LOCUS       sequence-316739-        6288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 6 
            amino acids) at the caroxyl-terminus with no linker and a total net 
            fusion of (-6) amino acids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6288)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6288)
  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(-6)
FEATURES             Location/Qualifiers
     source          1..6288
                     /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(2237..2258)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2498..2517)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2657..2908
                     /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     2845..2866
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3076..3197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3113..3132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3167..3186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3204..3659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3341..3362)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3553..3572
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3686..3790
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3792..4149
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3792..3983
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3815..3835)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4000..4135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4184..4978
                     /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             4184..4978
                     /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(4238..4257)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4848..4867
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5166..5185)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5210..5257
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5586..6174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6075..6094
                     /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 agctgttcac
     2221 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2281 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2341 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2401 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2461 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2521 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2581 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2641 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2701 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2761 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     2821 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2881 cactctcggc atggacgagc tgtacaagtg accattgcgc tgtccagagg ttcataaagc
     2941 tgtgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     3001 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3061 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3181 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3361 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3421 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3481 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3541 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3601 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3661 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3721 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3781 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3961 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4021 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4081 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4141 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4201 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4261 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4321 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4381 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4441 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4501 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4561 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4621 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4681 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4741 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4801 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4861 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4921 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4981 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5041 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5101 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5161 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5221 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5281 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5341 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5401 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5461 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5521 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5581 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5641 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5701 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5761 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5821 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5881 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5941 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6001 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6061 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6121 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6181 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6241 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
