LOCUS       sequence-177635-        6949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express human Nup88 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 6949)
  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 6949)
  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-C1X-Nup88-mut
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             136..930
                     /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             136..930
                     /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(190..209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     800..819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1118..1137)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1162..1209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1538..2126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2027..2046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2301..2604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2605..2808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2759..2779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2847..2856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2853..2855,2856..2858,2859..3569)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2853..2855,2856..2858,2859..3569)
                     /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(2898..2919)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3159..3178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3506..3527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5977..6098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6014..6033)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6068..6087
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6105..6560)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6242..6263)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6454..6473
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6587..6691
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6693..6949,1..101)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6693..6884
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6716..6736)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6901..6949,1..87)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
       61 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
      121 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
      181 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
      241 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
      301 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
      361 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
      421 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
      481 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
      541 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
      601 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
      661 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
      721 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
      781 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
      841 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
      901 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
      961 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     1021 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1081 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     1141 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     1201 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     1261 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     1321 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     1381 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1441 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1501 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1561 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1621 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1681 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1741 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1801 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1861 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1921 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1981 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2041 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2101 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2161 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2221 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     2281 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     2341 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2401 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2461 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2521 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2581 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2641 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2701 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2761 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
     2821 cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
     2881 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2941 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3001 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     3061 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3121 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3181 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3241 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3301 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3361 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3421 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     3481 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3541 tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgactt cccggggcgg
     3601 ccgccgaggg accggtgggc gacggcgagc tgtggcagac ctggcttcct aaccacgtcg
     3661 tgttcttgcg gctccgggag ggactgaaaa accagagtcc aaccgaagct gagaaaccag
     3721 cttcttcgtc gttgccttcg tcgccgccgc cgcagttgct gacgagaaac gtggtctttg
     3781 gcctcggcgg agagcttttc ctgtgggacg gagaagacag ctccttctta gtcgttcgcc
     3841 ttcggggccc cagcggcggc ggcgaagagc ccgccctgtc ccagtaccag agattgcttt
     3901 gcataaatcc acccctgttt gaaatctatc aagtcttgtt aagcccaaca caacatcatg
     3961 tagcacttat aggaataaaa ggacttatgg tattagaatt acctaaaaga tgggggaaga
     4021 attctgaatt tgaaggtgga aaatcaacag tgaattgtag taccactcca gttgcggaga
     4081 gatttttcac cagttccacc tctctgactc taaagcatgc tgcatggtat ccaagtgaaa
     4141 tcctggatcc ccacgtagtg ctgttaacat cagacaacgt aatcagaatt tactcactac
     4201 gtgagccgca gacacccact aacgtgataa tactttcaga agccgaagag gaaagtctag
     4261 tactcaataa aggaagggcg tataccgcat ctctaggaga gacagcagtt gcatttgact
     4321 ttgggccatt ggcagcagtc ccaaagactc tatttggaca aaacggcaaa gatgaagtag
     4381 tggcataccc actgtacatc ttatatgaaa atggagagac tttcctgaca tacatcagtc
     4441 tgttacacag ccctggaaat attggaaagc tgttgggtcc attgcccatg catcctgcgg
     4501 ctgaagataa ctatggttat gatgcgtgtg ctgtactctg cttaccctgt gtccccaata
     4561 tcttagtgat cgctactgaa tcaggaatgc tgtatcactg tgtcgtgcta gaaggggaag
     4621 aagaagatga ccacacgtca gaaaagtcct gggattccag gattgacctc attccttctc
     4681 tgtatgtgtt tgaatgtgtt gagttggagc ttgctttgaa actggcatct ggagaggatg
     4741 acccttttga ttctgacttt tcttgtccag tcaaacttca tagagatccc aagtgtcctt
     4801 caagatatca ctgtactcat gaagctggtg tacatagtgt tgggctaact tggattcata
     4861 aacttcacaa atttcttgga tcagatgaag aagataagga tagtttacag gaactctcta
     4921 cagaacagaa atgctttgtt gaacacatcc tttgtacgaa gccattgccc tgcaggcagc
     4981 cagctccaat tcgaggattt tggattgtac ctgacattct gggacccacg atgatctgca
     5041 tcaccagtac ctatgaatgc ctcatatggc cgttattaag tacagtccat ccagcgtctc
     5101 ctcccctgct ttgtactcga gaagatgttg aagtggcaga gtctcccctc cgtgttctgg
     5161 ctgaaacccc agattccttt gaaaagcata ttagaagcat tttgcaacgt agtgttgcca
     5221 atccagcatt tttgaaagct tcagagaaag atattgctcc tcctcctgaa gaatgccttc
     5281 agctcctcag cagagccacc caggtgttca gagagcagta cattctcaaa caggacttgg
     5341 caaaggagga gattcagcgg agggtcaaat tattatgtga ccaaaaaaag aaacaactag
     5401 aagatctcag ttattgtcga gaagagagga aaagtctgcg ggaaatggct gagcgtttag
     5461 ctgacaaata tgaggaagct aaagaaaaac aagaggatat catgaacagg atgaaaaaac
     5521 tacttcacag ttttcactct gagctcccag ttctctctga tagtgagcga gacatgaaga
     5581 aagaattaca gctgatacct gatcaacttc gacatttggg caatgccatc aaacaggtta
     5641 ctatgaaaaa ggattatcaa cagcaaaaga tggagaaggt gttgagtctt ccaaaaccca
     5701 ccattattct cagtgcctac cagcgaaagt gcattcagtc catcctgaaa gaggagggtg
     5761 aacatataag ggaaatggtg aagcaaatca atgatatccg taaccacgtc aatttttagt
     5821 ctagactcga ggatccgagc tccgcggaga tccaccggat ctagataact gatcataatc
     5881 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     5941 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     6001 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6061 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     6121 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     6181 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     6241 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     6301 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     6361 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6421 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6481 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6541 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6601 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6661 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     6721 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6781 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6841 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6901 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggct
//
