LOCUS       sequence-177626-        8368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express human Nup133 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8368)
  AUTHORS   Tie HC, Madugula V, Lu L
  TITLE     The development of a single molecule fluorescence standard and its 
            application in estimating the stoichiometry of the nuclear pore 
            complex.
  JOURNAL   Biochem Biophys Res Commun. 2016 Sep 30;478(4):1694-9. doi: 
            10.1016/j.bbrc.2016.09.005. Epub 2016 Sep 6.
  PUBMED    27613095
REFERENCE   2  (bases 1 to 8368)
  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"; doi: 
            "10.1016/j.bbrc.2016.09.005"; journalName: "Biochem Biophys Res 
            Commun"; date: "2016-09-30- 30"; volume: "478"; issue: "4"; pages: 
            "1694-9"
COMMENT     Sequence Label: pEGFP-C1-KSENB-Nup133
FEATURES             Location/Qualifiers
     source          1..8368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      573..582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(579..581,582..584,585..1295)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(579..581,582..584,585..1295)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(624..645)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(885..904)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1232..1253
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5122..5243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5159..5178)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5213..5232
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5250..5705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5387..5408)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5599..5618
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5732..5836
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5838..6195
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5838..6029
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5861..5881)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6046..6181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6230..7024
                     /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             6230..7024
                     /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(6284..6303)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6894..6913
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7212..7231)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7256..7303
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7632..8220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8121..8140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      541 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      601 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      661 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      721 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      781 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      841 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      901 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      961 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1021 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1081 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1141 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1201 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1261 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag
     1321 cttcgaattc tgcagtcgac ggtaccccgc ggaccccggg taccgggtcc cgaaggggcc
     1381 cgctggccgg actcgggccc ggctccacgc cccggacggc tagcaggaag ggtctgcccc
     1441 tggggtctgc agtcagctcc ccagtgctct tctcgccggt cggccggcgt agctcgctaa
     1501 gctcgcgggg aacaccaaca cgaatgttcc cacaccactc cataactgag tctgtgaact
     1561 atgatgtgaa aacgtttgga tcttctcttc ctgttaaagt catggaagcc ctaacattgg
     1621 ctgaagtcga tgaccagctg accattaaca tagatgaagg tggatgggct tgtctggtgt
     1681 gcaaagagaa gctcattatt tggaagattg ctctgtcacc tattactaag ttatccgttt
     1741 gcaaagaact tcagctgcca cctagtgatt tccactggag tgccgactta gtggctcttt
     1801 cttactcttc tccctcaggt gaagcacatt ctactcaggc tgttgctgtc atggttgcca
     1861 ccagagaagg atctatccgc tattggccaa gccttgctgg tgaagatacc tacacagagg
     1921 cttttgtaga ttcgggaggt gataagactt acagtttcct aacagcagtg cagggaggaa
     1981 gttttatttt gtcttcatca ggaagccaac taattcggtt gatacctgag agctcaggaa
     2041 agattcatca gcatatcctg cctcaggggc aaggcatgct ttcaggaatt ggtcgaaaag
     2101 tttcttctct ttttggaatt ttatctccta gtagtgatct cacactttca agtgttctct
     2161 gggatagaga gagatcaagc ttttatagcc tgacgagttc aaacatcagt aaatgggaat
     2221 tagatgattc ttcagaaaag catgcataca gttgggatat aaatagagcc ctgaaggaaa
     2281 acattaccga tgctatttgg ggatctgaaa gtaactatga agctattaaa gaaggagtca
     2341 acattcgata tttggacttt aagcaaaact gtgatgggct ggtgattttg gcagcagcat
     2401 ggcactcagc agacaatcca tgtctcatct attactctct gataacaata gaagataatg
     2461 gttgccaaat gtcagatgca gttactgtag aagtcactca atataatcca ccttttcagt
     2521 ctgaagacct gattttgtgt cagttgacgg tcccaaactt ttcaaaccag actgcctatc
     2581 tgtataacga aagtgctgtc tatgtgtgct ccacaggaac tgggaaattt tctcttcccc
     2641 aggagaaaat tgtctttaat gcacaaggag atagtgtttt aggtgctggt gcctgtggtg
     2701 gtgttcctat cattttttct agaaacagtg gactggtgtc tattacttca agggaaaatg
     2761 tgtctatatt ggcagaagac ttggaagggt ctttagcatc ttcagttgct ggaccaaaca
     2821 gtgagagtat gatttttgag accactacaa agaatgaaac tatagcccag gaagataaaa
     2881 tcaagttgct gaaagctgcc tttctgcaat actgcagaaa agatttaggt catgctcaaa
     2941 tggtggttga tgagctcttt tcctctcact ctgatttgga ttctgattct gaactagaca
     3001 gggcagttac ccaaatcagt gtagacctga tggatgacta cccagcatct gacccacggt
     3061 gggctgagtc tgtccctgag gaagcacctg ggttcagcaa tacgtcactg attatccttc
     3121 accagctaga agacaagatg aaagctcact cttttcttat ggactttatt catcaagttg
     3181 gcttatttgg acgtctaggc agttttccag ttagagggac accgatggcc actcgactgt
     3241 tgctctgtga gcatgccgaa aagctgtcag ccgccattgt tctcaagaac caccactccc
     3301 ggctttctga ccttgtcaac acagccatat tgattgcttt gaacaagagg gagtatgaaa
     3361 tcccatccaa cctgactcct gcagatgtct ttttcaggga ggtatcccaa gtagatacca
     3421 tctgtgagtg cttactggag catgaggagc aagtcttgag ggatgcacct atggattcca
     3481 ttgaatgggc tgaagtggtg atcaatgtga acaatattct caaggatatg ctgcaggctg
     3541 ctagtcatta tcgccaaaat agaaactctt tgtatagaag agaagaatca ctagaaaaag
     3601 aacctgaata tgttccatgg acggcaacaa gtggtcctgg tggcatccga acggtaataa
     3661 tacgccagca tgagattgtc ctgaaggtgg cttatccaca ggcagacagc aacctccgaa
     3721 acatcgtgac cgagcagctg gtagccctga tcgattgctt cctggatggt tatgtttctc
     3781 agcttaagtc tgtggataaa tccagtaatc gggaaagata tgacaatctg gagatggaat
     3841 acctacagaa aagatcagat ctcttatctc ctcttctttc actaggccag tacctgtggg
     3901 ctgcttctct agcagagaaa tactgtgact ttgatatatt ggtacaaatg tgtgagcaga
     3961 ctgacaacca gagccgactc cagcgctaca tgacccagtt tgctgatcag aatttttcag
     4021 actttctctt ccgttggtat ctggagaaag gaaagcgagg caaattatta tctcagccca
     4081 tttctcagca tggacagttg gcaaattttt tgcaagctca tgaacatctc agctggttac
     4141 atgaaattaa tagccaagaa ttagaaaagg ctcatgcaac acttctgggt ttggcaaata
     4201 tggaaactcg ttactttgca aagaagaaaa cccttcttgg cttgagtaaa ttggctgcat
     4261 tagcttcaga cttttcagag gatatgctac aagaaaaaat tgaagaaatg gctgagcagg
     4321 agcgctttct actgcatcag gagaccctac ctgaacagct gctggcggag aaacagctaa
     4381 atctcagtgc gatgccagta ttgactgcac cacaactcat tggtctatat atctgtgaag
     4441 aaaatagaag agctaatgaa tatgatttca agaaagcttt ggacttgttg gaatatattg
     4501 atgaggaaga agatataaat ataaatgatc taaaactgga aatcctttgc aaagctcttc
     4561 agagagataa ctggtccagt tctgatggca aagatgatcc aattgaagta tctaaagaca
     4621 gtatatttgt gaagatctta cagaaacttt taaaagatgg cattcagctc agtgagtact
     4681 taccggaggt gaaagacctg ctacaagcgg atcagcttgg aagcttaaag tccaatcctt
     4741 acttcgagtt tgttttgaaa gcaaattatg aatattatgt tcagggacaa atataacttt
     4801 ttctaaaaat ggccattgtt tatgaaatct gtataagtgt gtccttatac aaattttagg
     4861 ccataaacaa gtgtaagttt gtacaatttc ataacatgta tagctgagtt tttatacttt
     4921 atatgtagga agctaatata aaatagttat gtaactgtga aaaaaaaaaa aaaaaaaaaa
     4981 aagggcggcc gcggatccac cggatctaga taactgatca taatcagcca taccacattt
     5041 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     5101 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5161 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5221 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     5281 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5341 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5401 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5461 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5521 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5581 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5641 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5701 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5761 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5821 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     5881 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5941 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6001 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6061 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6121 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6181 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6241 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6301 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6361 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6421 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6481 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6541 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6601 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6661 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6721 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     6781 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6841 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6901 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6961 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     7021 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     7081 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     7141 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     7201 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     7261 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     7321 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     7381 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     7441 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     7501 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7561 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7621 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7681 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7741 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7801 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7861 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7921 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7981 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8041 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8101 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8161 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8221 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8281 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     8341 ttaatagtaa tcaattacgg ggtcatta
//
