LOCUS       sequence-316724-        6504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup58 with a mEGFP (missing the first 6 
            amino acids) at amino acid position 412.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6504)
  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(-6)
FEATURES             Location/Qualifiers
     source          1..6504
                     /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(1943..1964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2204..2223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2363..2614
                     /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     2551..2572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3292..3413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3329..3348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3383..3402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3420..3875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3557..3578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3769..3788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3902..4006
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4008..4365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4008..4199
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4031..4051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4216..4351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4400..5194
                     /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             4400..5194
                     /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(4454..4473)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5064..5083
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5382..5401)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5426..5473
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5802..6390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6291..6310
                     /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 tggccacggg gttctccttt gggtccggga ccctcggctc
      721 caccactgtg gcccccggcg gcaccggcac aggcagcgga ttctccttcg gcgcgtccag
      781 cacaccttct gtggggctca acttcggaac tctcggaagc tctgcgacac cagcatctac
      841 atctacttct gccagtggtt ttggaacagg actctttgga tctaagcctg gcactggatt
      901 cactctagga ggaacaagtg caggaacaac agctactaca tctgcatcta caacaggctt
      961 tagtttagga ttcagtaaac ctgcagcatc tgccacacca tttgccctgc ccgtcacttc
     1021 taccacagct agtggtctta ctctctcgtc tgctctgaca tcagctccag cagcatccac
     1081 aggatttacg ttgaataact tgggagcaac accagccaca actacagcag catcaacagg
     1141 actttcctta gggggggcct tagctggttt gggaggttca cttttccaga gtggaaacac
     1201 agcaacatca ggtctcggac agaatgcttt aagcttgtct ctggggactg caacacctac
     1261 ttcagcagcc agcagtgaag gccttggtgg tatagatttt agtacctcat cagataaaaa
     1321 gagtgataaa acgggaacaa ggccagagga tagcaaagcc ctgaaagatg aaaaccttcc
     1381 tcctgtcatc tgtcaggatg ttgagaatct ccagaaattt gtcaaggaac aaaaacaagt
     1441 ccaagaagaa attagtcgaa tgtcttccaa agcaatgctt aaagtccagg aggacatcaa
     1501 ggccctgaag cagctcctgt cattggctgc tagtgggttg cagaggaaca cactcaatat
     1561 tgacaaactg aagttagaaa ctgctcagga gttaaagaat gcggaaatag ctttaagaac
     1621 ccagaaaaca ccacctggac tccaacatga aaatacagct cctgctgact acttcagagt
     1681 cttggttcag caatttgagg tacagcttca gcagtacagg cagcagattg aagaactaga
     1741 aaaccatctt gccactcaag caaacaattc acatataact ccgcaagact tatcaatggc
     1801 tatgcagaaa atttatcaaa catttgtagc tttagctgca caacttcagt ccattcatga
     1861 aaatgtcaag gtgcttaaag aacagtatct cagctacaga aaaatgtttt tgggtgagct
     1921 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1981 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2041 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2101 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2161 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2221 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2281 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2341 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2401 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2461 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     2521 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2581 cgggatcact ctcggcatgg acgagctgta caaggatgtg tttgaagcca ggagaacaga
     2641 agccaagaag tggcagaatg cacccagagt tactactgga cccacgcctt tcagcaccat
     2701 gccaaacgca gcagccgttg ccatggctgc aacacttaca cagcagcaac agcctgctac
     2761 agggccacag ccgtctctgg gagttagttt tggaacgcca ttcggctcag gtattggcac
     2821 tggcttgcaa tcaagtggct taggttcttc aaaccttgga ggatttggaa ctagctctgg
     2881 tttcggatgc ggtaccacag gggcctccac atttggattt ggaacaacag ataaaccctc
     2941 aggaagtctt agtgcaggct ttggcagctc aagtacgtct gggtttaact tcagcaatcc
     3001 tggcatcacg gcatcagctg gtttgacatt tggggtgtcc aatcctgcgt ctgcggggtt
     3061 tggaacagga ggacaactcc ttcagttgaa gagacctccc gctggaaaca aaagaggaaa
     3121 gagatgacca ttgcgctgtc cagaggttca taaagctgtg tacaagtaaa gcggccgcga
     3181 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3241 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3301 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3361 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3421 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3481 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3541 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3601 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3661 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3721 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3781 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3841 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3901 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3961 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4021 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4201 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4261 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4321 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4381 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4441 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4501 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4561 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4621 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4681 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4741 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4801 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4861 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4921 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4981 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5041 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5101 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5161 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5221 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5281 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5341 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5401 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5461 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5521 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5581 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5641 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5941 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6481 gataaccgta ttaccgccat gcat
//
