LOCUS       sequence-316726-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup58 with a mEGFP (missing the first 8 
            amino acids) at amino acid position 412.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6498)
  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"
COMMENT     Sequence Label: Nup58_mEGFP(-8)
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    591..671
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1937..1958)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2198..2217)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2357..2608
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     2545..2566
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3286..3407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3323..3342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3377..3396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3414..3869)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3551..3572)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3763..3782
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3896..4000
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4002..4359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4002..4193
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4025..4045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4210..4345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4394..5188
                     /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             4394..5188
                     /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(4448..4467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5058..5077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5376..5395)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5420..5467
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5796..6384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6285..6304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcccgcgaca tggccacggg gttctccttt gggtccggga ccctcggctc
      721 caccactgtg gcccccggcg gcaccggcac aggcagcgga ttctccttcg gcgcgtccag
      781 cacaccttct gtggggctca acttcggaac tctcggaagc tctgcgacac cagcatctac
      841 atctacttct gccagtggtt ttggaacagg actctttgga tctaagcctg gcactggatt
      901 cactctagga ggaacaagtg caggaacaac agctactaca tctgcatcta caacaggctt
      961 tagtttagga ttcagtaaac ctgcagcatc tgccacacca tttgccctgc ccgtcacttc
     1021 taccacagct agtggtctta ctctctcgtc tgctctgaca tcagctccag cagcatccac
     1081 aggatttacg ttgaataact tgggagcaac accagccaca actacagcag catcaacagg
     1141 actttcctta gggggggcct tagctggttt gggaggttca cttttccaga gtggaaacac
     1201 agcaacatca ggtctcggac agaatgcttt aagcttgtct ctggggactg caacacctac
     1261 ttcagcagcc agcagtgaag gccttggtgg tatagatttt agtacctcat cagataaaaa
     1321 gagtgataaa acgggaacaa ggccagagga tagcaaagcc ctgaaagatg aaaaccttcc
     1381 tcctgtcatc tgtcaggatg ttgagaatct ccagaaattt gtcaaggaac aaaaacaagt
     1441 ccaagaagaa attagtcgaa tgtcttccaa agcaatgctt aaagtccagg aggacatcaa
     1501 ggccctgaag cagctcctgt cattggctgc tagtgggttg cagaggaaca cactcaatat
     1561 tgacaaactg aagttagaaa ctgctcagga gttaaagaat gcggaaatag ctttaagaac
     1621 ccagaaaaca ccacctggac tccaacatga aaatacagct cctgctgact acttcagagt
     1681 cttggttcag caatttgagg tacagcttca gcagtacagg cagcagattg aagaactaga
     1741 aaaccatctt gccactcaag caaacaattc acatataact ccgcaagact tatcaatggc
     1801 tatgcagaaa atttatcaaa catttgtagc tttagctgca caacttcagt ccattcatga
     1861 aaatgtcaag gtgcttaaag aacagtatct cagctacaga aaaatggagg agctgttcac
     1921 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1981 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2041 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2101 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2161 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2221 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2281 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2341 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2401 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2461 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     2521 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2581 cactctcggc atggacgagc tgtacaagga tgtgtttgaa gccaggagaa cagaagccaa
     2641 gaagtggcag aatgcaccca gagttactac tggacccacg cctttcagca ccatgccaaa
     2701 cgcagcagcc gttgccatgg ctgcaacact tacacagcag caacagcctg ctacagggcc
     2761 acagccgtct ctgggagtta gttttggaac gccattcggc tcaggtattg gcactggctt
     2821 gcaatcaagt ggcttaggtt cttcaaacct tggaggattt ggaactagct ctggtttcgg
     2881 atgcggtacc acaggggcct ccacatttgg atttggaaca acagataaac cctcaggaag
     2941 tcttagtgca ggctttggca gctcaagtac gtctgggttt aacttcagca atcctggcat
     3001 cacggcatca gctggtttga catttggggt gtccaatcct gcgtctgcgg ggtttggaac
     3061 aggaggacaa ctccttcagt tgaagagacc tcccgctgga aacaaaagag gaaagagatg
     3121 accattgcgc tgtccagagg ttcataaagc tgtgtacaag taaagcggcc gcgactctag
     3181 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3241 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3301 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3361 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3421 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3481 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3541 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3601 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3661 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3721 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3781 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3841 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3901 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3961 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4021 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4381 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4441 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4501 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4561 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4621 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4681 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4741 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4801 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4861 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4921 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4981 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5041 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5101 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5161 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5221 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5281 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5341 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5401 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5461 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5521 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5581 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5641 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5701 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5941 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6481 cgtattaccg ccatgcat
//
