LOCUS       sequence-177634-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express human Nup35 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  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 5701)
  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-Nup35
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    128..175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      504..1092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     993..1012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1267..1570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1571..1774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1725..1745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1813..1822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1819..1821,1822..1824,1825..2535)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1819..1821,1822..1824,1825..2535)
                     /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(1864..1885)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2125..2144)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2472..2493
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3695..3816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3732..3751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3786..3805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3823..4278)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3960..3981)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4172..4191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4305..4409
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4411..4768
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4411..4602
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4434..4454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4619..4754
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4803..5597
                     /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             4803..5597
                     /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(4857..4876)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5467..5486
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
       61 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
      121 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
      181 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
      241 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
      301 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
      361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
      481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1141 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1201 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1261 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1321 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1381 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1441 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1501 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1561 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     1621 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     1681 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     1741 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     1801 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1861 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1921 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1981 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2041 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2101 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2161 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2221 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2281 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2341 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2401 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2461 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2521 gacgagctgt acaagtccgg actcagatct cgagacgcag cctttgcagt ggaacctcag
     2581 gggcccgcgt taggatctga accaatgatg ctgggttcac ccacatctcc aaagccagga
     2641 gttaatgccc agttcttacc tggattttta atgggggatt tgccagctcc ggtgactcca
     2701 caacctcgat caattagtgg cccttcagta ggagtaatgg aaatgagatc acctttactt
     2761 gcaggtgggt caccaccaca accagttgta ccagctcata aagataaaag tggcgctcca
     2821 ccagttagaa gtatatatga tgacatttct agcccaggac ttggatcaac acctttaact
     2881 tcaagaagac agccaaacat ttcagtaatg cagagtcctc ttgttggagt tacatctact
     2941 cctggaacag ggcaaagtat gtttagtcca gcaagtatcg gtcagccacg aaagacgaca
     3001 ttatctcctg cccagttgga tcctttttat actcaaggag attctttgac ttcagaagat
     3061 cacctcgatg actcttgggt gactgtattt gggtttcctc aagcatctgc ttcctacata
     3121 ttactacaat ttgcacagta tgggaatatc ttaaaacatg tgatgtctaa tacaggaaat
     3181 tggatgcata ttcgttatca atctaaactg caggctcgga aagccttaag caaagatggg
     3241 aggatttttg gagaatccat catgattggt gtaaaaccat gtattgacaa aagtgttatg
     3301 gaaagcagtg acagatgtgc tttatcatct ccatctttag cctttacacc accaatcaaa
     3361 actctaggta caccaacaca acctggaagt actcctagga tttctaccat gagacctctt
     3421 gctacagcat acaaagcctc tactagtgat tatcaggtta tttctgacag acaaacgcca
     3481 aaaaaagatg aaagtcttgt atccaaagca atggagtaca tgtttggctg gtaggaattc
     3541 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     3601 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3661 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3721 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3781 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3841 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3901 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3961 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4021 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4081 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4141 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4201 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4261 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4381 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4441 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4501 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4561 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4621 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4681 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4741 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4801 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4861 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4921 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4981 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5041 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5101 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5161 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5221 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5281 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5341 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5401 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5461 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5521 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5581 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5641 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5701 c
//
