LOCUS GLB1-pGEc2-DEST 7252 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6730..6859) /note="3'-ITR" misc_feature complement(6428..6642) /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 5880..6421 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 5059..5081 /note="attB2" misc_feature 5859..5864 /note="BstB1 site" misc_feature 5082..5777 /note="Super GFP" misc_feature 5778..5828 /note="AviTag" misc_feature 5829..5852 /note="His Tag" PCR_frame 3002..5059 /note="Reading Frame" /set="Set 1" source 3006..5036 /dnas_title="GLB1-pGEc2-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..7252,1..2991) /source="pGEc2-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 TATGAGCAAG GGCGAGGAAC 5101 TGTTCACCGG CGTGGTGCCC ATCCTGGTGG AACTGGATGG CGACGTGAAC GGCCACAAGT 5161 TCTCTGTGCG GGGAGAGGGC GAAGGCGACG CCACAAATGG CAAGCTGACC CTGAAGTTCA 5221 TCTGCACCAC CGGCAAGCTG CCCGTGCCTT GGCCTACCCT CGTGACCACA CTGACCTACG 5281 GCGTGCAGTG CTTCAGCAGA TACCCCGACC ATATGAAGCG GCACGACTTC TTCAAGAGCG 5341 CCATGCCCGA GGGCTACGTG CAGGAACGGA CCATCAGCTT CAAGGACGAC GGCACCTACA 5401 AGACCAGAGC CGAAGTGAAG TTCGAGGGCG ACACCCTCGT GAACCGGATC GAGCTGAAGG 5461 GCATCGATTT CAAAGAGGAC GGCAACATCC TGGGCCACAA GCTGGAGTAC AACTTCAACA 5521 GCCACAACGT GTACATCACC GCCGACAAGC AGAAGAACGG CATCAAGGCC AACTTCAAGA 5581 TCCGGCACAA CGTGGAAGAT GGCAGCGTGC AGCTGGCCGA CCACTACCAG CAGAACACCC 5641 CCATCGGAGA TGGCCCCGTG CTGCTGCCCG ACAACCACTA CCTGAGCACC CAGAGCGTGC 5701 TGAGCAAGGA CCCCAACGAG AAGCGGGACC ACATGGTGCT GCTGGAATTC GTGACCGCCG 5761 CTGGCATCAC CCACGGCATG AGCGGCCTGA ACGACATCTT CGAGGCCCAG AAAATCGAGT 5821 GGCACGAGCA CCACCATCAT CACCATCACC ACTGATGATT CGAAAAATAA TAAGGATCCA 5881 ATCAACCTCT GGATTACAAA ATTTGTGAAA GATTGACTGG TATTCTTAAC TATGTTGCTC 5941 CTTTTACGCT ATGTGGATAC GCTGCTTTAA TGCCTTTGTA TCATGCTATT GCTTCCCGTA 6001 TGGCTTTCAT TTTCTCCTCC TTGTATAAAT CCTGGTTGCT GTCTCTTTAT GAGGAGTTGT 6061 GGCCCGTTGT CAGGCAACGT GGCGTGGTGT GCACTGTGTT TGCTGACGCA ACCCCCACTG 6121 GTTGGGGCAT TGCCACCACC TGTCAGCTCC TTTCCGGGAC TTTCGCTTTC CCCCTCCCTA 6181 TTGCCACGGC GGAACTCATC GCCGCCTGCC TTGCCCGCTG CTGGACAGGG GCTCGGCTGT 6241 TGGGCACTGA CAATTCCGTG GTGTTGTCGG GGAAGCTGAC GTCCTTTCCA TGGCTGCTCG 6301 CCTGTGTTGC CACCTGGATT CTGCGCGGGA CGTCCTTCTG CTACGTCCCT TCGGCCCTCA 6361 ATCCAGCGGA CCTTCCTTCC CGCGGCCTGC TGCCGGCTCT GCGGCCTCTT CCGCGTCTTC 6421 GAGATCTGCC TCGACTGTGC CTTCTAGTTG CCAGCCATCT GTTGTTTGCC CCTCCCCCGT 6481 GCCTTCCTTG ACCCTGGAAG GTGCCACTCC CACTGTCCTT TCCTAATAAA ATGAGGAAAT 6541 TGCATCGCAT TGTCTGAGTA GGTGTCATTC TATTCTGGGG GGTGGGGTGG GGCAGGACAG 6601 CAAGGGGGAG GATTGGGAAG ACAATAGCAG GCATGCTGGG GACTCGAGTT AAGGGCGAAT 6661 TGCCGATTAG GATCTTCCTA GAGCATGGCT ACGTAGATAA GTAGCATGGC GGGTTAATCA 6721 TTAACTACAA GGAACCCCTA GTGATGGAGT TGGCCACTCC CTCTCTGCGC GCTCGCTCGC 6781 TCACTGAGGC CGGGCGACCA AAGGTCGCCC GACGCCCGGG CTTTGCCCGG GCGGCCTCAG 6841 TGAGCGAGCG AGCGCGCAGC CTTAATTAAG AGCTCATGGC GCGCCTAGGC CTTGACGGCC 6901 TTCCGCCAAT TCGCCCTATA GTGAGTCGTA TTACGTCGCG CTCACTGGCC GTCGTTTTAC 6961 AACGTCGTGA CTGGGAAAAC CCTGGCGTTA CCCAACTTAA TCGCCTTGCA GCACATCCCC 7021 CTTTCGCCAG CTGGCGTAAT AGCGAAGAGG CCCGCACCGA AACGCCCTTC CCAACAGTTG 7081 CGCAGCCTGA ATGGCGAATG GGAGCGCCCT GTAGCGGCCA CTCAACCCTA TCTCGGTCTA 7141 TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT TGGTTAAAAA ATGAGCTGAT 7201 TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG CTTACAATTT AG //