LOCUS       sequence-321675-        6291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 5 
            amino acids) at amino acid position 494.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6291)
  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: Nup54_mEGFP494(-5)
FEATURES             Location/Qualifiers
     source          1..6291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    599..679
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2200..2221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2461..2480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2620..2871
                     /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     2808..2829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3087..3208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3124..3143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3178..3197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3215..3670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3352..3373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3564..3583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3697..3801
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3803..4160
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3803..3994
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3826..3846)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4011..4146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4195..4989
                     /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             4195..4989
                     /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(4249..4268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4859..4878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5177..5196)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5221..5268
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5597..6185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6086..6105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
      661 gggcccggga tccaccggtc ccgcgacatg gcattcaatt ttggggctcc ttcgggcact
      721 tcgggcacct ctacagccac cgcggcaccg gcgggcgggt ttggcggatt tggaacaaca
      781 actacaactg caggctctgc attcagcttt tctgccccaa caaacacagg cagtacaggc
      841 cttctcggcg gcactcagaa caaaggtttt ggctttggca ctggttttgg cacatcgacg
      901 ggtactggca ctggtttagg cactggcttg ggaaccggac ttggattcgg aggatttaac
      961 acccagcagc agcagcagca gcagcagact tctttaggcg gtctcttcag tcagcctgca
     1021 caggcccctg cgcagtccaa ccaactcatc aacactgcca gcgctctttc tgctccaacg
     1081 ctattggggg atgagagaga tgccatcttg gcaaagtgga accagttgca ggccttctgg
     1141 ggaacaggaa aaggatattt caataataac attcctccag tggagtttac acaggaaaac
     1201 cccttttgcc gatttaaggc tgtcggttac agttgtatgc ccaataacaa agatgaagat
     1261 ggactagtgg ttctaatttt caacaaaaaa gaaacagata ttagaagcca acaacagcag
     1321 ttggtagaat cattgcataa agttctggga ggaaaccaga ccctcactgt caatgtggag
     1381 ggcattaaga cactgcccga cgatcagaca gaagttgtca tttacattgt tgagcgctct
     1441 ccaaatggca cctcgagaag agttccagcg acaacgctgt atgcccactt tgagcaagcc
     1501 aacattaaaa cacagcttca gcagcttggc gtgacccttt ctatgaccag aacagagctg
     1561 tctcctgctc agatcaagca gctcctacag aaccctcctg ctggtgttga tcccattatt
     1621 tgggaacagg ccaaggtgga taacccggat tctgaaaaat taattcctgt tccaatggtg
     1681 ggtttcaaag aattgcttcg aagactgaag gttcaagacc agatgactaa gcagcaccag
     1741 accagattag atatcatatc tgaagatatt agtgaactgc aaaagaatca aactacaact
     1801 atggccaaaa tagcacaata caagaggaaa ctgatggatc tctcacacag aaccttacag
     1861 gtcctaatca agcaggaaat ccagaggaag agtggctatg ccatccaggc tgaggaagaa
     1921 cagttgcggg ttcagctaga tacgattcaa ggagagctga atgcacccac tcagttcaaa
     1981 ggtcgactga atgaactgat gtcccagatc aggatgcaga atcattttgg agctgtgaaa
     2041 tctgaagaaa agtattacat agatgcagac ctattacgag agatcaagca gcatttgaaa
     2101 cagcaacagg aaggccttag ccacctgatt agcatcataa aagacgacct ggaagatatc
     2161 aagctggtag aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2221 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2281 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2341 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2401 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2461 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2521 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2581 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2641 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2701 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2761 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2821 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggagcatgga
     2881 ctgaatgaga ccatccacag cagaggcggg gtctttagct gaccattgcg ctgtccagag
     2941 gttcataaag ctgtgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     3001 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3061 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3121 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3181 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3241 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3661 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3721 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3781 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3841 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3961 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4021 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4081 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4141 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4201 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4261 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4321 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4381 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4441 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4501 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4561 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4621 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4681 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4741 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4801 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4861 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4921 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4981 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5041 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5101 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5161 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5221 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5281 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5341 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5401 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc g
//
