LOCUS       sequence-321886-        6450 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 6450)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6450)
  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..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        411..614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    637..717
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1989..2010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2250..2269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2409..2660
                     /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     2597..2618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3338..3459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3375..3394)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3429..3448
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3466..3921)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3603..3624)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3815..3834
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3948..4052
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4054..4245
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4077..4097)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             4392..5186
                     /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             4392..5186
                     /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(4446..4465)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5056..5075
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5374..5393)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5418..5465
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5794..6382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6283..6302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      661 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtccc
      721 gcgacatggc cacggggttc tcctttgggt ccgggaccct cggctccacc actgtggccc
      781 ccggcggcac cggcacaggc agcggattct ccttcggcgc gtccagcaca ccttctgtgg
      841 ggctcaactt cggaactctc ggaagctctg cgacaccagc atctacatct acttctgcca
      901 gtggttttgg aacaggactc tttggatcta agcctggcac tggattcact ctaggaggaa
      961 caagtgcagg aacaacagct actacatctg catctacaac aggctttagt ttaggattca
     1021 gtaaacctgc agcatctgcc acaccatttg ccctgcccgt cacttctacc acagctagtg
     1081 gtcttactct ctcgtctgct ctgacatcag ctccagcagc atccacagga tttacgttga
     1141 ataacttggg agcaacacca gccacaacta cagcagcatc aacaggactt tccttagggg
     1201 gggccttagc tggtttggga ggttcacttt tccagagtgg aaacacagca acatcaggtc
     1261 tcggacagaa tgctttaagc ttgtctctgg ggactgcaac acctacttca gcagccagca
     1321 gtgaaggcct tggtggtata gattttagta cctcatcaga taaaaagagt gataaaacgg
     1381 gaacaaggcc agaggatagc aaagccctga aagatgaaaa ccttcctcct gtcatctgtc
     1441 aggatgttga gaatctccag aaatttgtca aggaacaaaa acaagtccaa gaagaaatta
     1501 gtcgaatgtc ttccaaagca atgcttaaag tccaggagga catcaaggcc ctgaagcagc
     1561 tcctgtcatt ggctgctagt gggttgcaga ggaacacact caatattgac aaactgaagt
     1621 tagaaactgc tcaggagtta aagaatgcgg aaatagcttt aagaacccag aaaacaccac
     1681 ctggactcca acatgaaaat acagctcctg ctgactactt cagagtcttg gttcagcaat
     1741 ttgaggtaca gcttcagcag tacaggcagc agattgaaga actagaaaac catcttgcca
     1801 ctcaagcaaa caattcacat ataactccgc aagacttatc aatggctatg cagaaaattt
     1861 atcaaacatt tgtagcttta gctgcacaac ttcagtccat tcatgaaaat gtcaaggtgc
     1921 ttaaagaaca gtatctcagc tacagaaaaa tgtttttggg tgagctgttc accggggtgg
     1981 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2041 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2101 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2161 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2221 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2281 tgaagttcga gggcgacaca ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2341 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2401 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2461 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2521 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2581 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2641 gcatggacga gctgtacaag gatgtgtttg aagccaggag aacagaagcc aagaagtggc
     2701 agaatgcacc cagagttact actggaccca cgcctttcag caccatgcca aacgcagcag
     2761 ccgttgccat ggctgcaaca cttacacagc agcaacagcc tgctacaggg ccacagccgt
     2821 ctctgggagt tagttttgga acgccattcg gctcaggtat tggcactggc ttgcaatcaa
     2881 gtggcttagg ttcttcaaac cttggaggat ttggaactag ctctggtttc ggatgcggta
     2941 ccacaggggc ctccacattt ggatttggaa caacagataa accctcagga agtcttagtg
     3001 caggctttgg cagctcaagt acgtctgggt ttaacttcag caatcctggc atcacggcat
     3061 cagctggttt gacatttggg gtgtccaatc ctgcgtctgc ggggtttgga acaggaggac
     3121 aactccttca gttgaagaga cctcccgctg gaaacaaaag aggaaagaga tgaccattgc
     3181 gctgtccaga ggttcataaa gctgtgtaca agtaaagcgg ccgcgactct agatcataat
     3241 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3301 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3361 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3421 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3481 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3541 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3601 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3661 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3721 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3781 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3841 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3901 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3961 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4021 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4081 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4141 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4201 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc atggctgact
     4261 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4321 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4381 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4441 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4501 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4561 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4621 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4681 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4741 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4801 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4861 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4921 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4981 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5041 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5101 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5161 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5221 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5281 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5341 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5401 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5461 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5521 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5581 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5641 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5701 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6421 ttgctggcct tttgctcaca tgttctttcc
//
