LOCUS       sequence-321890-        6252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 4 
            amino acids) at the caroxyl-terminus with no linker and a total net 
            fusion of (-4) amino acids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6252)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6252)
  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_mEGFP510(-4)
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    613..693
                     /label=MCS
                     /note="multiple cloning site"
     CDS             join(2223..2225,2226..2939)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VFSGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2265..2286)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2526..2545)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2873..2894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3062..3183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3099..3118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3153..3172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3190..3645)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3327..3348)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3539..3558
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3672..3776
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3778..4135
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3778..3969
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3801..3821)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3986..4121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4170..4964
                     /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             4170..4964
                     /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(4224..4243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4834..4853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5152..5171)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5196..5243
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5572..6160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6061..6080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca
      661 gtcgacggta ccgcgggccc gggatccacc ggtcccgcga catggcattc aattttgggg
      721 ctccttcggg cacttcgggc acctctacag ccaccgcggc accggcgggc gggtttggcg
      781 gatttggaac aacaactaca actgcaggct ctgcattcag cttttctgcc ccaacaaaca
      841 caggcagtac aggccttctc ggcggcactc agaacaaagg ttttggcttt ggcactggtt
      901 ttggcacatc gacgggtact ggcactggtt taggcactgg cttgggaacc ggacttggat
      961 tcggaggatt taacacccag cagcagcagc agcagcagca gacttcttta ggcggtctct
     1021 tcagtcagcc tgcacaggcc cctgcgcagt ccaaccaact catcaacact gccagcgctc
     1081 tttctgctcc aacgctattg ggggatgaga gagatgccat cttggcaaag tggaaccagt
     1141 tgcaggcctt ctggggaaca ggaaaaggat atttcaataa taacattcct ccagtggagt
     1201 ttacacagga aaaccccttt tgccgattta aggctgtcgg ttacagttgt atgcccaata
     1261 acaaagatga agatggacta gtggttctaa ttttcaacaa aaaagaaaca gatattagaa
     1321 gccaacaaca gcagttggta gaatcattgc ataaagttct gggaggaaac cagaccctca
     1381 ctgtcaatgt ggagggcatt aagacactgc ccgacgatca gacagaagtt gtcatttaca
     1441 ttgttgagcg ctctccaaat ggcacctcga gaagagttcc agcgacaacg ctgtatgccc
     1501 actttgagca agccaacatt aaaacacagc ttcagcagct tggcgtgacc ctttctatga
     1561 ccagaacaga gctgtctcct gctcagatca agcagctcct acagaaccct cctgctggtg
     1621 ttgatcccat tatttgggaa caggccaagg tggataaccc ggattctgaa aaattaattc
     1681 ctgttccaat ggtgggtttc aaagaattgc ttcgaagact gaaggttcaa gaccagatga
     1741 ctaagcagca ccagaccaga ttagatatca tatctgaaga tattagtgaa ctgcaaaaga
     1801 atcaaactac aactatggcc aaaatagcac aatacaagag gaaactgatg gatctctcac
     1861 acagaacctt acaggtccta atcaagcagg aaatccagag gaagagtggc tatgccatcc
     1921 aggctgagga agaacagttg cgggttcagc tagatacgat tcaaggagag ctgaatgcac
     1981 ccactcagtt caaaggtcga ctgaatgaac tgatgtccca gatcaggatg cagaatcatt
     2041 ttggagctgt gaaatctgaa gaaaagtatt acatagatgc agacctatta cgagagatca
     2101 agcagcattt gaaacagcaa caggaaggcc ttagccacct gattagcatc ataaaagacg
     2161 acctggaaga tatcaagctg gtagagcatg gactgaatga gaccatccac agcagaggcg
     2221 gggtctttag cggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2281 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2341 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2401 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2461 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2521 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2581 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2641 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2701 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2761 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2821 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2881 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     2941 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3001 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3061 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3121 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3181 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3241 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3301 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3361 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3421 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3481 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3541 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3601 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3661 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3721 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3781 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3841 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4021 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4081 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4141 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4201 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4261 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4321 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4381 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4441 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4501 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4561 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4621 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4681 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4741 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4801 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4861 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4921 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4981 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5041 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5101 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5161 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5221 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5281 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5341 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5401 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5461 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5521 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5581 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5641 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5701 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5761 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5821 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5881 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5941 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6001 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6061 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6121 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6181 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6241 tgattctgtg ga
//
