LOCUS       sequence-177632-        8172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express human Nup133 in mammalian cells. It contains synonymous 
            mutations that are not targeted by shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8172)
  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 8172)
  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-mut
FEATURES             Location/Qualifiers
     source          1..8172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             44..838
                     /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             44..838
                     /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(98..117)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     708..727
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1026..1045)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1070..1117
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1446..2034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1935..1954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2209..2512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2513..2716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2667..2687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2755..2764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2761..2763,2764..2766,2767..3477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2761..2763,2764..2766,2767..3477)
                     /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(2806..2827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3067..3086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3414..3435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7108..7229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7145..7164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7199..7218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7236..7691)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7373..7394)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7585..7604
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7718..7822
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(7824..8172,1..9)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7824..8015
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7847..7867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      8032..8167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
       61 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      121 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
      181 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
      241 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
      301 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
      361 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
      421 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
      481 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
      541 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
      601 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
      661 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
      721 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
      781 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
      841 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
      901 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
      961 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     1021 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     1081 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     1141 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     1201 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     1261 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     1321 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1381 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2101 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     2161 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2221 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2281 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2341 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2401 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2461 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     2521 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     2581 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2641 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2701 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
     2761 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2821 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2881 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2941 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3001 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3061 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3121 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3181 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3241 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3301 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3361 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3421 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
     3481 ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtacccc gcggaccccg
     3541 ggtaccgggt cccgaagggg cccgctggcc ggactcgggc ccggctccac gccccggacg
     3601 gctagcagga agggtctgcc cctggggtct gcagtcagct ccccagtgct cttctcgccg
     3661 gtcggccggc gtagctcgct aagctcgcgg ggaacaccaa cacgaatgtt cccacaccac
     3721 tccataactg agtctgtgaa ctatgatgtg aaaacgtttg gatcttctct tcctgttaaa
     3781 gtcatggaag ccctaacatt ggctgaagtc gatgaccagc tgaccattaa catagatgaa
     3841 ggtggatggg cttgtctggt gtgcaaagag aagctcatta tttggaagat tgctctgtca
     3901 cctattacta agttatccgt ttgcaaagaa cttcagctgc cacctagtga tttccactgg
     3961 agtgccgact tagtggctct ttcttactct tctccctcag gtgaagcaca ttctactcag
     4021 gctgttgctg tcatggttgc caccagagaa ggatctatcc gctattggcc aagccttgct
     4081 ggtgaagata cctacacaga ggcttttgta gattcgggag gtgataagac ttacagtttc
     4141 ctaacagcag tgcagggagg aagttttatt ttgtcttcat caggaagcca actaattcgg
     4201 ttgatacctg agagctcagg aaagattcat cagcatatcc tgcctcaggg gcaaggcatg
     4261 ctttcaggaa ttggtcgaaa agtttcttct ctttttggaa ttttatctcc tagtagtgat
     4321 ctcacacttt caagtgttct ctgggataga gagagatcaa gcttttatag cctgacgagt
     4381 tcaaacatca gtaaatggga attagatgat tcttcagaaa agcatgcata cagttgggat
     4441 ataaatagag ccctgaagga aaacattacc gatgctattt ggggatctga aagtaactat
     4501 gaagctatta aagaaggagt caacattcga tatttggact ttaagcaaaa ctgtgatggg
     4561 ctggtgattt tggcagcagc atggcactca gcagacaatc catgtctcat ctattactct
     4621 ctgataacaa tagaagataa tggttgccaa atgtcagatg cagttactgt agaagtcact
     4681 caatataatc caccttttca gtctgaagac ctgattttgt gtcagttgac ggtcccaaac
     4741 ttttcaaacc agactgccta tctgtataac gaaagtgctg tctatgtgtg ctccacagga
     4801 actgggaaat tttctcttcc ccaggagaaa attgtcttta atgcacaagg agatagtgtt
     4861 ttaggtgctg gtgcctgtgg tggtgttcct atcatttttt ctagaaacag tggactggtg
     4921 tctattactt caagggaaaa tgtgtctata ttggcagaag acttggaagg gtctttagca
     4981 tcttcagttg ctggaccaaa cagtgagagt atgatttttg agaccactac aaagaatgaa
     5041 actatagccc aggaagataa aatcaagttg ctgaaagctg cctttctgca atactgcaga
     5101 aaagatttag gtcatgctca aatggtggtt gatgagctct tttcctctca ctctgatttg
     5161 gattctgatt ctgaactaga cagggcagtt acccaaatca gtgtagacct gatggatgac
     5221 tacccagcat ctgacccacg gtgggctgag tctgtccctg aggaagcacc tgggttcagc
     5281 aatacgtcac tgattatcct tcaccagcta gaagacaaga tgaaagctca ctcttttctt
     5341 atggacttta ttcatcaagt tggcttattt ggacgtctag gcagttttcc agttagaggg
     5401 acaccgatgg ccactcgact gttgctctgt gagcatgccg aaaagctgtc agccgccatt
     5461 gttctcaaga accaccactc ccggctttct gaccttgtca acacagccat attgattgct
     5521 ttgaacaaga gggagtatga aatcccatcc aacctgactc ctgcagatgt ctttttcagg
     5581 gaggtatccc aagtagatac catctgtgag tgcttactgg agcatgagga gcaagtcttg
     5641 agggatgcac ctatggattc cattgaatgg gctgaagtgg tgatcaatgt gaacaatatt
     5701 ctcaaggata tgctgcaggc tgctagtcat tatcgccaaa atagaaactc tttgtataga
     5761 agagaagaat cactagaaaa agaacctgaa tatgttccat ggacggcaac aagtggtcct
     5821 ggtggcatcc gaacggtaat aatacgccag catgagattg tcctgaaggt ggcttatcca
     5881 caggcagaca gcaacctccg aaacatcgtg accgagcagc tggtagccct gatcgattgc
     5941 ttcctggatg gttatgtttc tcagcttaag tctgtggata aatccagtaa tcgggaaaga
     6001 tatgacaatc tggagatgga atacctacag aaaagatcag atctcttatc tcctcttctt
     6061 tcactaggcc agtacctgtg ggctgcttct ctagcagaga aatactgtga ctttgatata
     6121 ttggtacaaa tgtgtgagca gactgacaac cagagccgac tccagcgcta catgacccag
     6181 tttgctgatc agaatttttc agactttctc ttccgttggt atctggagaa aggaaagcga
     6241 ggcaaattat tatctcagcc catttctcag catggacagt tggcaaattt tttgcaagct
     6301 catgaacatc tcagctggtt acatgaaatt aatagccaag aattagaaaa ggctcatgca
     6361 acacttctgg gtttggcaaa tatggaaact cgttactttg caaagaagaa aacccttctt
     6421 ggcttgagta aattggctgc attagcttca gacttttcag aggatatgct acaagaaaaa
     6481 attgaagaaa tggctgagca ggagcgcttt ctactgcatc aggagaccct acctgaacag
     6541 ctgctggcgg agaaacagct aaatctcagt gcgatgccag tattgactgc accacaactc
     6601 attggtctat atatctgtga agaaaataga agagctaatg aatatgattt caagaaagct
     6661 ttggacttgt tggaatatat tgatgaggaa gaagatataa atataaatga tctaaaactg
     6721 gaaatccttt gcaaagctct tcagagagat aactggtcca gttctgatgg caaagatgat
     6781 ccaattgaag tatctaaaga cagtatattt gtgaagatct tacagaaact tttaaaagat
     6841 ggcattcagc tcagtgagta cttgccagaa gtcaaggatt tattgcaagc ggatcagctt
     6901 ggaagcttaa agtccaatcc ttacttcgag tttgttttga aagcaaatta tgaatattac
     6961 gtccaaggtc agatttgagg atccaccgga tctagataac tgatcataat cagccatacc
     7021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     7081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     7141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7201 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     7261 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7321 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     7381 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     7441 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     7501 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     7561 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     7621 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     7681 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7801 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     7861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     7921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     7981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     8041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     8101 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     8161 ggaggcctag gc
//
