LOCUS       sequence-321833-        6246 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 6246)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6246)
  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..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    648..728
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2294..2315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2555..2574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2714..2965
                     /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     2902..2923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3091..3212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3128..3147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3182..3201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3219..3674)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3356..3377)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3568..3587
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3701..3805
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3807..4164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3807..3998
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3830..3850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4015..4150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4199..4993
                     /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             4199..4993
                     /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(4253..4272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4863..4882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5181..5200)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5225..5272
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
      661 gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
      721 ccaccggtcc cgcgacatgg cattcaattt tggggctcct tcgggcactt cgggcacctc
      781 tacagccacc gcggcaccgg cgggcgggtt tggcggattt ggaacaacaa ctacaactgc
      841 aggctctgca ttcagctttt ctgccccaac aaacacaggc agtacaggcc ttctcggcgg
      901 cactcagaac aaaggttttg gctttggcac tggttttggc acatcgacgg gtactggcac
      961 tggtttaggc actggcttgg gaaccggact tggattcgga ggatttaaca cccagcagca
     1021 gcagcagcag cagcagactt ctttaggcgg tctcttcagt cagcctgcac aggcccctgc
     1081 gcagtccaac caactcatca acactgccag cgctctttct gctccaacgc tattggggga
     1141 tgagagagat gccatcttgg caaagtggaa ccagttgcag gccttctggg gaacaggaaa
     1201 aggatatttc aataataaca ttcctccagt ggagtttaca caggaaaacc ccttttgccg
     1261 atttaaggct gtcggttaca gttgtatgcc caataacaaa gatgaagatg gactagtggt
     1321 tctaattttc aacaaaaaag aaacagatat tagaagccaa caacagcagt tggtagaatc
     1381 attgcataaa gttctgggag gaaaccagac cctcactgtc aatgtggagg gcattaagac
     1441 actgcccgac gatcagacag aagttgtcat ttacattgtt gagcgctctc caaatggcac
     1501 ctcgagaaga gttccagcga caacgctgta tgcccacttt gagcaagcca acattaaaac
     1561 acagcttcag cagcttggcg tgaccctttc tatgaccaga acagagctgt ctcctgctca
     1621 gatcaagcag ctcctacaga accctcctgc tggtgttgat cccattattt gggaacaggc
     1681 caaggtggat aacccggatt ctgaaaaatt aattcctgtt ccaatggtgg gtttcaaaga
     1741 attgcttcga agactgaagg ttcaagacca gatgactaag cagcaccaga ccagattaga
     1801 tatcatatct gaagatatta gtgaactgca aaagaatcaa actacaacta tggccaaaat
     1861 agcacaatac aagaggaaac tgatggatct ctcacacaga accttacagg tcctaatcaa
     1921 gcaggaaatc cagaggaaga gtggctatgc catccaggct gaggaagaac agttgcgggt
     1981 tcagctagat acgattcaag gagagctgaa tgcacccact cagttcaaag gtcgactgaa
     2041 tgaactgatg tcccagatca ggatgcagaa tcattttgga gctgtgaaat ctgaagaaaa
     2101 gtattacata gatgcagacc tattacgaga gatcaagcag catttgaaac agcaacagga
     2161 aggccttagc cacctgatta gcatcataaa agacgacctg gaagatatca agctggtaga
     2221 gcatggactg aatgagacca tccacagcag aggcggggtc tttagcgagc tgttcaccgg
     2281 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2341 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2401 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2461 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2521 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2581 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2641 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2701 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2761 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2821 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     2881 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2941 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     3001 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3061 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3121 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3181 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     3241 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3301 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3361 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3421 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3481 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3541 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3601 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3661 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3781 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4141 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4201 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4261 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4321 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4381 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4441 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4501 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4561 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4621 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4681 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4741 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4801 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4861 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4921 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4981 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5041 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5101 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5161 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5221 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5281 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5341 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5401 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcac
//
