LOCUS NEU1-pGEc1-DEST 5740 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(5218..5347) /note="3'-ITR" misc_feature complement(4916..5130) /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 4368..4909 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_recomb 4273..4295 /label=attB2 /note="attB2 site" misc_feature 4305..4328 /note="8xHis Tag" misc_feature 4332..4355 /note="StrepTag" PCR_frame 3002..4273 /note="Reading Frame" /set="Set 1" source 3006..4250 /dnas_title="NEU1-pGEc1-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:1553 IMAGE:3506824" /tissue_type="Kidney, renal cell adenocarcinoma" /clone_lib="NIH_MGC_14" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 3006..4250 /dnas_title="NEU1" /gene="NEU1" /gene_synonym="SIAL1" /db_xref="GeneID:4758" /db_xref="HGNC:7758" /db_xref="MIM:608272" CDS 3006..4250 /dnas_title="sialidase 1 (lysosomal sialidase)" /gene="NEU1" /gene_synonym="SIAL1" /codon_start=1 /product="sialidase 1 (lysosomal sialidase)" /protein_id="AAH00722.1" /db_xref="GI:12653863" /db_xref="GeneID:4758" /db_xref="HGNC:7758" /db_xref="MIM:608272" /translation="MTGERPSTALPDRRWGPRILGFWGGCRVWVFAAIFLLLSLAASW SKAENDFGLVQPLVTMEQLLWVSGRQIGSVDTFRIPLITATPRGTLLAFAEARKMSSS DEGAKFIALRRSMDQGSTWSPTAFIVNDGDVPDGLNLGAVVSDVETGVVFLFYSLCAH KAGCQVASTMLVWSKDDGVSWSTPRNLSLDIGTEVFAPGPGSGIQKQREPRKGRLIVC GHGTLERDGVFCLLSDDHGASWRYGSGVSGIPYGQPKQENDFNPDECQPYELPDGSVV INARNQNNYHCHCRIVLRSYDACDTLRPRDVTFDPELVDPVVAAGAVVTSSGIVFFSN PAHPEFRVNLTLRWSFSNGTSWRKETVQLWPGPSGYSSLATLEGSMDGEEQAPQLYVL YEKGRNHYTESISVAKISVYGTL" misc_feature 3006..4250 /note="NEU1 coding region" misc_feature 4251..4271 /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..4280) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..4280) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(4281..5740,1..2991) /source="pGEc1-DEST" /type="Destination Vector" clone 2992..4280 /source="Neu1-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 TCGCCatgac tggggagcga cccagcacgg cgctcccgga cagacgctgg gggccgcgga 3061 ttctgggctt ctggggaggc tgtagggttt gggtgtttgc cgcgatcttc ctgctgctgt 3121 ctctggcagc ctcctggtcc aaggctgaga acgacttcgg tctggtgcag ccgctggtga 3181 ccatggagca actgctgtgg gtgagcggga gacagatcgg ctcagtggac accttccgca 3241 tcccgctcat cacagccact ccgcggggca ctcttctcgc ctttgctgag gcgaggaaaa 3301 tgtcctcatc cgatgagggg gccaagttca tcgccctgcg gaggtccatg gaccagggca 3361 gcacatggtc tcctacagcg ttcattgtca atgatgggga tgtccccgat gggctgaacc 3421 ttggggcagt agtgagcgat gttgagacag gagtagtatt tcttttctac tccctttgtg 3481 ctcacaaggc cggctgccag gtggcctcta ccatgttggt atggagcaag gatgatggtg 3541 tttcctggag cacaccccgg aatctctccc tggatattgg cactgaagtg tttgcccctg 3601 gaccgggctc tggtattcag aaacagcggg agccacggaa gggccgcctc atcgtgtgtg 3661 gccatgggac gctggagcgg gacggagtct tctgtctcct cagcgatgat catggtgcct 3721 cctggcgcta cggaagtggg gtcagcggca tcccctacgg tcagcccaag caggaaaatg 3781 atttcaatcc tgatgaatgc cagccctatg agctcccaga tggctcagtc gtcatcaatg 3841 cccgaaacca gaacaactac cactgccact gccgaattgt cctccgcagc tatgatgcct 3901 gtgatacact aaggccccgt gatgtgacct tcgaccctga gctcgtggac cctgtggtag 3961 ctgcaggagc tgtagtcacc agctccggca ttgtcttctt ctccaaccca gcacatccag 4021 agttccgagt gaacctgacc ctgcgatgga gcttcagcaa tggtacctca tggcggaaag 4081 agacagtcca gctatggcca ggccccagtg gctattcatc cctggcaacc ctggagggca 4141 gcatggatgg agaggagcag gccccccagc tctacgtcct gtatgagaaa ggccggaacc 4201 actacacaga gagcatctcc gtggccaaaa tcagtgtcta tgggacactc gaaaacttgt 4261 actttcaagg ctCCAGCTTT CTTGTACAAA GTGGTGATGG CCGCCATCAT CATCATCATC 4321 ATCATCATGC GTGGAGCCAC CCGCAGTTCG AAAAATAATA AGGATCCAAT CAACCTCTGG 4381 ATTACAAAAT TTGTGAAAGA TTGACTGGTA TTCTTAACTA TGTTGCTCCT TTTACGCTAT 4441 GTGGATACGC TGCTTTAATG CCTTTGTATC ATGCTATTGC TTCCCGTATG GCTTTCATTT 4501 TCTCCTCCTT GTATAAATCC TGGTTGCTGT CTCTTTATGA GGAGTTGTGG CCCGTTGTCA 4561 GGCAACGTGG CGTGGTGTGC ACTGTGTTTG CTGACGCAAC CCCCACTGGT TGGGGCATTG 4621 CCACCACCTG TCAGCTCCTT TCCGGGACTT TCGCTTTCCC CCTCCCTATT GCCACGGCGG 4681 AACTCATCGC CGCCTGCCTT GCCCGCTGCT GGACAGGGGC TCGGCTGTTG GGCACTGACA 4741 ATTCCGTGGT GTTGTCGGGG AAGCTGACGT CCTTTCCATG GCTGCTCGCC TGTGTTGCCA 4801 CCTGGATTCT GCGCGGGACG TCCTTCTGCT ACGTCCCTTC GGCCCTCAAT CCAGCGGACC 4861 TTCCTTCCCG CGGCCTGCTG CCGGCTCTGC GGCCTCTTCC GCGTCTTCGA GATCTGCCTC 4921 GACTGTGCCT TCTAGTTGCC AGCCATCTGT TGTTTGCCCC TCCCCCGTGC CTTCCTTGAC 4981 CCTGGAAGGT GCCACTCCCA CTGTCCTTTC CTAATAAAAT GAGGAAATTG CATCGCATTG 5041 TCTGAGTAGG TGTCATTCTA TTCTGGGGGG TGGGGTGGGG CAGGACAGCA AGGGGGAGGA 5101 TTGGGAAGAC AATAGCAGGC ATGCTGGGGA CTCGAGTTAA GGGCGAATTG CCGATTAGGA 5161 TCTTCCTAGA GCATGGCTAC GTAGATAAGT AGCATGGCGG GTTAATCATT AACTACAAGG 5221 AACCCCTAGT GATGGAGTTG GCCACTCCCT CTCTGCGCGC TCGCTCGCTC ACTGAGGCCG 5281 GGCGACCAAA GGTCGCCCGA CGCCCGGGCT TTGCCCGGGC GGCCTCAGTG AGCGAGCGAG 5341 CGCGCAGCCT TAATTAAGAG CTCATGGCGC GCCTAGGCCT TGACGGCCTT CCGCCAATTC 5401 GCCCTATAGT GAGTCGTATT ACGTCGCGCT CACTGGCCGT CGTTTTACAA CGTCGTGACT 5461 GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT 5521 GGCGTAATAG CGAAGAGGCC CGCACCGAAA CGCCCTTCCC AACAGTTGCG CAGCCTGAAT 5581 GGCGAATGGG AGCGCCCTGT AGCGGCCACT CAACCCTATC TCGGTCTATT CTTTTGATTT 5641 ATAAGGGATT TTGCCGATTT CGGCCTATTG GTTAAAAAAT GAGCTGATTT AACAAAAATT 5701 TAACGCGAAT TTTAACAAAA TATTAACGCT TACAATTTAG //