LOCUS       sequence-177620-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express human Nup35 in mammalian cells. It contains synonymous 
            mutations that are not targeted by shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  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 5701)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbrc.2016.09.005"; journalName: "Biochem Biophys Res 
            Commun"; date: "2016-09-30- 30"; volume: "478"; issue: "4"; pages: 
            "1694-9"
COMMENT     Sequence Label: pEGFP-C1-Nup35-mut
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             114..908
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             114..908
                     /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(168..187)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     778..797
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1096..1115)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1140..1187
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1516..2104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2005..2024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2279..2582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2583..2786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2737..2757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2825..2834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2831..2833,2834..2836,2837..3547)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2831..2833,2834..2836,2837..3547)
                     /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(2876..2897)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3137..3156)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3484..3505
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4707..4828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4744..4763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4798..4817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4835..5290)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4972..4993)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5184..5203
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5317..5421
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5423..5701,1..79)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5423..5614
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5446..5466)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5631..5701,1..65)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
       61 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
      121 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
      181 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
      241 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
      301 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
      361 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
      421 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
      481 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
      541 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
      601 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
      661 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
      721 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
      781 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
      841 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
      901 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
      961 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     1021 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     1081 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     1141 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     1201 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     1261 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     1321 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     1381 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1621 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     2221 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2281 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2341 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2401 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2461 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2521 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2581 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2641 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2701 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2761 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     2821 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2881 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2941 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3001 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3061 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3121 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3181 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3241 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3301 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3361 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3421 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3481 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3541 gtacaagtcc ggactcagat ctcgagacgc agcctttgca gtggaacctc aggggcccgc
     3601 gttaggatct gaaccaatga tgctgggttc acccacatct ccaaagccag gagttaatgc
     3661 ccagttctta cctggatttt taatggggga tttgccagct ccggtgactc cacaacctcg
     3721 atcaattagt ggcccttcag taggagtaat ggaaatgaga tcacctttac ttgcaggtgg
     3781 gtcaccacca caaccagttg taccagctca taaagataaa agtggcgctc caccagttag
     3841 aagtatatat gatgacattt ctagcccagg acttggatca acacctttaa cttcaagaag
     3901 acagccaaac atttcagtaa tgcagagtcc tcttgttgga gttacatcta ctcctggaac
     3961 agggcaaagt atgtttagtc cagcaagtat cggtcagcca cgaaagacga cattatctcc
     4021 tgcccagttg gatccttttt atactcaagg agattctttg acttcagaag atcacctcga
     4081 tgactcttgg gtgactgtat ttgggtttcc tcaagcatct gcttcctaca tattactaca
     4141 atttgcacag tatgggaata tcttaaaaca tgtgatgtct aatacaggaa attggatgca
     4201 tattcgttat caatctaaac tgcaggctcg gaaagcctta agcaaagatg ggaggatttt
     4261 tggagaatcc atcatgattg gtgtaaaacc atgtattgac aaaagtgtta tggaaagcag
     4321 tgacagatgt gctttatcat ctccatcttt agcctttaca ccaccaatca aaactctagg
     4381 tacaccaaca caacctggaa gtactcctag gatttctacc atgagacctc ttgctacagc
     4441 atacaaagcc tctactagtg attatcaggt tatttctgac agacaaacgc caaaaaaaga
     4501 tgaaagtctt gtatccaaag caatggagta catgtttggc tggtaggaat tctgcagtcg
     4561 acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     4621 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4681 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4741 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4801 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     4861 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4921 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4981 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5041 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5101 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5161 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5221 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5281 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5341 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5401 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     5461 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5521 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5581 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5641 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5701 c
//
