LOCUS       sequence-321672-        6309 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 6 
            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 6309)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6309)
  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(flex1)
FEATURES             Location/Qualifiers
     source          1..6309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    683..763
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2302..2323)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2563..2582)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2722..2973
                     /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     2910..2931
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3189..3310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3226..3245)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3280..3299
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3317..3772)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3454..3475)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3666..3685
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3799..3903
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3905..4262
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3905..4096
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3928..3948)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4113..4248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4297..5091
                     /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             4297..5091
                     /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(4351..4370)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4961..4980
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5279..5298)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5323..5370
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5699..6287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6188..6207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
      721 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcccgcga catggcattc
      781 aattttgggg ctccttcggg cacttcgggc acctctacag ccaccgcggc accggcgggc
      841 gggtttggcg gatttggaac aacaactaca actgcaggct ctgcattcag cttttctgcc
      901 ccaacaaaca caggcagtac aggccttctc ggcggcactc agaacaaagg ttttggcttt
      961 ggcactggtt ttggcacatc gacgggtact ggcactggtt taggcactgg cttgggaacc
     1021 ggacttggat tcggaggatt taacacccag cagcagcagc agcagcagca gacttcttta
     1081 ggcggtctct tcagtcagcc tgcacaggcc cctgcgcagt ccaaccaact catcaacact
     1141 gccagcgctc tttctgctcc aacgctattg ggggatgaga gagatgccat cttggcaaag
     1201 tggaaccagt tgcaggcctt ctggggaaca ggaaaaggat atttcaataa taacattcct
     1261 ccagtggagt ttacacagga aaaccccttt tgccgattta aggctgtcgg ttacagttgt
     1321 atgcccaata acaaagatga agatggacta gtggttctaa ttttcaacaa aaaagaaaca
     1381 gatattagaa gccaacaaca gcagttggta gaatcattgc ataaagttct gggaggaaac
     1441 cagaccctca ctgtcaatgt ggagggcatt aagacactgc ccgacgatca gacagaagtt
     1501 gtcatttaca ttgttgagcg ctctccaaat ggcacctcga gaagagttcc agcgacaacg
     1561 ctgtatgccc actttgagca agccaacatt aaaacacagc ttcagcagct tggcgtgacc
     1621 ctttctatga ccagaacaga gctgtctcct gctcagatca agcagctcct acagaaccct
     1681 cctgctggtg ttgatcccat tatttgggaa caggccaagg tggataaccc ggattctgaa
     1741 aaattaattc ctgttccaat ggtgggtttc aaagaattgc ttcgaagact gaaggttcaa
     1801 gaccagatga ctaagcagca ccagaccaga ttagatatca tatctgaaga tattagtgaa
     1861 ctgcaaaaga atcaaactac aactatggcc aaaatagcac aatacaagag gaaactgatg
     1921 gatctctcac acagaacctt acaggtccta atcaagcagg aaatccagag gaagagtggc
     1981 tatgccatcc aggctgagga agaacagttg cgggttcagc tagatacgat tcaaggagag
     2041 ctgaatgcac ccactcagtt caaaggtcga ctgaatgaac tgatgtccca gatcaggatg
     2101 cagaatcatt ttggagctgt gaaatctgaa gaaaagtatt acatagatgc agacctatta
     2161 cgagagatca agcagcattt gaaacagcaa caggaaggcc ttagccacct gattagcatc
     2221 ataaaagacg acctggaaga tatcaagctg gtagggggtg gcggcagctc cgaggagctg
     2281 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2341 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2401 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2461 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2521 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2581 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2641 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2701 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2761 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2821 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     2881 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2941 gggatcactc tcggcatgga cgagctgtac aaggagcatg gactgaatga gaccatccac
     3001 agcagaggcg gggtctttag ctgaccattg cgctgtccag aggttcataa agctgtgtac
     3061 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3121 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3181 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3301 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3361 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3421 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3481 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3541 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3601 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3661 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3721 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3781 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3841 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3901 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4081 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4141 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4201 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4261 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4321 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4381 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4441 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4501 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4561 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4621 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4681 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4741 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4801 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4861 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4921 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4981 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5041 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5101 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5161 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5221 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5281 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5341 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5401 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5461 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5521 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5581 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5641 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5701 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5761 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5821 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5881 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5941 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6001 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6061 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6301 cggcctttt
//
