LOCUS       sequence-321665-        6312 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 with a rigid alpha helix of 
            7 amino acids at the carboxyl end of the mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6312)
  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(2)
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    579..659
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2201..2222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2462..2481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2621..2872
                     /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     2809..2830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3088..3209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3125..3144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3179..3198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3216..3671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3353..3374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3565..3584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3698..3802
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3804..4161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3804..3995
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3827..3847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4012..4147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4196..4990
                     /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             4196..4990
                     /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(4250..4269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4860..4879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5178..5197)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5222..5269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5598..6186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6087..6106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
      601 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc
      661 ccgcgacatg gcattcaatt ttggggctcc ttcgggcact tcgggcacct ctacagccac
      721 cgcggcaccg gcgggcgggt ttggcggatt tggaacaaca actacaactg caggctctgc
      781 attcagcttt tctgccccaa caaacacagg cagtacaggc cttctcggcg gcactcagaa
      841 caaaggtttt ggctttggca ctggttttgg cacatcgacg ggtactggca ctggtttagg
      901 cactggcttg ggaaccggac ttggattcgg aggatttaac acccagcagc agcagcagca
      961 gcagcagact tctttaggcg gtctcttcag tcagcctgca caggcccctg cgcagtccaa
     1021 ccaactcatc aacactgcca gcgctctttc tgctccaacg ctattggggg atgagagaga
     1081 tgccatcttg gcaaagtgga accagttgca ggccttctgg ggaacaggaa aaggatattt
     1141 caataataac attcctccag tggagtttac acaggaaaac cccttttgcc gatttaaggc
     1201 tgtcggttac agttgtatgc ccaataacaa agatgaagat ggactagtgg ttctaatttt
     1261 caacaaaaaa gaaacagata ttagaagcca acaacagcag ttggtagaat cattgcataa
     1321 agttctggga ggaaaccaga ccctcactgt caatgtggag ggcattaaga cactgcccga
     1381 cgatcagaca gaagttgtca tttacattgt tgagcgctct ccaaatggca cctcgagaag
     1441 agttccagcg acaacgctgt atgcccactt tgagcaagcc aacattaaaa cacagcttca
     1501 gcagcttggc gtgacccttt ctatgaccag aacagagctg tctcctgctc agatcaagca
     1561 gctcctacag aaccctcctg ctggtgttga tcccattatt tgggaacagg ccaaggtgga
     1621 taacccggat tctgaaaaat taattcctgt tccaatggtg ggtttcaaag aattgcttcg
     1681 aagactgaag gttcaagacc agatgactaa gcagcaccag accagattag atatcatatc
     1741 tgaagatatt agtgaactgc aaaagaatca aactacaact atggccaaaa tagcacaata
     1801 caagaggaaa ctgatggatc tctcacacag aaccttacag gtcctaatca agcaggaaat
     1861 ccagaggaag agtggctatg ccatccaggc tgaggaagaa cagttgcggg ttcagctaga
     1921 tacgattcaa ggagagctga atgcacccac tcagttcaaa ggtcgactga atgaactgat
     1981 gtcccagatc aggatgcaga atcattttgg agctgtgaaa tctgaagaaa agtattacat
     2041 agatgcagac ctattacgag agatcaagca gcatttgaaa cagcaacagg aaggccttag
     2101 ccacctgatt agcatcataa aagacgacct ggaagatatc aagctggtag cagaagctgc
     2161 tgcaaaggag gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2221 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2281 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2341 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2401 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2461 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2521 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2581 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2641 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2701 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2761 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2821 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca aggagcatgg
     2881 actgaatgag accatccaca gcagaggcgg ggtctttagc tgaccattgc gctgtccaga
     2941 ggttcataaa gctgtgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     3001 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3061 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3121 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3181 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3241 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3301 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3361 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3421 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3481 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3541 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3601 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3661 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3721 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3781 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3841 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3901 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4021 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4081 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4141 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4201 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4261 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4321 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4381 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4441 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4501 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4561 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4621 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4681 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4741 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4801 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4861 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4921 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4981 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5041 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5101 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5161 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5221 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5281 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5341 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5401 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5461 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5521 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     6301 tagttattaa ta
//
