LOCUS       sequence-321671-        6306 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 flexible linker of 5 
            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 6306)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6306)
  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(flex0)
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        31..334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        335..538
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     489..509
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    561..641
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2177..2198)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2438..2457)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2597..2848
                     /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     2785..2806
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3064..3185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3155..3174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3192..3647)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3329..3350)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3541..3560
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3780..4137
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3780..3971
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3803..3823)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3988..4123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4172..4966
                     /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             4172..4966
                     /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(4226..4245)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4836..4855
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5154..5173)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5198..5245
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5574..6162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6063..6082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
       61 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      121 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      181 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      241 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      301 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      361 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      421 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      481 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      541 tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttcga
      601 attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcccgcgaca tggcattcaa
      661 ttttggggct ccttcgggca cttcgggcac ctctacagcc accgcggcac cggcgggcgg
      721 gtttggcgga tttggaacaa caactacaac tgcaggctct gcattcagct tttctgcccc
      781 aacaaacaca ggcagtacag gccttctcgg cggcactcag aacaaaggtt ttggctttgg
      841 cactggtttt ggcacatcga cgggtactgg cactggttta ggcactggct tgggaaccgg
      901 acttggattc ggaggattta acacccagca gcagcagcag cagcagcaga cttctttagg
      961 cggtctcttc agtcagcctg cacaggcccc tgcgcagtcc aaccaactca tcaacactgc
     1021 cagcgctctt tctgctccaa cgctattggg ggatgagaga gatgccatct tggcaaagtg
     1081 gaaccagttg caggccttct ggggaacagg aaaaggatat ttcaataata acattcctcc
     1141 agtggagttt acacaggaaa accccttttg ccgatttaag gctgtcggtt acagttgtat
     1201 gcccaataac aaagatgaag atggactagt ggttctaatt ttcaacaaaa aagaaacaga
     1261 tattagaagc caacaacagc agttggtaga atcattgcat aaagttctgg gaggaaacca
     1321 gaccctcact gtcaatgtgg agggcattaa gacactgccc gacgatcaga cagaagttgt
     1381 catttacatt gttgagcgct ctccaaatgg cacctcgaga agagttccag cgacaacgct
     1441 gtatgcccac tttgagcaag ccaacattaa aacacagctt cagcagcttg gcgtgaccct
     1501 ttctatgacc agaacagagc tgtctcctgc tcagatcaag cagctcctac agaaccctcc
     1561 tgctggtgtt gatcccatta tttgggaaca ggccaaggtg gataacccgg attctgaaaa
     1621 attaattcct gttccaatgg tgggtttcaa agaattgctt cgaagactga aggttcaaga
     1681 ccagatgact aagcagcacc agaccagatt agatatcata tctgaagata ttagtgaact
     1741 gcaaaagaat caaactacaa ctatggccaa aatagcacaa tacaagagga aactgatgga
     1801 tctctcacac agaaccttac aggtcctaat caagcaggaa atccagagga agagtggcta
     1861 tgccatccag gctgaggaag aacagttgcg ggttcagcta gatacgattc aaggagagct
     1921 gaatgcaccc actcagttca aaggtcgact gaatgaactg atgtcccaga tcaggatgca
     1981 gaatcatttt ggagctgtga aatctgaaga aaagtattac atagatgcag acctattacg
     2041 agagatcaag cagcatttga aacagcaaca ggaaggcctt agccacctga ttagcatcat
     2101 aaaagacgac ctggaagata tcaagctggt agggggtggc ggcagcgagg agctgttcac
     2161 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2221 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2281 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2341 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2401 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2461 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2521 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2581 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2641 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2701 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     2761 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2821 cactctcggc atggacgagc tgtacaagga gcatggactg aatgagacca tccacagcag
     2881 aggcggggtc tttagctgac cattgcgctg tccagaggtt cataaagctg tgtacaagta
     2941 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     3001 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3061 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3121 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3181 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3241 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3301 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3361 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3421 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3481 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3541 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3601 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3781 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3841 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3961 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4021 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4081 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4141 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4201 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4261 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4321 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4381 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4441 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4501 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4561 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4621 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4681 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4741 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4801 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4861 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4921 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4981 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5041 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5101 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5161 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5221 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5281 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5341 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5401 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6241 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     6301 ggggtc
//
