LOCUS GBE1-pGEn3-DEST 8112 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 6740..7281 /note="WPRE" misc_recomb 6702..6724 /label=attB2 /note="attB2 site" misc_feature complement(7288..7502) /note="BGH pA" misc_feature complement(7590..7719) /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..6701 /dnas_title="GBE1-pGEn3-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:20071 IMAGE:4574938" /tissue_type="Primary B-Cells from Tonsils" /clone_lib="NIH_MGC_48" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 4593..6701 /dnas_title="GBE1" /gene="GBE1" /gene_synonym="GBE" /db_xref="GeneID:2632" /db_xref="HGNC:4180" /db_xref="MIM:607839" CDS 4593..6701 /dnas_title="glucan (1,4-alpha-), branching enzyme 1" /gene="GBE1" /gene_synonym="GBE" /codon_start=1 /product="glucan (1,4-alpha-), branching enzyme 1" /protein_id="AAH12098.1" /db_xref="GI:15082371" /db_xref="GeneID:2632" /db_xref="HGNC:4180" /db_xref="MIM:607839" /translation="MAAPMTPAARPEDYEAALNAALADVPELARLLEIDPYLKPYAVD FQRRYKQFSQILKNIGENEGGIDKFSRGYESFGVHRCADGGLYCKEWAPGAEGVFLTG DFNGWNPFSYPYKKLDYGKWELYIPPKQNKSVLVPHGSKLKVVITSKSGEILYRISPW AKYVVREGDNVNYDWIHWDPEHSYEFKHSRPKKPRSLRIYESHVGISSHEGKVASYKH FTCNVLPRIKGLGYNCIQLMAIMEHAYYASFGYQITSFFAASSRYGSPEELQELVDTA HSMGIIVLLDVVHSHASKNSADGLNMFDGTDSCYFHSGPRGTHDLWDSRLFAYSSWEV LRFLLSNIRWWLEEYRFDGFRFDGVTSMLYHHHGVGQGFSGDYSEYFGLQVDEDALTY LMLANHLVHTLCPDSITIAEDVSGMPALCSPISQGGGGFDYRLAMAIPDKWIQLLKEF KDEDWNMGDIVYTLTNRRYLEKCIAYAESHDQALVGDKSLAFWLMDAEMYTNMSVLTP FTPVIDRGIQLHKMIRLITHGLGGEGYLNFMGNEFGHPEWLDFPRKGNNESYHYARRQ FHLTDDDLLRYKFLNNFDRDMNRLEERYGWLAAPQAYVSEKHEGNKIIAFERAGLLFI FNFHPSKSYTDYRVGTALPGKFKIVLDSDAAEYGGHQRLDHSTDFFSEAFEHNGRPYS LLVYIPSRVALILQNVDLPN" misc_feature 4593..6701 /note="GBE1 coding region" misc_difference 5385..5385 /dnas_title="GBE1" /gene="GBE1" /gene_synonym="GBE" /note="'T' in cDNA is 'A' in the human genome; amino acid difference: 'S' in cDNA, 'T' in the human genome. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." misc_difference 5592..5592 /dnas_title="GBE1" /gene="GBE1" /gene_synonym="GBE" /note="'G' in cDNA is 'A' in the human genome; amino acid difference: 'V' in cDNA, 'I' in the human genome. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." vector join(6710..8112,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..6709 /source="GBE1-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 gcatggcggc tccgatgact cccgcggctc 4621 ggcccgagga ctacgaggcg gcgctcaatg ccgccctggc tgacgtgccc gaactggcca 4681 gactcctgga gatcgacccg tacttgaagc cctacgccgt ggacttccag cgcaggtata 4741 agcagtttag ccaaattttg aagaacattg gagaaaatga aggtggtatt gataagtttt 4801 ccagaggcta tgaatcattt ggcgtccaca gatgtgctga tggtggttta tactgcaaag 4861 aatgggcccc gggagcagaa ggagtttttc ttactggaga ttttaatggt tggaatccat 4921 tttcgtaccc atacaaaaaa ctggattatg gaaaatggga gctgtatatc ccaccaaagc 4981 agaataaatc tgtactcgtg cctcatggat ccaaattaaa ggtagttatt actagtaaaa 5041 gcggagagat cttgtatcgt atttcaccgt gggcaaagta tgtggttcgt gaaggtgata 5101 atgtgaatta tgattggata cactgggatc cagaacactc atatgagttt aagcattcca 5161 gaccaaagaa gccacggagt ctaagaattt atgaatctca tgtgggaatt tcttcccatg 5221 aaggaaaagt agcttcttat aaacatttta catgcaatgt actaccaaga atcaaaggcc 5281 ttggatacaa ctgcattcag ttgatggcaa tcatggagca tgcttactat gccagctttg 5341 gttaccaaat cacaagcttc tttgcagctt ccagccgtta tggatcacct gaagagctac 5401 aagaactggt agacacagct cattccatgg gtatcatagt cctcttagat gtggtacaca 5461 gccatgcttc aaaaaattca gcagatggat tgaatatgtt tgatgggaca gattcctgtt 5521 attttcattc tggacctaga gggactcatg atctttggga tagcagattg tttgcctact 5581 ccagctggga agttttaaga ttccttctgt caaacataag atggtggttg gaagaatatc 5641 gctttgatgg atttcgtttt gatggtgtta cgtccatgct ttatcatcac catggagtgg 5701 gtcaaggttt ctcaggtgat tacagtgaat atttcggact acaagtagat gaagatgcct 5761 tgacttacct catgttggca aatcatttgg ttcacacgct gtgtcccgat tctataacaa 5821 tagctgagga tgtatcagga atgccagctc tgtgctctcc aatttcccag ggagggggtg 5881 gttttgacta tcgactagcc atggcaattc cagataagtg gattcagcta cttaaagagt 5941 ttaaagatga agactggaac atgggcgata tagtatacac gctcacaaac aggcgctacc 6001 ttgaaaagtg cattgcttat gcagagagcc atgatcaggc attggttggg gataagtcgc 6061 tggcattttg gttgatggat gccgaaatgt atacaaacat gagtgtcctg actcctttta 6121 ctccagttat tgatcgtgga atacagcttc ataaaatgat tcgactcatt acgcatgggc 6181 ttggtggaga aggctatctc aatttcatgg gtaatgaatt tgggcatcct gaatggttag 6241 acttcccaag aaaaggaaat aatgagagtt accattatgc caggcggcag tttcatttaa 6301 ctgacgacga ccttcttcgc tacaagttcc taaataattt tgacagggat atgaatagat 6361 tggaagaaag atatggttgg cttgcagctc cacaggccta cgtgagtgaa aaacatgaag 6421 gcaataagat cattgctttt gaaagagcag gtcttctttt cattttcaac ttccatccaa 6481 gcaagagcta cactgactac cgagttggaa cagcattgcc agggaaattc aaaattgtgc 6541 tagattcaga tgcagcggaa tatggagggc atcagagact ggaccacagc actgactttt 6601 tttctgaggc ttttgaacat aatgggcgtc cctattctct tttggtgtac attccaagca 6661 gagtggccct catccttcag aatgtggatc tgccgaattg aCCAGCTTTC TTGTACAAAG 6721 TGGTTGATAG CTTGGATCCA ATCAACCTCT GGATTACAAA ATTTGTGAAA GATTGACTGG 6781 TATTCTTAAC TATGTTGCTC CTTTTACGCT ATGTGGATAC GCTGCTTTAA TGCCTTTGTA 6841 TCATGCTATT GCTTCCCGTA TGGCTTTCAT TTTCTCCTCC TTGTATAAAT CCTGGTTGCT 6901 GTCTCTTTAT GAGGAGTTGT GGCCCGTTGT CAGGCAACGT GGCGTGGTGT GCACTGTGTT 6961 TGCTGACGCA ACCCCCACTG GTTGGGGCAT TGCCACCACC TGTCAGCTCC TTTCCGGGAC 7021 TTTCGCTTTC CCCCTCCCTA TTGCCACGGC GGAACTCATC GCCGCCTGCC TTGCCCGCTG 7081 CTGGACAGGG GCTCGGCTGT TGGGCACTGA CAATTCCGTG GTGTTGTCGG GGAAGCTGAC 7141 GTCCTTTCCA TGGCTGCTCG CCTGTGTTGC CACCTGGATT CTGCGCGGGA CGTCCTTCTG 7201 CTACGTCCCT TCGGCCCTCA ATCCAGCGGA CCTTCCTTCC CGCGGCCTGC TGCCGGCTCT 7261 GCGGCCTCTT CCGCGTCTTC GAGATCTGCC TCGACTGTGC CTTCTAGTTG CCAGCCATCT 7321 GTTGTTTGCC CCTCCCCCGT GCCTTCCTTG ACCCTGGAAG GTGCCACTCC CACTGTCCTT 7381 TCCTAATAAA ATGAGGAAAT TGCATCGCAT TGTCTGAGTA GGTGTCATTC TATTCTGGGG 7441 GGTGGGGTGG GGCAGGACAG CAAGGGGGAG GATTGGGAAG ACAATAGCAG GCATGCTGGG 7501 GACTCGAGTT AAGGGCGAAT TGCCGATTAG GATCTTCCTA GAGCATGGCT ACGTAGATAA 7561 GTAGCATGGC GGGTTAATCA TTAACTACAA GGAACCCCTA GTGATGGAGT TGGCCACTCC 7621 CTCTCTGCGC GCTCGCTCGC TCACTGAGGC CGGGCGACCA AAGGTCGCCC GACGCCCGGG 7681 CTTTGCCCGG GCGGCCTCAG TGAGCGAGCG AGCGCGCAGC CTTAATTAAG AGCTCATGGC 7741 GCGCCTAGGC CTTGACGGCC TTCCGCCAAT TCGCCCTATA GTGAGTCGTA TTACGTCGCG 7801 CTCACTGGCC GTCGTTTTAC AACGTCGTGA CTGGGAAAAC CCTGGCGTTA CCCAACTTAA 7861 TCGCCTTGCA GCACATCCCC CTTTCGCCAG CTGGCGTAAT AGCGAAGAGG CCCGCACCGA 7921 AACGCCCTTC CCAACAGTTG CGCAGCCTGA ATGGCGAATG GGAGCGCCCT GTAGCGGCCA 7981 CTCAACCCTA TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT 8041 TGGTTAAAAA ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG 8101 CTTACAATTT AG //