LOCUS PYGL-pGEc1-DEST 7033 bp DNA circular 22-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6511..6640) /note="3'-ITR" misc_feature complement(6209..6423) /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 5661..6202 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_recomb 5566..5588 /label=attB2 /note="attB2 site" misc_feature 5598..5621 /note="8xHis Tag" misc_feature 5625..5648 /note="StrepTag" PCR_frame 3002..5566 /note="Reading Frame" /set="Set 1" source 3006..5543 /dnas_title="PYGL-pGEc1-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:2000 IMAGE:2967205" /tissue_type="Skin, melanotic melanoma." /clone_lib="NIH_MGC_20" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 3006..5543 /dnas_title="PYGL" /gene="PYGL" /gene_synonym="GSD6" /db_xref="GeneID:5836" /db_xref="HGNC:9725" /db_xref="MIM:232700" CDS 3006..5543 /dnas_title="phosphorylase, glycogen, liver" /gene="PYGL" /gene_synonym="GSD6" /codon_start=1 /product="phosphorylase, glycogen, liver" /protein_id="AAH09895.3" /db_xref="GI:112180335" /db_xref="GeneID:5836" /db_xref="HGNC:9725" /db_xref="MIM:232700" /translation="MAKPLTDQEKRRQISIRGIVGVENVAELKKSFNRHLHFTLVKDR NVATTRDYYFALAHTVRDHLVGRWIRTQQHYYDKCPKRVYYLSLEFYMGRTLQNTMIN LGLQNACDEAIYQLGLDIEELEEIEEDAGLGNGGLGRLAACFLDSMATLGLAAYGYGI RYEYGIFNQKIRDGWQVEEADDWLRYGNPWEKSRPEFMLPVHFYGKVEHTNTGTKWID TQVVLALPYDTPVPGYMNNTVNTMRLWSARAPNDFNLRDFNVGDYIQAVLDRNLAENI SRVLYPNDNFFEGKELRLKQEYFVVAATLQDIIRRFKASKFGSTRGAGTVFDAFPDQV AIQLNDTHPALAIPELMRIFVDIEKLPWSKAWELTQKTFAYTNHTVLPEALERWPVDL VEKLLPRHLEIIYEINQKHLDRIVALFPKDVDRLRRMSLIEEEGSKRINMAHLCIVGS HAVNGVAKIHSDIVKTKVFKDFSELEPDKFQNKTNGITPRRWLLLCNPGLAELIAEKI GEDYVKDLSQLTKLHSFLGDDVFLRELAKVKQENKLKFSQFLETEYKVKINPSSMFDV QVKRIHEYKRQLLNCLHVITMYNRIKKDPKKLFVPRTVIIGGKAAPGYHMAKMIIKLI TSVADVVNNDPMVGSKLKVIFLENYRVSLAEKVIPATDLSEQISTAGTEASGTGNMKF LNGALTIGTMDGANVEMAEEAGEENLFIFGMRIDDVAALDKKGYEAKEYYEALPELKL VIDQIDNGFFSPKQPDLFKDIINMLFYHDRFKVFADYEAYVKCQDKVSQLYMNPKAWN TMVLKNIAASGKFSSDRTIKEYAQNIWNVEPSDLKISLSNESNKVNGN" misc_feature 3006..5543 /note="PYGL coding region" misc_difference 5051..5052 /dnas_title="PYGL" /gene="PYGL" /gene_synonym="GSD6" /note="3 bases in the human genome, ATG, are not found in cDNA." misc_feature 5544..5564 /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..5573) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..5573) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(5574..7033,1..2991) /source="pGEc1-DEST" /type="Destination Vector" clone 2992..5573 /source="PYGL-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 gaagcccctg acggaccagg agaagcggcg gcagatcagc atccgcggca 3061 tcgtgggcgt ggagaacgtg gcagagctga agaagagttt caaccggcac ctgcacttca 3121 cgctggtcaa ggaccgcaac gtggccacca cccgcgacta ctacttcgcg ctggcgcaca 3181 cggtgcgcga ccacctggtg gggcgctgga tccgcacgca gcagcactac tacgacaagt 3241 gccccaagag ggtatattac ctctctctgg aattttacat gggccgaaca ttacagaaca 3301 ccatgatcaa cctcggtctg caaaatgcct gtgatgaggc catttaccag cttggattgg 3361 atatagaaga gttagaagaa attgaagaag atgctggact tggcaatggt ggtcttggga 3421 gacttgctgc ctgcttcttg gattccatgg caaccctggg acttgcagcc tatggatacg 3481 gcattcggta tgaatatggg attttcaatc agaagatccg agatggatgg caggtagaag 3541 aagcagatga ttggctcaga tatggaaacc cttgggagaa gtcccgccca gaattcatgc 3601 tgcctgtgca cttctatgga aaagtagaac acaccaacac cgggaccaag tggattgaca 3661 ctcaagtggt cctggctctg ccatatgaca cccccgtgcc cggctacatg aataacactg 3721 tcaacaccat gcgcctctgg tctgctcggg caccaaatga ctttaacctc agagacttta 3781 atgttggaga ctacattcag gctgtgctgg accgaaacct ggccgagaac atctcccggg 3841 tcctctatcc caatgacaat ttttttgaag ggaaggagct aagattgaag caggaatact 3901 ttgtggtggc tgcaaccttg caagatatca tccgccgttt caaagcctcc aagtttggct 3961 ccacccgtgg tgcaggaact gtgtttgatg ccttcccgga tcaggtggcc atccagctga 4021 atgacactca ccctgcactc gcgatccctg agctgatgag gatttttgtg gatattgaaa 4081 aactgccctg gtccaaggca tgggagctca cccagaagac cttcgcctac accaaccaca 4141 cagtgctccc ggaagccctg gagcgctggc ccgtggacct ggtggagaag ctgctccctc 4201 gacatttgga aatcatttat gagataaatc agaagcattt agatagaatt gtggccttgt 4261 ttcctaaaga tgtggaccgt ctgagaagga tgtctctgat agaagaggaa ggaagcaaaa 4321 ggatcaacat ggcccatctc tgcattgtcg gttcccatgc tgtgaatggc gtggctaaaa 4381 tccactcaga catcgtgaag actaaagtat tcaaggactt cagtgagcta gaacctgaca 4441 agtttcagaa taaaaccaat gggatcactc caaggcgctg gctcctactc tgcaacccag 4501 gacttgcaga gctcatagca gagaaaattg gagaagacta tgtgaaagac ctgagccagc 4561 tgacgaagct ccacagcttc ctgggtgatg atgtcttcct ccgggaactc gccaaggtga 4621 agcaggagaa taagctgaag ttttctcagt tcctggagac ggagtacaaa gtgaagatca 4681 acccatcctc catgtttgat gtccaggtga agaggataca tgagtacaag cgacagctct 4741 tgaactgtct gcatgtgatc acgatgtaca accgcattaa gaaagaccct aagaagttat 4801 tcgtgccaag gacagttatc attggtggta aagctgcccc aggatatcac atggccaaaa 4861 tgatcataaa gctgatcact tcagtggcag atgtggtgaa caatgaccct atggttggaa 4921 gcaagttgaa agtcatcttc ttggagaact acagagtatc tcttgctgaa aaagtcattc 4981 cagccacaga tctgtcagag cagatttcca ctgcaggcac cgaagcctcg gggacaggca 5041 atatgaagtt cctaaatggg gccctaacta tcgggaccat ggatggggcc aatgtggaaa 5101 tggcagaaga agctggggaa gagaacctgt tcatctttgg catgaggata gatgatgtgg 5161 ctgctttgga caagaaaggg tacgaggcaa aagaatacta tgaggcactt ccagagctga 5221 agctggtcat tgatcaaatt gacaatggct ttttttctcc caagcagcct gacctcttca 5281 aagatatcat caacatgcta ttttatcatg acaggtttaa agtctttgca gactacgaag 5341 cctatgtcaa gtgtcaagat aaagtgagtc agctgtacat gaatccaaag gcctggaaca 5401 caatggtact caaaaacata gctgcctcgg ggaaattctc cagtgaccga acaattaaag 5461 aatatgccca aaacatctgg aacgtggaac cttcagatct aaagatttct ctatccaatg 5521 aatctaacaa agtcaatgga aatgaaaact tgtactttca aggctCCAGC TTTCTTGTAC 5581 AAAGTGGTGA TGGCCGCCAT CATCATCATC ATCATCATCA TGCGTGGAGC CACCCGCAGT 5641 TCGAAAAATA ATAAGGATCC AATCAACCTC TGGATTACAA AATTTGTGAA AGATTGACTG 5701 GTATTCTTAA CTATGTTGCT CCTTTTACGC TATGTGGATA CGCTGCTTTA ATGCCTTTGT 5761 ATCATGCTAT TGCTTCCCGT ATGGCTTTCA TTTTCTCCTC CTTGTATAAA TCCTGGTTGC 5821 TGTCTCTTTA TGAGGAGTTG TGGCCCGTTG TCAGGCAACG TGGCGTGGTG TGCACTGTGT 5881 TTGCTGACGC AACCCCCACT GGTTGGGGCA TTGCCACCAC CTGTCAGCTC CTTTCCGGGA 5941 CTTTCGCTTT CCCCCTCCCT ATTGCCACGG CGGAACTCAT CGCCGCCTGC CTTGCCCGCT 6001 GCTGGACAGG GGCTCGGCTG TTGGGCACTG ACAATTCCGT GGTGTTGTCG GGGAAGCTGA 6061 CGTCCTTTCC ATGGCTGCTC GCCTGTGTTG CCACCTGGAT TCTGCGCGGG ACGTCCTTCT 6121 GCTACGTCCC TTCGGCCCTC AATCCAGCGG ACCTTCCTTC CCGCGGCCTG CTGCCGGCTC 6181 TGCGGCCTCT TCCGCGTCTT CGAGATCTGC CTCGACTGTG CCTTCTAGTT GCCAGCCATC 6241 TGTTGTTTGC CCCTCCCCCG TGCCTTCCTT GACCCTGGAA GGTGCCACTC CCACTGTCCT 6301 TTCCTAATAA AATGAGGAAA TTGCATCGCA TTGTCTGAGT AGGTGTCATT CTATTCTGGG 6361 GGGTGGGGTG GGGCAGGACA GCAAGGGGGA GGATTGGGAA GACAATAGCA GGCATGCTGG 6421 GGACTCGAGT TAAGGGCGAA TTGCCGATTA GGATCTTCCT AGAGCATGGC TACGTAGATA 6481 AGTAGCATGG CGGGTTAATC ATTAACTACA AGGAACCCCT AGTGATGGAG TTGGCCACTC 6541 CCTCTCTGCG CGCTCGCTCG CTCACTGAGG CCGGGCGACC AAAGGTCGCC CGACGCCCGG 6601 GCTTTGCCCG GGCGGCCTCA GTGAGCGAGC GAGCGCGCAG CCTTAATTAA GAGCTCATGG 6661 CGCGCCTAGG CCTTGACGGC CTTCCGCCAA TTCGCCCTAT AGTGAGTCGT ATTACGTCGC 6721 GCTCACTGGC CGTCGTTTTA CAACGTCGTG ACTGGGAAAA CCCTGGCGTT ACCCAACTTA 6781 ATCGCCTTGC AGCACATCCC CCTTTCGCCA GCTGGCGTAA TAGCGAAGAG GCCCGCACCG 6841 AAACGCCCTT CCCAACAGTT GCGCAGCCTG AATGGCGAAT GGGAGCGCCC TGTAGCGGCC 6901 ACTCAACCCT ATCTCGGTCT ATTCTTTTGA TTTATAAGGG ATTTTGCCGA TTTCGGCCTA 6961 TTGGTTAAAA AATGAGCTGA TTTAACAAAA ATTTAACGCG AATTTTAACA AAATATTAAC 7021 GCTTACAATT TAG //