LOCUS       sequence-321891-        6477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup58 with a mEGFP (missing the first 8 
            amino acids) at amino acid position 412.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6477)
  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(-8)
FEATURES             Location/Qualifiers
     source          1..6477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        29..332
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        333..536
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     487..507
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    559..639
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2166..2185)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2325..2576
                     /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     2513..2534
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3254..3375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3291..3310)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3345..3364
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3382..3837)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3519..3540)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3731..3750
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3864..3968
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3970..4161
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3993..4013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4158..4292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4341..5135
                     /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             4341..5135
                     /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(4395..4414)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5005..5024
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5323..5342)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5367..5414
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5743..6331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6232..6251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
       61 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      121 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      181 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      241 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      301 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      361 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      421 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      481 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      541 tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat
      601 tctgcagtcg acggtaccgc gggccctgga tccaccggtc ccgcgacatg gccacggggt
      661 tctcctttgg gtccgggacc ctcggctcca ccactgtggc ccccggcggc accggcacag
      721 gcagcggatt ctccttcggc gcgtccagca caccttctgt ggggctcaac ttcggaactc
      781 tcggaagctc tgcgacacca gcatctacat ctacttctgc cagtggtttt ggaacaggac
      841 tctttggatc taagcctggc actggattca ctctaggagg aacaagtgca ggaacaacag
      901 ctactacatc tgcatctaca acaggcttta gtttaggatt cagtaaacct gcagcatctg
      961 ccacaccatt tgccctgccc gtcacttcta ccacagctag tggtcttact ctctcgtctg
     1021 ctctgacatc agctccagca gcatccacag gatttacgtt gaataacttg ggagcaacac
     1081 cagccacaac tacagcagca tcaacaggac tttccttagg gggggcctta gctggtttgg
     1141 gaggttcact tttccagagt ggaaacacag caacatcagg tctcggacag aatgctttaa
     1201 gcttgtctct ggggactgca acacctactt cagcagccag cagtgaaggc cttggtggta
     1261 tagattttag tacctcatca gataaaaaga gtgataaaac gggaacaagg ccagaggata
     1321 gcaaagccct gaaagatgaa aaccttcctc ctgtcatctg tcaggatgtt gagaatctcc
     1381 agaaatttgt caaggaacaa aaacaagtcc aagaagaaat tagtcgaatg tcttccaaag
     1441 caatgcttaa agtccaggag gacatcaagg ccctgaagca gctcctgtca ttggctgcta
     1501 gtgggttgca gaggaacaca ctcaatattg acaaactgaa gttagaaact gctcaggagt
     1561 taaagaatgc ggaaatagct ttaagaaccc agaaaacacc acctggactc caacatgaaa
     1621 atacagctcc tgctgactac ttcagagtct tggttcagca atttgaggta cagcttcagc
     1681 agtacaggca gcagattgaa gaactagaaa accatcttgc cactcaagca aacaattcac
     1741 atataactcc gcaagactta tcaatggcta tgcagaaaat ttatcaaaca tttgtagctt
     1801 tagctgcaca acttcagtcc attcatgaaa atgtcaaggt gcttaaagaa cagtatctca
     1861 gctacagaaa aatggaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1921 gtgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1981 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2041 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2101 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2161 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2221 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2281 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2341 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2401 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2461 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2521 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggatg
     2581 tgtttgaagc caggagaaca gaagccaaga agtggcagaa tgcacccaga gttactactg
     2641 gacccacgcc tttcagcacc atgccaaacg cagcagccgt tgccatggct gcaacactta
     2701 cacagcagca acagcctgct acagggccac agccgtctct gggagttagt tttggaacgc
     2761 cattcggctc aggtattggc actggcttgc aatcaagtgg cttaggttct tcaaaccttg
     2821 gaggatttgg aactagctct ggtttcggat gcggtaccac aggggcctcc acatttggat
     2881 ttggaacaac agataaaccc tcaggaagtc ttagtgcagg ctttggcagc tcaagtacgt
     2941 ctgggtttaa cttcagcaat cctggcatca cggcatcagc tggtttgaca tttggggtgt
     3001 ccaatcctgc gtctgcgggg tttggaacag gaggacaact ccttcagttg aagagacctc
     3061 ccgctggaaa caaaagagga aagagatgac cattgcgctg tccagaggtt cataaagctg
     3121 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     3181 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3241 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3301 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3361 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3421 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3481 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3541 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3601 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3661 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3721 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3781 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3841 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3901 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3961 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4081 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4141 ttagtcagca accatagtcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4261 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4321 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4381 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4441 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4501 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4561 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4621 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4681 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4741 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4801 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4861 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4921 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4981 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5041 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5101 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5161 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5221 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5281 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5341 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5401 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5461 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5521 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5581 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5881 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6421 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcat
//
