LOCUS Exported 9830 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses full-length Nephrin precursor ectodomain in mammalian cells. C-terminal rat Cd4d3+4 tag, biotinylation sequence and His tag.. ACCESSION . VERSION . KEYWORDS NPHS-bio-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9830) AUTHORS Sun Y, Vandenbriele C, Kauskot A, Verhamme P, Hoylaerts MF, Wright GJ TITLE A human platelet receptor protein microarray identifies FcepsilonR1alpha as an activating PEAR1 ligand. JOURNAL Mol Cell Proteomics. 2015 Feb 23. pii: mcp.M114.046946. PUBMED 25713122 REFERENCE 2 (bases 1 to 9830) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9830 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 644..661 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 713..768 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(767..786) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3004..3022) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3090..3194 /gene="bla" /label=AmpR promoter CDS 3195..4055 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3413..3432) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4226..4814 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4715..4734 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4968..4985 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5102..5123 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5138..5168 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5176..5192 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 5181..5203 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5200..5216 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5200..5216 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 5487..5866 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5867..6070 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6021..6041 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 6067..6091 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 6572..6591 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 6625..6634 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 gtcgacctcc atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact 61 caaccttgga gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc 121 ttcccagtcc tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac 181 tagcaacccc aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt 241 ctcccttcag tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta 301 tcaggaagtg aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg 361 tgaggtgatg ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga 421 ggccagggtc tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg 481 gcaatgtcta ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc 541 caaagggttg aattccggca gcctgcacca catcctggac gcccagaaga tgctgtggaa 601 ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc atcaccacca 661 ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg ctaataaagg 721 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca 781 tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag aggatcgatc 841 cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg cggggcagtg 901 catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc cacatgtgac 961 acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct tataaatgga 1021 tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc agtctgtgga 1081 ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca ccaatgctgg 1141 gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa cgtcaatcag 1201 aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa tagtgtttat 1261 aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta gtatatacta 1321 tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat gctatcgaat 1381 tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga gctgtcacgg 1441 ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt cacaagggca 1501 gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt 1561 ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg tatgtgaggt 1621 gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg gggttcagtg 1681 gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa taaatactag 1741 tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt ggggacaagc 1801 cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac ataatcctag 1861 tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga caggtgaacc 1921 atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca gcagcggact 1981 ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa tggggcccat 2041 aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca cgcgtcagcc 2101 cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact tggctgattg 2161 taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg 2221 ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct gcgatctgga 2281 ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt cctcatattc 2341 acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata ctacccaaat 2401 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat 2461 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat 2521 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat 2581 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctaatagag 2641 attagggtag tatatgctat cctaatttat atctgggtag catatactac ccaaatatct 2701 ggatagcata tgctatccta atctatatct gggtagcata tgctatccta atctatatct 2761 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct 2821 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct 2881 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc 2941 gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa 3001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 3061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 3121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 3181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 3241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 3301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 3361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 3421 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt 3481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 3541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 3601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 3661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 3721 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac 3781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 3841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 3901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 3961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 4021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 4081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 4141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 4201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 4261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 4321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 4381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 4441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 4561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 4801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 4861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 4921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 4981 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 5041 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 5101 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 5161 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 5221 attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca 5281 gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta 5341 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa 5401 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg 5461 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc 5521 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 5581 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 5641 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 5701 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga 5761 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt 5821 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 5881 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 5941 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 6001 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 6061 aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg ctgtctgcga 6121 gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg 6181 gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg 6241 catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta 6301 ggctgagcac cgtggcgggc ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc 6361 tgctgatgat gtaattaaag taggcggtct tgagacggcg gatggtcgag gtgaggtgtg 6421 gcaggcttga gatccagctg ttggggtgag tactccctct caaaagcggg cattacttct 6481 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgatctg 6541 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc 6601 caggtccaag tttaaactgc ggccgccacc atggccctgg gcaccacact gagagccagc 6661 ctgctgctgc tgggcctgct gacagaagga ctggcacagc tggctatccc tgccagcgtg 6721 cctagaggct tctgggccct gcctgagaac ctgacagtgg tggagggcgc cagcgtggag 6781 ctgagatgcg gcgtgagcac acctggatct gccgtgcagt gggccaagga tggcctgctg 6841 ctcggccccg atcccagaat ccccggcttc ccccggtaca gactggaagg cgaccccgcc 6901 agaggcgagt tccatctgca catcgaggcc tgcgacctgt ctgacgacgc cgagtacgag 6961 tgccaggtcg gcagatccga gatgggcccc gagctggtgt cccccagagt gatcctgagc 7021 atcctggtgc cccccaagct gctgctcctg acccctgagg ccggcaccat ggtcacatgg 7081 gtcgcaggcc aggaatacgt cgtcaactgc gtgagcggcg acgccaagcc tgcccccgac 7141 atcaccatcc tgctgtccgg ccagaccatc agcgacatca gcgccaacgt gaacgagggc 7201 agccagcaga aactgttcac cgtggaggcc accgccagag tgacccccag aagcagcgac 7261 aaccggcagc tgctcgtgtg tgaagccagc agccctgccc tggaagcccc catcaaggcc 7321 agcttcaccg tgaacgtgct gttcccccct ggccctcctg tgatcgagtg gcctggcctg 7381 gatgaaggac acgtgcgggc tggccagagc ctggaactgc cttgtgtggc taggggaggt 7441 aatcctctcg ccaccctgca gtggctgaag aacggccagc ctgtgtccac agcctggggc 7501 acagagcata cccaggccgt ggccagatcc gtgctggtca tgaccgtgcg gcctgaggat 7561 catggcgccc agctgtcctg cgaggcccac aatagcgtgt ccgccggcac ccaggaacac 7621 ggcatcaccc tccaggtcac cttccccccc agcgccatca tcatcctggg cagcgccagc 7681 cagaccgaga acaagaacgt gaccctgagc tgcgtgagca agtcatctcg gcccagagtg 7741 ctgctgagat ggtggctggg gtggagacag ctgctgccca tggaagagac agtgatggac 7801 ggcctgcacg gcggccacat cagcatgagc aacctgacct ttctggccag acgggaggac 7861 aatggcctga ccctgacatg cgaggccttc tccgaggcct tcaccaaaga gacattcaag 7921 aagtccctga tcctgaacgt gaagtacccc gcccagaagc tgtggatcga gggcccacct 7981 gagggccaga agctgagagc cggcacaaga gtgcggctcg tgtgtctggc catcggcggc 8041 aaccctgagc ccagcctgat gtggtacaag gacagccgga ccgtgaccga gagcagactg 8101 ccccaggaaa gcagaagagt ccacctgggc tccgtggaga agtccggcag caccttcagc 8161 cgggagctgg tgctggtcac cggccccagc gataaccagg ccaagttcac atgcaaggcc 8221 ggacagctct ctgcctctac acagctggcc gtgcagttcc ctcccaccaa cgtgaccatc 8281 ctggccaacg cctctgctct gaggcctggc gacgccctga acctgacctg tgtgtccgtg 8341 agcagcaacc cccccgtgaa cctgtcctgg gacaaagagg gcgagaggct ggaaggtgtc 8401 gccgctcctc ccagaagggc cccctttaag ggatctgccg ctgccagatc agtcctgctc 8461 caggtgtcca gcagggacca cggccagaga gtgacctgta gagcccacag cgccgagctg 8521 agagaaaccg tgtccagctt ctacagactg aatgtgctgt acaggcctga gttcctggga 8581 gaacaggtgc tggtcgtgac agccgtggag cagggcgaag ctctgctgcc cgtgtccgtg 8641 tctgccaatc ccgccccaga ggccttcaac tggaccttcc ggggctacag actgtctcct 8701 gccggcggac ccagacacag aatcctgagc agcggagccc tgcacctgtg gaacgtgacc 8761 agagccgacg acggcctgta ccagctgcac tgccagaata gcgagggcac cgctgaagcc 8821 agactgagac tggacgtgca ctacgccccc accatcagag ccctgcagga ccccaccgaa 8881 gtgaatgtgg gcggctctgt ggacatcgtg tgcaccgtgg acgccaaccc catcctgccc 8941 ggcatgttca actgggagcg gctgggcgag gacgaggaag atcagtccct ggacgacatg 9001 gaaaagatca gcagaggccc caccggcaga ctgagaatcc accacgccaa gctggctcag 9061 gctggcgcct accagtgcat cgtggacaac ggcgtggccc ctcctgctcg gagactgctg 9121 cggctggtcg tgcgattcgc acctcaggtg gagcacccca cccctctgac aaaagtggcc 9181 gctgccggcg atagcacaag cagcgccaca ctgcactgcc gcgctagagg cgtgcccaac 9241 atcgtgttta cctggaccaa gaacggcgtg ccactcgatc tgcaggatcc ccggtacacc 9301 gagcacacat accaccaggg cggcgtgcac agctctctgc tgacaatcgc caacgtgtct 9361 gccgcccagg actacgccct gttcacctgt accgccacaa acgccctggg ctctgaccag 9421 accaacatcc agctggtgtc tatcagcaga cccgaccccc ctagcggcct gaaggtggtg 9481 tccctgaccc ctcacagcgt gggcctggaa tggaagcctg gcttcgacgg cggcctgccc 9541 cagcggttct gcatcagata cgaggccctg gggacccctg gcttccacta cgtggacgtg 9601 gtgcctcccc aggccaccac ctttaccctg accggcctgc agcccagcac cagatacaga 9661 gtgtggctgc tcgccagcaa tgccctgggc gatagcggcc tggccgataa gggcacccag 9721 ctgcccatca ccacacctgg cctgcaccag cctagcggcg agccagagga ccagctgcct 9781 accgagcctc ctagcggccc ttctggcctg cctctgctgc ctggcgcgcc //