LOCUS       sequence-321894-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup58 with a mEGFP (missing the first 7 
            amino acids) at amino acid position 412.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6501)
  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: Nup58_mEGFP(-7)
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    615..695
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2225..2244)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2384..2635
                     /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     2572..2593
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3313..3434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3350..3369)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3404..3423
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3441..3896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3578..3599)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3790..3809
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3923..4027
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4029..4386
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4029..4220
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4052..4072)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4237..4372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4421..5215
                     /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             4421..5215
                     /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(4475..4494)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5085..5104
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5403..5422)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5447..5494
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6312..6331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      601 gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct tcgaattctg
      661 cagtcgacgg taccgcgggc cctggatcca ccggtcccgc gacatggcca cggggttctc
      721 ctttgggtcc gggaccctcg gctccaccac tgtggccccc ggcggcaccg gcacaggcag
      781 cggattctcc ttcggcgcgt ccagcacacc ttctgtgggg ctcaacttcg gaactctcgg
      841 aagctctgcg acaccagcat ctacatctac ttctgccagt ggttttggaa caggactctt
      901 tggatctaag cctggcactg gattcactct aggaggaaca agtgcaggaa caacagctac
      961 tacatctgca tctacaacag gctttagttt aggattcagt aaacctgcag catctgccac
     1021 accatttgcc ctgcccgtca cttctaccac agctagtggt cttactctct cgtctgctct
     1081 gacatcagct ccagcagcat ccacaggatt tacgttgaat aacttgggag caacaccagc
     1141 cacaactaca gcagcatcaa caggactttc cttagggggg gccttagctg gtttgggagg
     1201 ttcacttttc cagagtggaa acacagcaac atcaggtctc ggacagaatg ctttaagctt
     1261 gtctctgggg actgcaacac ctacttcagc agccagcagt gaaggccttg gtggtataga
     1321 ttttagtacc tcatcagata aaaagagtga taaaacggga acaaggccag aggatagcaa
     1381 agccctgaaa gatgaaaacc ttcctcctgt catctgtcag gatgttgaga atctccagaa
     1441 atttgtcaag gaacaaaaac aagtccaaga agaaattagt cgaatgtctt ccaaagcaat
     1501 gcttaaagtc caggaggaca tcaaggccct gaagcagctc ctgtcattgg ctgctagtgg
     1561 gttgcagagg aacacactca atattgacaa actgaagtta gaaactgctc aggagttaaa
     1621 gaatgcggaa atagctttaa gaacccagaa aacaccacct ggactccaac atgaaaatac
     1681 agctcctgct gactacttca gagtcttggt tcagcaattt gaggtacagc ttcagcagta
     1741 caggcagcag attgaagaac tagaaaacca tcttgccact caagcaaaca attcacatat
     1801 aactccgcaa gacttatcaa tggctatgca gaaaatttat caaacatttg tagctttagc
     1861 tgcacaactt cagtccattc atgaaaatgt caaggtgctt aaagaacagt atctcagcta
     1921 cagaaaaatg tttgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1981 tgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2041 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2101 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2161 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2221 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2281 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2341 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2401 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2461 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2521 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2581 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggatgt
     2641 gtttgaagcc aggagaacag aagccaagaa gtggcagaat gcacccagag ttactactgg
     2701 acccacgcct ttcagcacca tgccaaacgc agcagccgtt gccatggctg caacacttac
     2761 acagcagcaa cagcctgcta cagggccaca gccgtctctg ggagttagtt ttggaacgcc
     2821 attcggctca ggtattggca ctggcttgca atcaagtggc ttaggttctt caaaccttgg
     2881 aggatttgga actagctctg gtttcggatg cggtaccaca ggggcctcca catttggatt
     2941 tggaacaaca gataaaccct caggaagtct tagtgcaggc tttggcagct caagtacgtc
     3001 tgggtttaac ttcagcaatc ctggcatcac ggcatcagct ggtttgacat ttggggtgtc
     3061 caatcctgcg tctgcggggt ttggaacagg aggacaactc cttcagttga agagacctcc
     3121 cgctggaaac aaaagaggaa agagatgacc attgcgctgt ccagaggttc ataaagctgt
     3181 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     3241 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3301 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3361 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3421 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3481 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3541 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3601 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3661 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3721 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3781 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3841 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3901 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3961 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4021 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4201 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4261 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4321 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4381 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4441 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4501 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4561 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4621 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4681 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4741 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4801 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4861 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4921 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4981 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5041 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5101 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5161 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5221 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5281 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5341 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     5401 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     5461 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5521 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5581 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5641 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5941 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6481 gcgttatccc ctgattctgt g
//
