LOCUS PYGB-pGEc1-DEST 7024 bp DNA circular 22-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6502..6631) /note="3'-ITR" misc_feature complement(6200..6414) /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 5652..6193 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_recomb 5557..5579 /label=attB2 /note="attB2 site" misc_feature 5589..5612 /note="8xHis Tag" misc_feature 5616..5639 /note="StrepTag" PCR_frame 3002..5557 /note="Reading Frame" /set="Set 1" source 3006..5534 /dnas_title="PYGB-pGEc1-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:9213 IMAGE:3877951" /tissue_type="Lung, large cell carcinoma" /clone_lib="NIH_MGC_68" /lab_host="DH10B" /note="Vector: pCMV-SPORT6" gene 3006..5534 /dnas_title="PYGB" /gene="PYGB" /gene_synonym="MGC9213" /db_xref="GeneID:5834" /db_xref="HGNC:9723" /db_xref="MIM:138550" CDS 3006..5534 /dnas_title="phosphorylase, glycogen; brain" /gene="PYGB" /gene_synonym="MGC9213" /codon_start=1 /product="phosphorylase, glycogen; brain" /protein_id="AAH17045.1" /db_xref="GI:16877586" /db_xref="GeneID:5834" /db_xref="HGNC:9723" /db_xref="MIM:138550" /translation="MAKPLTDSEKRKQISVRGLAGLGDVAEVRKSFNRHLHFTLVKDR NVATPRDYFFALAHTVRDHLVGRWIRTQQHYYERDPKRIYYLSLEFYMGRTLQNTMVN LGLQNACDEAIYQLGLDLEELEEIEEDAGLGNGGLGRLAACFLDSMATLGLAAYGYGI RYEFGIFNQKIVNGWQVEEADDWLRYGNPWEKARPEYMLPVHFYGRVEHTPDGVKWLD TQVVLAMPYDTPVPGYKNNTVNTMRLWSAKAPNDFKLQDFNVGDYIEAVLDRNLAENI SRVLYPNDNFFEGKELRLKQEYFVVAATLQDIIRRFKSSKFGCRDPVRTCFETFPDKV AIQLNDTHPALSIPELMRILVDVEKVDWDKAWEITKKTCAYTNHTVLPEALERWPVSM FEKLLPRHLEIIYAINQRHLDHVAALFPGDVDRLRRMSVIEEGDCKRINMAHLCVIGS HAVNGVARIHSEIVKQSVFKDFYELEPEKFQNKTNGITPRRWLLLCNPGLADTIVEKI GEEFLTDLSQLKKLLPLVSDEVFIRDVAKVKQENKLKFSAFLEKEYKVKINPSSMFDV HVKRIHEYKRQLLNCLHVVTLYNRIKRDPAKAFVPRTVMIGGKAAPGYHMAKLIIKLV TSIGDVVNHDPVVGDRLKVIFLENYRVSLAEKVIPAADLSQQISTAGTEASGTGNMKF MLNGALTIGTMDGANVEMAEEAGAENLFIFGLRVEDVEALDRKGYNAREYYDHLPELK QAVDQISSGFFSPKEPDCFKDIVNMLMHHDRFKVFADYEAYMQCQAQVDQLYRNPKEW TKKVIRNIACSGKFSSDRTITEYAREIWGVEPSDLQIPPPNIPRD" misc_feature 3006..5534 /note="PYGB coding region" misc_feature 5535..5555 /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..5564) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..5564) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(5565..7024,1..2991) /source="pGEc1-DEST" /type="Destination Vector" clone 2992..5564 /source="PYGB-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 TCGCCatggc gaagccgctg acggacagcg agaagcggaa gcagatcagc gtgcgcggcc 3061 tggcggggct aggcgacgtg gccgaggtgc ggaagagctt caaccggcac ttgcacttca 3121 cgctggtcaa ggaccgcaat gtggccacgc cccgcgacta cttcttcgcg ctggcgcaca 3181 cggtgcgcga ccacctcgtg ggccgctgga tccgcacgca gcagcactac tacgagcgcg 3241 accccaagcg catttattat ctttccctgg aattctacat gggtcgcacg ctgcagaaca 3301 cgatggtgaa cctgggcctt cagaatgcct gcgatgaagc catctatcag ttggggttag 3361 acttggagga actcgaggag atagaagaag atgctggcct tgggaatgga ggcctgggga 3421 ggctggcagc gtgtttcctt gactcaatgg ctaccttggg cctggcagca tacggctatg 3481 gaatccgcta tgaatttggg atttttaacc agaagattgt caatggctgg caggtagagg 3541 aggccgatga ctggctgcgc tacggcaacc cctgggagaa agcgcggcct gagtatatgc 3601 ttcccgtgca cttctacgga cgcgtggagc acacccccga cggcgtgaag tggctggaca 3661 cacaggtggt gctggccatg ccctacgaca ccccagtgcc cggctacaag aacaacaccg 3721 tcaacaccat gcggctgtgg tccgccaagg ctcccaacga cttcaagctg caggacttca 3781 acgtgggaga ctacatcgag gcggtcctgg accggaactt ggctgagaac atctccaggg 3841 tcctgtatcc aaatgataac ttctttgagg ggaaggagct gcggctgaag caggagtact 3901 tcgtggtggc cgccacgctc caggacatca tccgccgctt caagtcgtcc aagttcggct 3961 gccgggaccc tgtgagaacc tgtttcgaga cgttcccaga caaggtggcc atccagctga 4021 acgacaccca ccccgccctc tccatccctg agctcatgcg gatcctggtg gacgtggaga 4081 aggtggactg ggacaaggcc tgggaaatca cgaagaagac ctgtgcatac accaaccaca 4141 ctgtgctgcc tgaggccttg gagcgctggc ccgtgtccat gtttgagaag ctgctgccgc 4201 ggcacctgga gataatctat gccatcaacc agcggcacct ggaccacgtg gccgcgctgt 4261 ttcccggcga tgtggaccgc ctgcgcagga tgtctgtgat cgaggagggg gactgcaagc 4321 ggatcaacat ggcccacctg tgtgtgattg ggtcccatgc tgtcaatggt gtggcgagga 4381 tccactcgga gatcgtgaaa cagtcggtct ttaaggattt ttatgaactg gagccagaga 4441 agttccagaa taagaccaat ggcatcaccc cccgccggtg gctgctgctg tgcaacccgg 4501 ggctggccga taccatcgtg gagaaaattg gggaggagtt cctgactgac ctgagccagc 4561 tgaagaagct gctgccgctg gtcagtgacg aggtgttcat cagggacgtg gccaaggtca 4621 aacaggagaa caagctcaag ttctcggcct tcctggagaa ggagtacaag gtgaagatca 4681 acccctcctc catgttcgat gtgcatgtga agaggatcca cgagtacaag cggcagctgc 4741 tcaactgcct gcacgtcgtc accctgtaca atcgaatcaa gagagacccg gccaaggctt 4801 ttgtgcccag gactgttatg attgggggca aggcagcgcc cggttaccac atggccaagc 4861 tgatcatcaa gttggtcacc tccatcggcg acgtcgtcaa tcatgaccca gttgtgggtg 4921 acaggttgaa agtgatcttc ctggagaact accgtgtgtc cttggctgag aaagtgatcc 4981 cggccgctga tctgtcgcag cagatctcca ctgcaggcac cgaggcctca ggcacaggca 5041 acatgaagtt catgctcaac ggggccctca ccatcggcac catggacggc gccaacgtgg 5101 agatggccga ggaggccggg gccgagaacc tcttcatctt cggcctgcgg gtggaggatg 5161 tcgaggcctt ggaccggaaa gggtacaatg ccagggagta ctacgaccac ctgcccgagc 5221 tgaagcaggc cgtggaccag atcagcagtg gctttttttc tcccaaggag ccagactgct 5281 tcaaggacat cgtgaacatg ctgatgcacc atgacaggtt caaggtgttt gcagactatg 5341 aagcctacat gcagtgccag gcacaggtgg accagctgta ccggaacccc aaggagtgga 5401 ccaagaaggt catcaggaac atcgcctgct cgggcaagtt ctccagtgac cggaccatca 5461 cggagtatgc acgggagatc tggggtgtgg agccctccga cctgcagatc ccgcccccca 5521 acatcccccg ggacgaaaac ttgtactttc aaggctCCAG CTTTCTTGTA CAAAGTGGTG 5581 ATGGCCGCCA TCATCATCAT CATCATCATC ATGCGTGGAG CCACCCGCAG TTCGAAAAAT 5641 AATAAGGATC CAATCAACCT CTGGATTACA AAATTTGTGA AAGATTGACT GGTATTCTTA 5701 ACTATGTTGC TCCTTTTACG CTATGTGGAT ACGCTGCTTT AATGCCTTTG TATCATGCTA 5761 TTGCTTCCCG TATGGCTTTC ATTTTCTCCT CCTTGTATAA ATCCTGGTTG CTGTCTCTTT 5821 ATGAGGAGTT GTGGCCCGTT GTCAGGCAAC GTGGCGTGGT GTGCACTGTG TTTGCTGACG 5881 CAACCCCCAC TGGTTGGGGC ATTGCCACCA CCTGTCAGCT CCTTTCCGGG ACTTTCGCTT 5941 TCCCCCTCCC TATTGCCACG GCGGAACTCA TCGCCGCCTG CCTTGCCCGC TGCTGGACAG 6001 GGGCTCGGCT GTTGGGCACT GACAATTCCG TGGTGTTGTC GGGGAAGCTG ACGTCCTTTC 6061 CATGGCTGCT CGCCTGTGTT GCCACCTGGA TTCTGCGCGG GACGTCCTTC TGCTACGTCC 6121 CTTCGGCCCT CAATCCAGCG GACCTTCCTT CCCGCGGCCT GCTGCCGGCT CTGCGGCCTC 6181 TTCCGCGTCT TCGAGATCTG CCTCGACTGT GCCTTCTAGT TGCCAGCCAT CTGTTGTTTG 6241 CCCCTCCCCC GTGCCTTCCT TGACCCTGGA AGGTGCCACT CCCACTGTCC TTTCCTAATA 6301 AAATGAGGAA ATTGCATCGC ATTGTCTGAG TAGGTGTCAT TCTATTCTGG GGGGTGGGGT 6361 GGGGCAGGAC AGCAAGGGGG AGGATTGGGA AGACAATAGC AGGCATGCTG GGGACTCGAG 6421 TTAAGGGCGA ATTGCCGATT AGGATCTTCC TAGAGCATGG CTACGTAGAT AAGTAGCATG 6481 GCGGGTTAAT CATTAACTAC AAGGAACCCC TAGTGATGGA GTTGGCCACT CCCTCTCTGC 6541 GCGCTCGCTC GCTCACTGAG GCCGGGCGAC CAAAGGTCGC CCGACGCCCG GGCTTTGCCC 6601 GGGCGGCCTC AGTGAGCGAG CGAGCGCGCA GCCTTAATTA AGAGCTCATG GCGCGCCTAG 6661 GCCTTGACGG CCTTCCGCCA ATTCGCCCTA TAGTGAGTCG TATTACGTCG CGCTCACTGG 6721 CCGTCGTTTT ACAACGTCGT GACTGGGAAA ACCCTGGCGT TACCCAACTT AATCGCCTTG 6781 CAGCACATCC CCCTTTCGCC AGCTGGCGTA ATAGCGAAGA GGCCCGCACC GAAACGCCCT 6841 TCCCAACAGT TGCGCAGCCT GAATGGCGAA TGGGAGCGCC CTGTAGCGGC CACTCAACCC 6901 TATCTCGGTC TATTCTTTTG ATTTATAAGG GATTTTGCCG ATTTCGGCCT ATTGGTTAAA 6961 AAATGAGCTG ATTTAACAAA AATTTAACGC GAATTTTAAC AAAATATTAA CGCTTACAAT 7021 TTAG //