LOCUS       sequence-321674-        6294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 4 
            amino acids) at amino acid position 494.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6294)
  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(-4)
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    607..687
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2211..2232)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2472..2491)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2631..2882
                     /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     2819..2840
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3098..3219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3135..3154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3189..3208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3226..3681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3363..3384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3575..3594
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3708..3812
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3814..4171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3814..4005
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3837..3857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4022..4157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4206..5000
                     /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             4206..5000
                     /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(4260..4279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4870..4889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5188..5207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5232..5279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5608..6196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6097..6116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac
      661 ggtaccgcgg gcccgggatc caccggtccc gcgacatggc attcaatttt ggggctcctt
      721 cgggcacttc gggcacctct acagccaccg cggcaccggc gggcgggttt ggcggatttg
      781 gaacaacaac tacaactgca ggctctgcat tcagcttttc tgccccaaca aacacaggca
      841 gtacaggcct tctcggcggc actcagaaca aaggttttgg ctttggcact ggttttggca
      901 catcgacggg tactggcact ggtttaggca ctggcttggg aaccggactt ggattcggag
      961 gatttaacac ccagcagcag cagcagcagc agcagacttc tttaggcggt ctcttcagtc
     1021 agcctgcaca ggcccctgcg cagtccaacc aactcatcaa cactgccagc gctctttctg
     1081 ctccaacgct attgggggat gagagagatg ccatcttggc aaagtggaac cagttgcagg
     1141 ccttctgggg aacaggaaaa ggatatttca ataataacat tcctccagtg gagtttacac
     1201 aggaaaaccc cttttgccga tttaaggctg tcggttacag ttgtatgccc aataacaaag
     1261 atgaagatgg actagtggtt ctaattttca acaaaaaaga aacagatatt agaagccaac
     1321 aacagcagtt ggtagaatca ttgcataaag ttctgggagg aaaccagacc ctcactgtca
     1381 atgtggaggg cattaagaca ctgcccgacg atcagacaga agttgtcatt tacattgttg
     1441 agcgctctcc aaatggcacc tcgagaagag ttccagcgac aacgctgtat gcccactttg
     1501 agcaagccaa cattaaaaca cagcttcagc agcttggcgt gaccctttct atgaccagaa
     1561 cagagctgtc tcctgctcag atcaagcagc tcctacagaa ccctcctgct ggtgttgatc
     1621 ccattatttg ggaacaggcc aaggtggata acccggattc tgaaaaatta attcctgttc
     1681 caatggtggg tttcaaagaa ttgcttcgaa gactgaaggt tcaagaccag atgactaagc
     1741 agcaccagac cagattagat atcatatctg aagatattag tgaactgcaa aagaatcaaa
     1801 ctacaactat ggccaaaata gcacaataca agaggaaact gatggatctc tcacacagaa
     1861 ccttacaggt cctaatcaag caggaaatcc agaggaagag tggctatgcc atccaggctg
     1921 aggaagaaca gttgcgggtt cagctagata cgattcaagg agagctgaat gcacccactc
     1981 agttcaaagg tcgactgaat gaactgatgt cccagatcag gatgcagaat cattttggag
     2041 ctgtgaaatc tgaagaaaag tattacatag atgcagacct attacgagag atcaagcagc
     2101 atttgaaaca gcaacaggaa ggccttagcc acctgattag catcataaaa gacgacctgg
     2161 aagatatcaa gctggtaggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2221 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2281 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2341 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2401 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2461 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2521 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2581 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2641 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2701 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2761 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     2821 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2881 aggagcatgg actgaatgag accatccaca gcagaggcgg ggtctttagc tgaccattgc
     2941 gctgtccaga ggttcataaa gctgtgtaca agtaaagcgg ccgcgactct agatcataat
     3001 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3061 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3121 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3181 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3241 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3301 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3361 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3421 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3481 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3541 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3601 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3661 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3781 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3841 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3901 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3961 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4021 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4081 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4141 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4201 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4261 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4321 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4381 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4441 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4501 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4561 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4621 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4681 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4741 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4801 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4861 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4921 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4981 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5041 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5101 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5161 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5221 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5281 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5341 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5401 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5461 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5521 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accg
//
