaacc cagcacctct ctcttcaccc ctcaatggag 4321 tccacgatcg agcatatgtt gctgtggatg ggatccccca gggagtcctt gagcgaaaca 4381 atgtgatcac tctgaacata acagggaaag ctggagccac tctggacctt ctggtagaga 4441 acatgggacg tgtgaactat ggtgcatata tcaacgattt taagggtttg gtttctaacc 4501 tgactctcag ttccaatatc ctcacggact ggacgatctt tccactggac actgaggatg 4561 cagtgcgcag ccacctgggg ggctggggac accgtgacag tggccaccat gatgaagcct 4621 gggcccacaa ctcatccaac tacacgctcc cggcctttta tatggggaac ttctccattc 4681 ccagtgggat cccagacttg ccccaggaca cctttatcca gtttcctgga tggaccaagg 4741 gccaggtctg gattaatggc tttaaccttg gccgctattg gccagcccgg ggccctcagt 4801 tgaccttgtt tgtgccccag cacatcctga tgacctcggc cccaaacacc atcaccgtgc 4861 tggaactgga gtgggcaccc tgcagcagtg atgatccaga actatgtgct gtgacgttcg 4921 tggacaggcc agttattggc tcatctgtga cctacgatca tccctccaaa cctgttgaaa 4981 aaagactcat gcccccaccc ccgcaaaaaa acaaagattc atggctggac catgtagaaa 5041 acttgtactt tcaaggctCC AGCTTTCTTG TACAAAGTGG TGATGGCCGC CATCATCATC 5101 ATCATCATCA TCATGCGTGG AGCCACCCGC AGTTCGAAAA ATAATAAGGA TCCAATCAAC 5161 CTCTGGATTA CAAAATTTGT GAAAGATTGA CTGGTATTCT TAACTATGTT GCTCCTTTTA 5221 CGCTATGTGG ATACGCTGCT TTAATGCCTT TGTATCATGC TATTGCTTCC CGTATGGCTT 5281 TCATTTTCTC CTCCTTGTAT AAATCCTGGT TGCTGTCTCT TTATGAGGAG TTGTGGCCCG 5341 TTGTCAGGCA ACGTGGCGTG GTGTGCACTG TGTTTGCTGA CGCAACCCCC ACTGGTTGGG 5401 GCATTGCCAC CACCTGTCAG CTCCTTTCCG GGACTTTCGC TTTCCCCCTC CCTATTGCCA 5461 CGGCGGAACT CATCGCCGCC TGCCTTGCCC GCTGCTGGAC AGGGGCTCGG CTGTTGGGCA 5521 CTGACAATTC CGTGGTGTTG TCGGGGAAGC TGACGTCCTT TCCATGGCTG CTCGCCTGTG 5581 TTGCCACCTG GATTCTGCGC GGGACGTCCT TCTGCTACGT CCCTTCGGCC CTCAATCCAG 5641 CGGACCTTCC TTCCCGCGGC CTGCTGCCGG CTCTGCGGCC TCTTCCGCGT CTTCGAGATC 5701 TGCCTCGACT GTGCCTTCTA GTTGCCAGCC ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC 5761 CTTGACCCTG GAAGGTGCCA CTCCCACTGT CCTTTCCTAA TAAAATGAGG AAATTGCATC 5821 GCATTGTCTG AGTAGGTGTC ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG 5881 GGAGGATTGG GAAGACAATA GCAGGCATGC TGGGGACTCG AGTTAAGGGC GAATTGCCGA 5941 TTAGGATCTT CCTAGAGCAT GGCTACGTAG ATAAGTAGCA TGGCGGGTTA ATCATTAACT 6001 ACAAGGAACC CCTAGTGATG GAGTTGGCCA CTCCCTCTCT GCGCGCTCGC TCGCTCACTG 6061 AGGCCGGGCG ACCAAAGGTC GCCCGACGCC CGGGCTTTGC CCGGGCGGCC TCAGTGAGCG 6121 AGCGAGCGCG CAGCCTTAAT TAAGAGCTCA TGGCGCGCCT AGGCCTTGAC GGCCTTCCGC 6181 CAATTCGCCC TATAGTGAGT CGTATTACGT CGCGCTCACT GGCCGTCGTT TTACAACGTC 6241 GTGACTGGGA AAACCCTGGC GTTACCCAAC TTAATCGCCT TGCAGCACAT CCCCCTTTCG 6301 CCAGCTGGCG TAATAGCGAA GAGGCCCGCA CCGAAACGCC CTTCCCAACA GTTGCGCAGC 6361 CTGAATGGCG AATGGGAGCG CCCTGTAGCG GCCACTCAAC CCTATCTCGG TCTATTCTTT 6421 TGATTTATAA GGGATTTTGC CGATTTCGGC CTATTGGTTA AAAAATGAGC TGATTTAACA 6481 AAAATTTAAC GCGAATTTTA ACAAAATATT AACGCTTACA ATTTAG //