LOCUS NEU1-pGEc2-DEST 6466 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(5944..6073) /note="3'-ITR" misc_feature complement(5642..5856) /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 5094..5635 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 4273..4295 /note="attB2" misc_feature 5073..5078 /note="BstB1 site" misc_feature 4296..4991 /note="Super GFP" misc_feature 4992..5042 /note="AviTag" misc_feature 5043..5066 /note="His Tag" PCR_frame 3002..4273 /note="Reading Frame" /set="Set 1" source 3006..4250 /dnas_title="NEU1-pGEc2-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..6466,1..2991) /source="pGEc2-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 GTGGTATGAG CAAGGGCGAG GAACTGTTCA 4321 CCGGCGTGGT GCCCATCCTG GTGGAACTGG ATGGCGACGT GAACGGCCAC AAGTTCTCTG 4381 TGCGGGGAGA GGGCGAAGGC GACGCCACAA ATGGCAAGCT GACCCTGAAG TTCATCTGCA 4441 CCACCGGCAA GCTGCCCGTG CCTTGGCCTA CCCTCGTGAC CACACTGACC TACGGCGTGC 4501 AGTGCTTCAG CAGATACCCC GACCATATGA AGCGGCACGA CTTCTTCAAG AGCGCCATGC 4561 CCGAGGGCTA CGTGCAGGAA CGGACCATCA GCTTCAAGGA CGACGGCACC TACAAGACCA 4621 GAGCCGAAGT GAAGTTCGAG GGCGACACCC TCGTGAACCG GATCGAGCTG AAGGGCATCG 4681 ATTTCAAAGA GGACGGCAAC ATCCTGGGCC ACAAGCTGGA GTACAACTTC AACAGCCACA 4741 ACGTGTACAT CACCGCCGAC AAGCAGAAGA ACGGCATCAA GGCCAACTTC AAGATCCGGC 4801 ACAACGTGGA AGATGGCAGC GTGCAGCTGG CCGACCACTA CCAGCAGAAC ACCCCCATCG 4861 GAGATGGCCC CGTGCTGCTG CCCGACAACC ACTACCTGAG CACCCAGAGC GTGCTGAGCA 4921 AGGACCCCAA CGAGAAGCGG GACCACATGG TGCTGCTGGA ATTCGTGACC GCCGCTGGCA 4981 TCACCCACGG CATGAGCGGC CTGAACGACA TCTTCGAGGC CCAGAAAATC GAGTGGCACG 5041 AGCACCACCA TCATCACCAT CACCACTGAT GATTCGAAAA ATAATAAGGA TCCAATCAAC 5101 CTCTGGATTA CAAAATTTGT GAAAGATTGA CTGGTATTCT TAACTATGTT GCTCCTTTTA 5161 CGCTATGTGG ATACGCTGCT TTAATGCCTT TGTATCATGC TATTGCTTCC CGTATGGCTT 5221 TCATTTTCTC CTCCTTGTAT AAATCCTGGT TGCTGTCTCT TTATGAGGAG TTGTGGCCCG 5281 TTGTCAGGCA ACGTGGCGTG GTGTGCACTG TGTTTGCTGA CGCAACCCCC ACTGGTTGGG 5341 GCATTGCCAC CACCTGTCAG CTCCTTTCCG GGACTTTCGC TTTCCCCCTC CCTATTGCCA 5401 CGGCGGAACT CATCGCCGCC TGCCTTGCCC GCTGCTGGAC AGGGGCTCGG CTGTTGGGCA 5461 CTGACAATTC CGTGGTGTTG TCGGGGAAGC TGACGTCCTT TCCATGGCTG CTCGCCTGTG 5521 TTGCCACCTG GATTCTGCGC GGGACGTCCT TCTGCTACGT CCCTTCGGCC CTCAATCCAG 5581 CGGACCTTCC TTCCCGCGGC CTGCTGCCGG CTCTGCGGCC TCTTCCGCGT CTTCGAGATC 5641 TGCCTCGACT GTGCCTTCTA GTTGCCAGCC ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC 5701 CTTGACCCTG GAAGGTGCCA CTCCCACTGT CCTTTCCTAA TAAAATGAGG AAATTGCATC 5761 GCATTGTCTG AGTAGGTGTC ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG 5821 GGAGGATTGG GAAGACAATA GCAGGCATGC TGGGGACTCG AGTTAAGGGC GAATTGCCGA 5881 TTAGGATCTT CCTAGAGCAT GGCTACGTAG ATAAGTAGCA TGGCGGGTTA ATCATTAACT 5941 ACAAGGAACC CCTAGTGATG GAGTTGGCCA CTCCCTCTCT GCGCGCTCGC TCGCTCACTG 6001 AGGCCGGGCG ACCAAAGGTC GCCCGACGCC CGGGCTTTGC CCGGGCGGCC TCAGTGAGCG 6061 AGCGAGCGCG CAGCCTTAAT TAAGAGCTCA TGGCGCGCCT AGGCCTTGAC GGCCTTCCGC 6121 CAATTCGCCC TATAGTGAGT CGTATTACGT CGCGCTCACT GGCCGTCGTT TTACAACGTC 6181 GTGACTGGGA AAACCCTGGC GTTACCCAAC TTAATCGCCT TGCAGCACAT CCCCCTTTCG 6241 CCAGCTGGCG TAATAGCGAA GAGGCCCGCA CCGAAACGCC CTTCCCAACA GTTGCGCAGC 6301 CTGAATGGCG AATGGGAGCG CCCTGTAGCG GCCACTCAAC CCTATCTCGG TCTATTCTTT 6361 TGATTTATAA GGGATTTTGC CGATTTCGGC CTATTGGTTA AAAAATGAGC TGATTTAACA 6421 AAAATTTAAC GCGAATTTTA ACAAAATATT AACGCTTACA ATTTAG //