LOCUS       sequence-316737-        6294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 4 
            amino acids) at the caroxyl-terminus with no linker and a total net 
            fusion of (-4) amino acids.
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_mEGFP510(-4)
FEATURES             Location/Qualifiers
     source          1..6294
                     /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"
     CDS             join(2201..2203,2204..2917)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VFSGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2243..2264)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2504..2523)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2851..2872
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3082..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3119..3138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3173..3192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3210..3665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3347..3368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3559..3578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3692..3796
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3798..4155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3798..3989
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3821..3841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4006..4141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4190..4984
                     /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             4190..4984
                     /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(4244..4263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4854..4873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5172..5191)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5216..5263
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5592..6180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6081..6100
                     /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 tggcattcaa ttttggggct ccttcgggca cttcgggcac
      721 ctctacagcc accgcggcac cggcgggcgg gtttggcgga tttggaacaa caactacaac
      781 tgcaggctct gcattcagct tttctgcccc aacaaacaca ggcagtacag gccttctcgg
      841 cggcactcag aacaaaggtt ttggctttgg cactggtttt ggcacatcga cgggtactgg
      901 cactggttta ggcactggct tgggaaccgg acttggattc ggaggattta acacccagca
      961 gcagcagcag cagcagcaga cttctttagg cggtctcttc agtcagcctg cacaggcccc
     1021 tgcgcagtcc aaccaactca tcaacactgc cagcgctctt tctgctccaa cgctattggg
     1081 ggatgagaga gatgccatct tggcaaagtg gaaccagttg caggccttct ggggaacagg
     1141 aaaaggatat ttcaataata acattcctcc agtggagttt acacaggaaa accccttttg
     1201 ccgatttaag gctgtcggtt acagttgtat gcccaataac aaagatgaag atggactagt
     1261 ggttctaatt ttcaacaaaa aagaaacaga tattagaagc caacaacagc agttggtaga
     1321 atcattgcat aaagttctgg gaggaaacca gaccctcact gtcaatgtgg agggcattaa
     1381 gacactgccc gacgatcaga cagaagttgt catttacatt gttgagcgct ctccaaatgg
     1441 cacctcgaga agagttccag cgacaacgct gtatgcccac tttgagcaag ccaacattaa
     1501 aacacagctt cagcagcttg gcgtgaccct ttctatgacc agaacagagc tgtctcctgc
     1561 tcagatcaag cagctcctac agaaccctcc tgctggtgtt gatcccatta tttgggaaca
     1621 ggccaaggtg gataacccgg attctgaaaa attaattcct gttccaatgg tgggtttcaa
     1681 agaattgctt cgaagactga aggttcaaga ccagatgact aagcagcacc agaccagatt
     1741 agatatcata tctgaagata ttagtgaact gcaaaagaat caaactacaa ctatggccaa
     1801 aatagcacaa tacaagagga aactgatgga tctctcacac agaaccttac aggtcctaat
     1861 caagcaggaa atccagagga agagtggcta tgccatccag gctgaggaag aacagttgcg
     1921 ggttcagcta gatacgattc aaggagagct gaatgcaccc actcagttca aaggtcgact
     1981 gaatgaactg atgtcccaga tcaggatgca gaatcatttt ggagctgtga aatctgaaga
     2041 aaagtattac atagatgcag acctattacg agagatcaag cagcatttga aacagcaaca
     2101 ggaaggcctt agccacctga ttagcatcat aaaagacgac ctggaagata tcaagctggt
     2161 agagcatgga ctgaatgaga ccatccacag cagaggcggg gtctttagcg gcgaggagct
     2221 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2281 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2341 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2401 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2461 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2521 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2581 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2641 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2701 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2761 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     2821 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2881 cgggatcact ctcggcatgg acgagctgta caagtgacca ttgcgctgtc cagaggttca
     2941 taaagctgtg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     3001 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3061 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3121 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3181 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     3241 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3301 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3361 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3421 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3481 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3541 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3601 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3661 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3721 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3781 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3841 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3901 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3961 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4021 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4081 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4141 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4201 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4261 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4321 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4381 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4441 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4501 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4561 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4621 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4681 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4741 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4801 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4861 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4921 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4981 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5041 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5101 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5161 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5221 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5281 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5341 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5401 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5461 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5521 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5581 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5641 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5701 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5761 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5821 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5881 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5941 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6001 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6061 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6121 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6181 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6241 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
