LOCUS       sequence-316738-        6291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 5 
            amino acids) at the caroxyl-terminus with no linker and a total net 
            fusion of (-5) amino acids.
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_mEGFP510(-5)
FEATURES             Location/Qualifiers
     source          1..6291
                     /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(2240..2261)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2501..2520)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2660..2911
                     /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     2848..2869
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3079..3200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3116..3135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3170..3189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3207..3662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3344..3365)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3556..3575
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3689..3793
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3795..4152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3795..3986
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3818..3838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4003..4138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4187..4981
                     /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             4187..4981
                     /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(4241..4260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4851..4870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5169..5188)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5213..5260
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5589..6177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6078..6097
                     /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 aggagctgtt
     2221 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2281 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2341 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2401 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2461 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2521 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2581 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2641 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2701 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2761 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     2821 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2881 gatcactctc ggcatggacg agctgtacaa gtgaccattg cgctgtccag aggttcataa
     2941 agctgtgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3001 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3061 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3181 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3241 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3301 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3361 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3421 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3481 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3541 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3601 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3661 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3721 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3781 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3961 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4021 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4081 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4141 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4201 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4261 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4321 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4381 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4441 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4501 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4561 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4621 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4681 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4741 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4801 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4861 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4921 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4981 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5041 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5101 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5161 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5221 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5281 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5341 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5401 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5701 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
