LOCUS       sequence-177631-        6990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express human Nup88 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  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 6990)
  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
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             109..903
                     /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             109..903
                     /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(163..182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     773..792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1091..1110)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1135..1182
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1511..2099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2000..2019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2274..2577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2578..2781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2732..2752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2820..2829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2826..2828,2829..2831,2832..3542)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2826..2828,2829..2831,2832..3542)
                     /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(2871..2892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3132..3151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3479..3500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5991..6112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6028..6047)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6082..6101
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6119..6574)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6256..6277)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6468..6487
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6601..6705
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6707..6990,1..74)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6707..6898
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6730..6750)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6915..6990,1..60)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
       61 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
      301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
      661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
      961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     1081 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1141 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     1201 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     1261 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     1321 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     1381 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1621 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2101 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2161 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     2221 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2281 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2341 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2401 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2461 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2521 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2581 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2641 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2701 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2761 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
     2821 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2881 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2941 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3001 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3061 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3121 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3181 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3241 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3301 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3361 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3421 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3481 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3541 agtccggact cagatctcga cttcccgggg cggccgccga gggaccggtg ggcgacggcg
     3601 agctgtggca gacctggctt cctaaccacg tcgtgttctt gcggctccgg gagggactga
     3661 aaaaccagag tccaaccgaa gctgagaaac cagcttcttc gtcgttgcct tcgtcgccgc
     3721 cgccgcagtt gctgacgaga aacgtggtct ttggcctcgg cggagagctt ttcctgtggg
     3781 acggagaaga cagctccttc ttagtcgttc gccttcgggg ccccagcggc ggcggcgaag
     3841 agcccgccct gtcccagtac cagagattgc tttgcataaa tccacccctg tttgaaatct
     3901 atcaagtctt gttaagccca acacaacatc atgtagcact tataggaata aaaggactta
     3961 tggtattaga attacctaaa agatggggga agaattctga atttgaaggt ggaaaatcaa
     4021 cagtgaattg tagtaccact ccagttgcgg agagattttt caccagttcc acctctctga
     4081 ctctaaagca tgctgcatgg tatccaagtg aaatcctgga tccccacgta gtgctgttaa
     4141 catcagacaa cgtaatcaga atttactcac tacgtgagcc gcagacaccc actaacgtga
     4201 taatactttc agaagccgaa gaggaaagtc tagtactcaa taaaggaagg gcgtataccg
     4261 catctctagg agagacagca gttgcatttg actttgggcc attggcagca gtcccaaaga
     4321 ctctatttgg acaaaacggc aaagatgaag tagtggcata cccactgtac atcttatatg
     4381 aaaatggaga gactttcctg acatacatca gtctgttaca cagccctgga aatattggaa
     4441 agctgttggg tccattgccc atgcatcctg cggctgaaga taactatggt tatgatgcgt
     4501 gtgctgtact ctgcttaccc tgtgtcccca atatcttagt gatcgctact gaatcaggaa
     4561 tgctgtatca ctgtgtcgtg ctagaagggg aagaagaaga tgaccacacg tcagaaaagt
     4621 cctgggattc caggattgac ctcattcctt ctctgtatgt gtttgaatgt gttgagttgg
     4681 agcttgcttt gaaactggca tctggagagg atgacccttt tgattctgac ttttcttgtc
     4741 cagtcaaact tcatagagat cccaagtgtc cttcaagata tcactgtact catgaagctg
     4801 gtgtacatag tgttgggcta acttggattc ataaacttca caaatttctt ggatcagatg
     4861 aagaagataa ggatagttta caggaactct ctacagaaca gaaatgcttt gttgaacaca
     4921 tcctttgtac gaagccattg ccctgcaggc agccagctcc aattcgagga ttttggattg
     4981 tacctgacat tctgggaccc acgatgatct gcatcaccag tacctatgaa tgcctcatat
     5041 ggccgttatt aagtacagtc catccagcgt ctcctcccct gctttgtact cgagaagatg
     5101 ttgaagtggc agagtctccc ctccgtgttc tggctgaaac cccagattcc tttgaaaagc
     5161 atattagaag cattttgcaa cgtagtgttg ccaatccagc atttttgaaa gcttctgaaa
     5221 aggacatagc ccctcctcct gaagaatgcc ttcagctcct cagcagagcc acccaggtgt
     5281 tcagagagca gtacattctc aaacaggact tggcaaagga ggagattcag cggagggtca
     5341 aattattatg tgaccaaaaa aagaaacaac tagaagatct cagttattgt cgagaagaga
     5401 ggaaaagtct gcgggaaatg gctgagcgtt tagctgacaa atatgaggaa gctaaagaaa
     5461 aacaagagga tatcatgaac aggatgaaaa aactacttca cagttttcac tctgagctcc
     5521 cagttctctc tgatagtgag cgagacatga agaaagaatt acagctgata cctgatcaac
     5581 ttcgacattt gggcaatgcc atcaaacagg ttactatgaa aaaggattat caacagcaaa
     5641 agatggagaa ggtgttgagt cttccaaaac ccaccattat tctcagtgcc taccagcgaa
     5701 agtgcattca gtccatcctg aaagaggagg gtgaacatat aagggaaatg gtgaagcaaa
     5761 tcaatgatat ccgcaatcat gtaaacttct gacaccacca ggagctgact cacacctgaa
     5821 ctgaacacca ttgtctagac tcgaggatcc gagctccgcg gagatccacc ggatctagat
     5881 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     5941 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     6001 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6061 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     6121 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     6181 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     6241 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     6301 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     6361 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6421 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6481 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6541 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     6601 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6661 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     6721 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     6781 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     6841 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     6901 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6961 gctgactaat tttttttatt tatgcagagg
//
