LOCUS NEU4-pGEc2-DEST 6709 bp DNA circular 24-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6187..6316) /note="3'-ITR" misc_feature complement(5885..6099) /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 5337..5878 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 4516..4538 /note="attB2" misc_feature 5316..5321 /note="BstB1 site" misc_feature 4539..5234 /note="Super GFP" misc_feature 5235..5285 /note="AviTag" misc_feature 5286..5309 /note="His Tag" PCR_frame 3002..4516 /note="Reading Frame" /set="Set 1" source 3006..4493 /dnas_title="NEU4-pGEc2-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:18222 IMAGE:4156395" /tissue_type="Brain, anaplastic oligodendroglioma with 1p/19q loss" /clone_lib="NCI_CGAP_Brn67" /lab_host="DH10B" /note="Vector: pCMV-SPORT6" gene 3006..4493 /dnas_title="NEU4" /gene="NEU4" /gene_synonym="MGC18222" /db_xref="GeneID:129807" /db_xref="HGNC:21328" /db_xref="MIM:608527" CDS 3006..4493 /dnas_title="sialidase 4" /gene="NEU4" /gene_synonym="MGC18222" /codon_start=1 /product="sialidase 4" /protein_id="AAH12899.2" /db_xref="GI:49208223" /db_xref="GeneID:129807" /db_xref="HGNC:21328" /db_xref="MIM:608527" /translation="MMSSAAFPRWLSMGVPRTPSRTVLFERERTGLTYRVPSLLPVPP GPTLLAFVEQRLSPDDSHAHRLVLRRGTLAGGSVRWGALHVLGTAALAEHRSMNPCPV HDAGTGTVFLFFIAVLGHTPEAVQIATGRNAARLCCVASRDAGLSWGSARDLTEEAIG GAVQDWATFAVGPGHGVQLPSGRLLVPAYTYRVDRRECFGKICRTSPHSFAFYSDDHG RTWRCGGLVPNLRSGECQLAAVDGGQAGSFLYCNARSPLGSRVQALSTDEGTSFLPAE RVASLPETAWGCQGSIVGFPAPAPNRPRDDSWSVGPGSPLQPPLLGPGVHEPPEEAAV DPRGGQVPGGPFSRLQPRGDGPRQPGPRPGVSGDVGSWTLALPMPFAAPPQSPTWLLY SHPVGRRARLHMGIRLSQSPLDPRSWTEPWVIYEGPSGYSDLASIGPAPEGGLVFACL YESGARTSYDEISFCTFSLREVLENVPASPKPPNLGDKPRGCCWPS" misc_feature 3006..4493 /note="Neu4 coding region" misc_feature 4494..4514 /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..4523) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..4523) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(4524..6709,1..2991) /source="pGEc2-DEST" /type="Destination Vector" clone 2992..4523 /source="Neu4 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 TCGCCatgat gagctctgca gccttcccaa ggtggctgag catgggggtc cctcgtaccc 3061 cttcacggac agtgctcttc gagcgggaga ggacgggcct gacctaccgc gtgccctcgc 3121 tgctccccgt gccccccggg cccaccctgc tggcctttgt ggagcagcgg ctcagccctg 3181 acgactccca cgcccaccgc ctggtgctga ggaggggcac gctggccggg ggctccgtgc 3241 ggtggggtgc cctgcacgtg ctggggacag cagccctggc ggagcaccgg tccatgaacc 3301 cctgccctgt gcacgatgct ggcacgggca ccgtcttcct cttcttcatc gcggtgctgg 3361 gccacacgcc tgaggccgtg cagatcgcca cgggaaggaa cgccgcgcgc ctctgctgtg 3421 tggccagccg tgacgccggc ctctcgtggg gcagcgcccg ggacctcacc gaggaggcca 3481 tcggtggtgc cgtgcaggac tgggccacat tcgctgtggg tcccggccac ggcgtgcagc 3541 tgccctcagg ccgcctgctg gtacccgcct acacctaccg cgtggaccgc cgagagtgtt 3601 ttggcaagat ctgccggacc agccctcact ccttcgcctt ctacagcgat gaccacggcc 3661 gcacctggcg ctgtggaggc ctcgtgccca acctgcgctc aggcgagtgc cagctggcgg 3721 cggtggacgg tgggcaggcc ggcagcttcc tctactgcaa tgcccggagc ccactgggca 3781 gccgtgtgca ggcgctcagc actgacgagg gcacctcctt cctgcccgca gagcgcgtgg 3841 cttccctgcc cgagactgcc tggggctgcc agggcagcat cgtgggcttc ccagcccccg 3901 cccccaacag gccacgggat gacagttggt cagtgggccc cgggagtccc ctccagcctc 3961 cactcctcgg tcctggagtc cacgaacccc cagaggaggc tgctgtagac ccccgtggag 4021 gccaggtgcc tggtgggccc ttcagccgtc tgcagcctcg gggggatggc cccaggcagc 4081 ctggccccag gcctggggtc agtggggatg tggggtcctg gaccctggca ctccccatgc 4141 cctttgctgc cccgccccag agccccacgt ggctgctgta ctcccaccca gtggggcgca 4201 gggctcggct acacatgggt atccgcctga gccagtcccc gctggacccg cgcagctgga 4261 cagagccctg ggtgatctac gagggcccca gcggctactc cgacctggcg tccatcgggc 4321 cggctcctga ggggggcctg gtttttgcct gcctgtacga gagcggggcc aggacctcct 4381 atgatgagat ttccttttgt acattctccc tgcgtgaggt cctggagaac gtgcccgcca 4441 gccccaagcc gcccaacctt ggggacaagc ctcgggggtg ctgctggccc tccgaaaact 4501 tgtactttca aggctCCAGC TTTCTTGTAC AAAGTGGTAT GAGCAAGGGC GAGGAACTGT 4561 TCACCGGCGT GGTGCCCATC CTGGTGGAAC TGGATGGCGA CGTGAACGGC CACAAGTTCT 4621 CTGTGCGGGG AGAGGGCGAA GGCGACGCCA CAAATGGCAA GCTGACCCTG AAGTTCATCT 4681 GCACCACCGG CAAGCTGCCC GTGCCTTGGC CTACCCTCGT GACCACACTG ACCTACGGCG 4741 TGCAGTGCTT CAGCAGATAC CCCGACCATA TGAAGCGGCA CGACTTCTTC AAGAGCGCCA 4801 TGCCCGAGGG CTACGTGCAG GAACGGACCA TCAGCTTCAA GGACGACGGC ACCTACAAGA 4861 CCAGAGCCGA AGTGAAGTTC GAGGGCGACA CCCTCGTGAA CCGGATCGAG CTGAAGGGCA 4921 TCGATTTCAA AGAGGACGGC AACATCCTGG GCCACAAGCT GGAGTACAAC TTCAACAGCC 4981 ACAACGTGTA CATCACCGCC GACAAGCAGA AGAACGGCAT CAAGGCCAAC TTCAAGATCC 5041 GGCACAACGT GGAAGATGGC AGCGTGCAGC TGGCCGACCA CTACCAGCAG AACACCCCCA 5101 TCGGAGATGG CCCCGTGCTG CTGCCCGACA ACCACTACCT GAGCACCCAG AGCGTGCTGA 5161 GCAAGGACCC CAACGAGAAG CGGGACCACA TGGTGCTGCT GGAATTCGTG ACCGCCGCTG 5221 GCATCACCCA CGGCATGAGC GGCCTGAACG ACATCTTCGA GGCCCAGAAA ATCGAGTGGC 5281 ACGAGCACCA CCATCATCAC CATCACCACT GATGATTCGA AAAATAATAA GGATCCAATC 5341 AACCTCTGGA TTACAAAATT TGTGAAAGAT TGACTGGTAT TCTTAACTAT GTTGCTCCTT 5401 TTACGCTATG TGGATACGCT GCTTTAATGC CTTTGTATCA TGCTATTGCT TCCCGTATGG 5461 CTTTCATTTT CTCCTCCTTG TATAAATCCT GGTTGCTGTC TCTTTATGAG GAGTTGTGGC 5521 CCGTTGTCAG GCAACGTGGC GTGGTGTGCA CTGTGTTTGC TGACGCAACC CCCACTGGTT 5581 GGGGCATTGC CACCACCTGT CAGCTCCTTT CCGGGACTTT CGCTTTCCCC CTCCCTATTG 5641 CCACGGCGGA ACTCATCGCC GCCTGCCTTG CCCGCTGCTG GACAGGGGCT CGGCTGTTGG 5701 GCACTGACAA TTCCGTGGTG TTGTCGGGGA AGCTGACGTC CTTTCCATGG CTGCTCGCCT 5761 GTGTTGCCAC CTGGATTCTG CGCGGGACGT CCTTCTGCTA CGTCCCTTCG GCCCTCAATC 5821 CAGCGGACCT TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG GCCTCTTCCG CGTCTTCGAG 5881 ATCTGCCTCG ACTGTGCCTT CTAGTTGCCA GCCATCTGTT GTTTGCCCCT CCCCCGTGCC 5941 TTCCTTGACC CTGGAAGGTG CCACTCCCAC TGTCCTTTCC TAATAAAATG AGGAAATTGC 6001 ATCGCATTGT CTGAGTAGGT GTCATTCTAT TCTGGGGGGT GGGGTGGGGC AGGACAGCAA 6061 GGGGGAGGAT TGGGAAGACA ATAGCAGGCA TGCTGGGGAC TCGAGTTAAG GGCGAATTGC 6121 CGATTAGGAT CTTCCTAGAG CATGGCTACG TAGATAAGTA GCATGGCGGG TTAATCATTA 6181 ACTACAAGGA ACCCCTAGTG ATGGAGTTGG CCACTCCCTC TCTGCGCGCT CGCTCGCTCA 6241 CTGAGGCCGG GCGACCAAAG GTCGCCCGAC GCCCGGGCTT TGCCCGGGCG GCCTCAGTGA 6301 GCGAGCGAGC GCGCAGCCTT AATTAAGAGC TCATGGCGCG CCTAGGCCTT GACGGCCTTC 6361 CGCCAATTCG CCCTATAGTG AGTCGTATTA CGTCGCGCTC ACTGGCCGTC GTTTTACAAC 6421 GTCGTGACTG GGAAAACCCT GGCGTTACCC AACTTAATCG CCTTGCAGCA CATCCCCCTT 6481 TCGCCAGCTG GCGTAATAGC GAAGAGGCCC GCACCGAAAC GCCCTTCCCA ACAGTTGCGC 6541 AGCCTGAATG GCGAATGGGA GCGCCCTGTA GCGGCCACTC AACCCTATCT CGGTCTATTC 6601 TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG AGCTGATTTA 6661 ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTAG //