LOCUS       sequence-325399-        5973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 without the FG-Nup domain, with mEGFP 
            (missing the first 5 amino acids) at amino acid position 494 with a 
            rigid alpha helix of 7 amino acids at the carboxyl end of mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  AUTHORS   Pulupa J, Prior H, Johnson DS, Simon SM
  TITLE     Conformation of the nuclear pore in living cells is modulated by 
            transport state.
  JOURNAL   Elife. 2020 Dec 21;9. pii: 60654. doi: 10.7554/eLife.60654.
  PUBMED    33346731
REFERENCE   2  (bases 1 to 5973)
  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"; journalName: "Elife."; 
            date: "2020-12-21"; volume: "9. pii: 60654. doi"; pages: " 
            10.7554/eLife.60654"
COMMENT     Sequence Label: Nup54 no FG-mEGFP494(2)
FEATURES             Location/Qualifiers
     source          1..5973
                     /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(1874..1895)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2135..2154)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2294..2545
                     /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     2482..2503
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2761..2882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2798..2817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2852..2871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2889..3344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3026..3047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3238..3257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3371..3475
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3477..3834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3477..3668
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3500..3520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3685..3820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3869..4663
                     /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             3869..4663
                     /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(3923..3942)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4533..4552
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4851..4870)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4895..4942
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5271..5859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5760..5779
                     /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 tggcgcagtc caaccaactc atcaacactg ccagcgctct
      721 ttctgctcca acgctattgg gggatgagag agatgccatc ttggcaaagt ggaaccagtt
      781 gcaggccttc tggggaacag gaaaaggata tttcaataat aacattcctc cagtggagtt
      841 tacacaggaa aacccctttt gccgatttaa ggctgtcggt tacagttgta tgcccaataa
      901 caaagatgaa gatggactag tggttctaat tttcaacaaa aaagaaacag atattagaag
      961 ccaacaacag cagttggtag aatcattgca taaagttctg ggaggaaacc agaccctcac
     1021 tgtcaatgtg gagggcatta agacactgcc cgacgatcag acagaagttg tcatttacat
     1081 tgttgagcgc tctccaaatg gcacctcgag aagagttcca gcgacaacgc tgtatgccca
     1141 ctttgagcaa gccaacatta aaacacagct tcagcagctt ggcgtgaccc tttctatgac
     1201 cagaacagag ctgtctcctg ctcagatcaa gcagctccta cagaaccctc ctgctggtgt
     1261 tgatcccatt atttgggaac aggccaaggt ggataacccg gattctgaaa aattaattcc
     1321 tgttccaatg gtgggtttca aagaattgct tcgaagactg aaggttcaag accagatgac
     1381 taagcagcac cagaccagat tagatatcat atctgaagat attagtgaac tgcaaaagaa
     1441 tcaaactaca actatggcca aaatagcaca atacaagagg aaactgatgg atctctcaca
     1501 cagaacctta caggtcctaa tcaagcagga aatccagagg aagagtggct atgccatcca
     1561 ggctgaggaa gaacagttgc gggttcagct agatacgatt caaggagagc tgaatgcacc
     1621 cactcagttc aaaggtcgac tgaatgaact gatgtcccag atcaggatgc agaatcattt
     1681 tggagctgtg aaatctgaag aaaagtatta catagatgca gacctattac gagagatcaa
     1741 gcagcatttg aaacagcaac aggaaggcct tagccacctg attagcatca taaaagacga
     1801 cctggaagat atcaagctgg tagcagaagc tgctgcaaag gaggaggagc tgttcaccgg
     1861 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1921 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1981 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2041 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2101 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2161 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2221 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2281 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2341 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2401 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     2461 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2521 tctcggcatg gacgagctgt acaaggagca tggactgaat gagaccatcc acagcagagg
     2581 cggggtcttt agctgaccat tgcgctgtcc agaggttcat aaagctgtgt acaagtaaag
     2641 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2701 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2881 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2941 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3001 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3061 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3121 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3181 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3241 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3301 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3361 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3421 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3481 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3781 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3841 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3901 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3961 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4021 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4081 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4141 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4201 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4261 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4321 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4381 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4441 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4501 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4561 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4621 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4681 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4741 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4801 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4861 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4921 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4981 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5041 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5101 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5161 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5221 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5281 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5341 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5401 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5461 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5521 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5581 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5641 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5701 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5761 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5821 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5881 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5941 gattctgtgg ataaccgtat taccgccatg cat
//
