LOCUS HEXB-pGEc2-DEST 6889 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6367..6496) /note="3'-ITR" misc_feature complement(6065..6279) /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 5517..6058 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 4696..4718 /note="attB2" misc_feature 5496..5501 /note="BstB1 site" misc_feature 4719..5414 /note="Super GFP" misc_feature 5415..5465 /note="AviTag" misc_feature 5466..5489 /note="His Tag" PCR_frame 3002..4696 /note="Reading Frame" /set="Set 1" source 3006..4673 /dnas_title="HEXB-pGEc2-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:1725 IMAGE:2967035" /tissue_type="Skin, melanotic melanoma." /clone_lib="NIH_MGC_20" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 3006..4673 /dnas_title="HEXB" /gene="HEXB" /gene_synonym="ENC-1AS" /db_xref="GeneID:3074" /db_xref="HGNC:4879" /db_xref="MIM:606873" CDS 3006..4673 /dnas_title="hexosaminidase B (beta polypeptide)" /gene="HEXB" /gene_synonym="ENC-1AS" /codon_start=1 /product="hexosaminidase B (beta polypeptide)" /protein_id="AAH17378.1" /db_xref="GI:16924217" /db_xref="GeneID:3074" /db_xref="HGNC:4879" /db_xref="MIM:606873" /translation="MELCGLGLPRPPMLLALLLATLLAAMLALLTQVALVVQVAEAAR APSVSAKPGPALWPLPLSVKMTPNLLHLAPENFYISHSPNSTAGPSCTLLEEAFRRYH GYIFGFYKWHHEPAEFQAKTQVQQLLVSITLQSECDAFPNISSDESYTLLVKEPVAVL KANRVWGALRGLETFSQLVYQDSYGTFTINESTIIDSPRFSHRGILIDTSRHYLPVKI ILKTLDAMAFNKFNVLHWHIVDDQSFPYQSITFPELSNKGSYSLSHVYTPNDVRMVIE YARLRGIRVLPEFDTPGHTLSWGKGQKDLLTPCYSRQNKLDSFGPINPTLNTTYSFLT TFFKEISEVFPDQFIHLGGDEVEFKCWESNPKIQDFMRQKGFGTDFKKLESFYIQKVL DIIATINKGSIVWQEVFDDKAKLAPGTIVEVWKDSAYPEELSRVTASGFPVILSAPWY LDLISYGQDWRKYYKVEPLDFGGTQKQKQLFIGGEACLWGEYVDATNLTPRLWPRASA VGERLWSSKDVRDMDDAYDRLTRHRCRMVERGIAAQPLYAGYCNHENM" misc_difference 3190..3190 /dnas_title="HEXB" /gene="HEXB" /gene_synonym="ENC-1AS" /note="'C' in cDNA is 'T' in the human genome; amino acid difference: 'S' in cDNA, 'L' in the human genome. The chimpanzee genome agrees with the cDNA sequence, suggesting that this difference is unlikely to be due to an artifact." misc_feature 3006..4673 /note="HEXB coding region" misc_feature 4674..4694 /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..4703) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..4703) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(4704..6889,1..2991) /source="pGEc2-DEST" /type="Destination Vector" clone 2992..4703 /source="HEXB-Cterm-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 TCGCCatgga gctgtgcggg ctggggctgc cccggccgcc catgctgctg gcgctgctgt 3061 tggcgacact gctggcggcg atgttggcgc tgctgactca ggtggcgctg gtggtgcagg 3121 tggcggaggc ggctcgggcc ccgagcgtct cggccaagcc ggggccggcg ctgtggcccc 3181 tgccgctctc ggtgaagatg accccgaacc tgctgcatct cgccccggag aacttctaca 3241 tcagccacag ccccaattcc acggcgggcc cctcctgcac cctgctggag gaagcgtttc 3301 gacgatatca tggctatatt tttggtttct acaagtggca tcatgaacct gctgaattcc 3361 aggctaaaac ccaggttcag caacttcttg tctcaatcac ccttcagtca gagtgtgatg 3421 ctttccccaa catatcttca gatgagtctt atactttact tgtgaaagaa ccagtggctg 3481 tccttaaggc caacagagtt tggggagcat tacgaggttt agagaccttt agccagttag 3541 tttatcaaga ttcttatgga actttcacca tcaatgaatc caccattatt gattctccaa 3601 ggttttctca cagaggaatt ttgattgata catccagaca ttatctgcca gttaagatta 3661 ttcttaaaac tctggatgcc atggctttta ataagtttaa tgttcttcac tggcacatag 3721 ttgatgacca gtctttccca tatcagagca tcacttttcc tgagttaagc aataaaggaa 3781 gctattcttt gtctcatgtt tatacaccaa atgatgtccg tatggtgatt gaatatgcca 3841 gattacgagg aattcgagtc ctgccagaat ttgatacccc tgggcataca ctatcttggg 3901 gaaaaggtca gaaagacctc ctgactccat gttacagtag acaaaacaag ttggactctt 3961 ttggacctat aaaccctact ctgaatacaa catacagctt ccttactaca tttttcaaag 4021 aaattagtga ggtgtttcca gatcaattca ttcatttggg aggagatgaa gtggaattta 4081 aatgttggga atcaaatcca aaaattcaag atttcatgag gcaaaaaggc tttggcacag 4141 attttaagaa actagaatct ttctacattc aaaaggtttt ggatattatt gcaaccataa 4201 acaagggatc cattgtctgg caggaggttt ttgatgataa agcaaagctt gcgccgggca 4261 caatagttga agtatggaaa gacagcgcat atcctgagga actcagtaga gtcacagcat 4321 ctggcttccc tgtaatcctt tctgctcctt ggtacttaga tttgattagc tatggacaag 4381 attggaggaa atactataaa gtggaacctc ttgattttgg cggtactcag aaacagaaac 4441 aacttttcat tggtggagaa gcttgtctat ggggagaata tgtggatgca actaacctca 4501 ctccaagatt atggcctcgg gcaagtgctg ttggtgagag actctggagt tccaaagatg 4561 tcagagatat ggatgacgcc tatgacagac tgacaaggca ccgctgcagg atggtcgaac 4621 gtggaatagc tgcacaacct ctttatgctg gatattgtaa ccatgagaac atggaaaact 4681 tgtactttca aggctCCAGC TTTCTTGTAC AAAGTGGTAT GAGCAAGGGC GAGGAACTGT 4741 TCACCGGCGT GGTGCCCATC CTGGTGGAAC TGGATGGCGA CGTGAACGGC CACAAGTTCT 4801 CTGTGCGGGG AGAGGGCGAA GGCGACGCCA CAAATGGCAA GCTGACCCTG AAGTTCATCT 4861 GCACCACCGG CAAGCTGCCC GTGCCTTGGC CTACCCTCGT GACCACACTG ACCTACGGCG 4921 TGCAGTGCTT CAGCAGATAC CCCGACCATA TGAAGCGGCA CGACTTCTTC AAGAGCGCCA 4981 TGCCCGAGGG CTACGTGCAG GAACGGACCA TCAGCTTCAA GGACGACGGC ACCTACAAGA 5041 CCAGAGCCGA AGTGAAGTTC GAGGGCGACA CCCTCGTGAA CCGGATCGAG CTGAAGGGCA 5101 TCGATTTCAA AGAGGACGGC AACATCCTGG GCCACAAGCT GGAGTACAAC TTCAACAGCC 5161 ACAACGTGTA CATCACCGCC GACAAGCAGA AGAACGGCAT CAAGGCCAAC TTCAAGATCC 5221 GGCACAACGT GGAAGATGGC AGCGTGCAGC TGGCCGACCA CTACCAGCAG AACACCCCCA 5281 TCGGAGATGG CCCCGTGCTG CTGCCCGACA ACCACTACCT GAGCACCCAG AGCGTGCTGA 5341 GCAAGGACCC CAACGAGAAG CGGGACCACA TGGTGCTGCT GGAATTCGTG ACCGCCGCTG 5401 GCATCACCCA CGGCATGAGC GGCCTGAACG ACATCTTCGA GGCCCAGAAA ATCGAGTGGC 5461 ACGAGCACCA CCATCATCAC CATCACCACT GATGATTCGA AAAATAATAA GGATCCAATC 5521 AACCTCTGGA TTACAAAATT TGTGAAAGAT TGACTGGTAT TCTTAACTAT GTTGCTCCTT 5581 TTACGCTATG TGGATACGCT GCTTTAATGC CTTTGTATCA TGCTATTGCT TCCCGTATGG 5641 CTTTCATTTT CTCCTCCTTG TATAAATCCT GGTTGCTGTC TCTTTATGAG GAGTTGTGGC 5701 CCGTTGTCAG GCAACGTGGC GTGGTGTGCA CTGTGTTTGC TGACGCAACC CCCACTGGTT 5761 GGGGCATTGC CACCACCTGT CAGCTCCTTT CCGGGACTTT CGCTTTCCCC CTCCCTATTG 5821 CCACGGCGGA ACTCATCGCC GCCTGCCTTG CCCGCTGCTG GACAGGGGCT CGGCTGTTGG 5881 GCACTGACAA TTCCGTGGTG TTGTCGGGGA AGCTGACGTC CTTTCCATGG CTGCTCGCCT 5941 GTGTTGCCAC CTGGATTCTG CGCGGGACGT CCTTCTGCTA CGTCCCTTCG GCCCTCAATC 6001 CAGCGGACCT TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG GCCTCTTCCG CGTCTTCGAG 6061 ATCTGCCTCG ACTGTGCCTT CTAGTTGCCA GCCATCTGTT GTTTGCCCCT CCCCCGTGCC 6121 TTCCTTGACC CTGGAAGGTG CCACTCCCAC TGTCCTTTCC TAATAAAATG AGGAAATTGC 6181 ATCGCATTGT CTGAGTAGGT GTCATTCTAT TCTGGGGGGT GGGGTGGGGC AGGACAGCAA 6241 GGGGGAGGAT TGGGAAGACA ATAGCAGGCA TGCTGGGGAC TCGAGTTAAG GGCGAATTGC 6301 CGATTAGGAT CTTCCTAGAG CATGGCTACG TAGATAAGTA GCATGGCGGG TTAATCATTA 6361 ACTACAAGGA ACCCCTAGTG ATGGAGTTGG CCACTCCCTC TCTGCGCGCT CGCTCGCTCA 6421 CTGAGGCCGG GCGACCAAAG GTCGCCCGAC GCCCGGGCTT TGCCCGGGCG GCCTCAGTGA 6481 GCGAGCGAGC GCGCAGCCTT AATTAAGAGC TCATGGCGCG CCTAGGCCTT GACGGCCTTC 6541 CGCCAATTCG CCCTATAGTG AGTCGTATTA CGTCGCGCTC ACTGGCCGTC GTTTTACAAC 6601 GTCGTGACTG GGAAAACCCT GGCGTTACCC AACTTAATCG CCTTGCAGCA CATCCCCCTT 6661 TCGCCAGCTG GCGTAATAGC GAAGAGGCCC GCACCGAAAC GCCCTTCCCA ACAGTTGCGC 6721 AGCCTGAATG GCGAATGGGA GCGCCCTGTA GCGGCCACTC AACCCTATCT CGGTCTATTC 6781 TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG AGCTGATTTA 6841 ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTAG //