LOCUS Exported 7697 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION To express human Nup93 in mammalian cells. ACCESSION . VERSION . KEYWORDS pEGFP-C1X-Nup93 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7697) AUTHORS Tie HC, Madugula V, Lu L TITLE The development of a single molecule fluorescence standard and its application in estimating the stoichiometry of the nuclear pore complex. JOURNAL Biochem Biophys Res Commun. 2016 Sep 30;478(4):1694-9. doi: 10.1016/j.bbrc.2016.09.005. Epub 2016 Sep 6. PUBMED 27613095 REFERENCE 2 (bases 1 to 7697) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7697 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 484..605 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(521..540) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 575..594 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(612..1067) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(749..770) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 961..980 /label=F1ori-R /note="F1 origin, reverse primer" promoter 1094..1198 /gene="bla" /label=AmpR promoter promoter 1200..1557 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1223..1243) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1408..1543 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1470..1489 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 1592..2386 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1646..1665) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2256..2275 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2574..2593) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2618..2665 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2994..3582 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3483..3502 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 3757..4060 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4061..4264 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4215..4235 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 4303..4312 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4309..5025 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4309..5025 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4354..4375) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4615..4634) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4962..4983 /label=EGFP-C /note="EGFP, forward primer" ORIGIN 1 tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac atcctgggca 61 ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac aaggccaaga 121 atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc gtgcagctgg 181 ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg cccgataacc 241 actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc gatcacatga 301 tctacttcgg cttcgtgacc tctagagtat ccctcgagga tccgagctcc gcggagatcc 361 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct 421 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 481 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 601 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 661 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 721 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 781 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 841 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 901 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 961 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 1021 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt 1081 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 1141 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc 1201 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 1261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 1321 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 1381 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 1441 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 1501 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat 1561 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 1621 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 1681 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 1741 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct 1801 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 1861 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 1921 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 1981 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 2041 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 2101 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 2161 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 2221 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 2281 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 2341 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 2401 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 2461 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 2521 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca 2581 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 2641 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 2701 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 2761 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 2821 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa 2881 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 2941 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 3001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 3061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 3121 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 3181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 3241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 3301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 3361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 3421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 3481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 3541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 3601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 3661 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac 3721 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg 3781 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 3841 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 3901 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 3961 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 4021 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 4081 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 4141 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa 4201 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 4261 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag 4321 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 4381 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 4441 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 4501 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 4561 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 4621 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 4681 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 4741 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg 4801 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 4861 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 4921 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 4981 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct 5041 cgacttcccg ggatgacggc tgtagcacaa ggatctgcat ctccaatgga tactgagggg 5101 tttggtgagc tccttcagca agctgaacag cttgctgctg agactgaggg catctcagag 5161 cttccccatg tggaacggaa cttacaggag atccagcagg cgggagagcg cctgcgttcc 5221 cgtaccctaa cacgcacgtc ccaggagacg gcagatgtca aggcgtcagt tctcctcggg 5281 tctcggggac ttgacatatc ccacatctcc cagcgattgg agagtctgag tgcagccacc 5341 acctttgagc ctcttgagcc tgtgaaggac actgacattc agggcttcct gaagaatgag 5401 aaggacaatg ccctgctgtc tgccatcgaa gagtcccgga agaggacctt cggcatggct 5461 gaggagtacc atcgggagtc aatgttggtt gagtgggagc aagtgaaaca gcgaattctg 5521 cacacactgc tggcatcagg agaagacgcc cttgacttta ctcaagaaag cgagccaagc 5581 tacatcagtg atgtgggacc ccctggtcga agctctctgg ataacatcga gatggcctat 5641 gcgcggcaaa tttatatcta taatgagaaa attgtaaatg gacacctgca gcctaacctg 5701 gtggaccttt gtgcttccgt cgcagagctc gatgataaga gcatttccga catgtggacc 5761 atggtaaaac aaatgacaga cgtgttgttg acaccggcaa cggatgccct gaagaaccgc 5821 agcagcgtgg aagtgcgcat ggagtttgtc aggcaggcct tggcgtacct tgagcagagt 5881 tataagaatt acacccttgt gactgtcttt ggaaatttgc atcaggccca gctgggcggg 5941 gtgcctggga cttaccaatt ggttcgaagt ttcctgaaca ttaaactgcc agctcccttg 6001 cctggactac aggatggaga ggtggaaggc catcctgtgt gggcgctaat ttactactgc 6061 atgcgctgtg gagacctgct tgccgcttca caggtagtta atcgagccca gcaccagctg 6121 ggagagttta aaacctggtt ccaggagtac atgaacagca aggacagaag attgtcccca 6181 gctacggaaa acaagctccg gctgcattac cgtagggccc tcaggaacaa tacagatccc 6241 tacaagcggg ccgtgtactg tatcattggc agatgtgacg tcaccgacaa ccagagtgaa 6301 gtggcggaca aaactgagga ttacctgtgg ctgaagttga accaagtgtg ttttgacgac 6361 gatggcacca gctccccaca agacaggctc actctctcac agttccagaa gcagttgttg 6421 gaagactatg gcgagtccca ctttacggtg aaccagcaac ccttcctcta cttccaagtc 6481 ctgttcctga cagcgcagtt tgaagcagca gttgcctttc ttttccgcat ggagcggctg 6541 cgctgccatg ctgtccatgt agcactggtg ctgtttgagc tgaagctgct tttaaagtcc 6601 tctggacaga gggctcagct cctcagccac gagcctggtg accctccttg cttgcggcgg 6661 ctgaacttcg tgcggctcct catgctgtac acccggaagt ttgagtccac ggacccaagg 6721 gaggccctcc agtacttcta tttcctcagg gatgagaaag atagtcaagg agaaaacatg 6781 tttctgcgct gtgtgagtga gcttgtgatt gaaagccgag agttcgatat gattcttggg 6841 aaactagaga atgacggaag tagaaagcct ggagtcatag ataagtttac tagtgacaca 6901 aagcctatta tcaacaaagt tgcttctgtg gcagaaaata aaggactgtt tgaagaggca 6961 gcaaagctgt atgaccttgc caagaatgct gacaaggtac tggagctgat gaacaaactg 7021 ctgagccctg tcgtccccca gatcagtgcc ccgcaatcca acaaggagag gctgaagaac 7081 atggcactct ccattgccga acggtatagg gctcaaggaa taagcgcaaa taaatttgtg 7141 gactccacgt tctatcttct tttggacttg atcacctttt ttgacgagta tcatagtggt 7201 catattgata gagcttttga tatcattgag cgcttgaagc tggtgcccct gaatcaggaa 7261 agtgtggaag agagagtggc tgccttcaga aatttcagtg atgaaatcag gcacaacctc 7321 tcagaagtgc ttcttgccac catgaacatc ttgttcacac agtttaagag gctcaagggg 7381 acaagtccat cctcgtcatc caggccccag cgagtcatcg aggaccgcga ctctcaactc 7441 cgaagtcaag cccgcactct gattaccttt gctggaatga taccataccg aacgtctggg 7501 gacaccaatg cgaggctggt gcagatggag gtcctcatga attaagtgcc atgctttgtg 7561 ggagtctggg tcggcacact gtcagtacat caggcacatg ggcccactag gctggggttt 7621 ctggttttgt ttctgttgtg ttttgttttg gtttctgtat tatgtatttt tgtcaacgcc 7681 aataaatttc tttgatt //