LOCUS       sequence-198910-        8190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Nup133-Ypet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8190)
  AUTHORS   Finan K, Raulf A, Heilemann M
  TITLE     A set of homo-oligomeric standards allows accurate protein counting.
  JOURNAL   Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12049-52. doi: 
            10.1002/anie.201505664. Epub 2015 Aug 20.
  PUBMED    26289028
REFERENCE   2  (bases 1 to 8190)
  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: 
            "10.1002/anie.201505664"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2015-10-5- 5"; volume: "54"; issue: "41"; pages: "12049-52"
COMMENT     Sequence Label: pNup133-Ypet
FEATURES             Location/Qualifiers
     source          1..8190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     495..514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        769..1072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1073..1276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1227..1247
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(4804..4806,4807..4809,4810..5523)
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     primer_bind     complement(5110..5129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    5686..5807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5723..5742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5777..5796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5814..6269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5951..5972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6163..6182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6296..6400
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6402..6759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6402..6593
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6425..6445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6610..6745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6794..7588
                     /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             6794..7588
                     /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(6848..6867)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7458..7477
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7776..7795)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7820..7867
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
       61 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      661 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
      721 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      781 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      841 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      901 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      961 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1021 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1081 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1141 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1201 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     1261 ctatataagc agagctggtt tagtgaaccg tcagatccgc caccggtcgc caccatgttc
     1321 ccagccgccc cttctccgcg gaccccgggt accgggtccc gaaggggccc gctggccgga
     1381 ctcgggcccg gctccacgcc ccggacggct agcaggaagg gtctgcccct ggggtctgca
     1441 gtcagctccc cagtgctctt ctcgccggtc ggccggcgta gctcgctaag ctcgcgggga
     1501 acaccaacac gaatgttccc acaccactcc ataactgagt ctgtgaacta tgatgtgaaa
     1561 acgtttggat cttctcttcc tgttaaagtc atggaagccc taacattggc tgaagtcgat
     1621 gaccagctga ccattaacat agatgaaggt ggatgggctt gtctggtgtg caaagagaag
     1681 ctcattattt ggaagattgc tctgtcacct attactaagt tatccgtttg caaagaactt
     1741 cagctgccac ctagtgattt ccactggagt gccgacttag tggctctttc ttactcttct
     1801 ccctcaggtg aagcacattc tactcaggct gttgctgtca tggttgccac cagagaagga
     1861 tctatccgct attggccaag ccttgctggt gaagatacct acacagaggc ttttgtagat
     1921 tcgggaggtg ataagactta cagtttccta acagcagtgc agggaggaag ttttattttg
     1981 tcttcatcag gaagccaact aattcggttg atacctgaga gctcaggaaa gattcatcag
     2041 catatcctgc ctcaggggca aggcatgctt tcaggaattg gtcgaaaagt ttcttctctt
     2101 tttggaattt tatctcctag tagtgatctc acactttcaa gtgttctctg ggatagagag
     2161 agatcaagct tttatagcct gacgagttca aacatcagta aatgggaatt agatgattct
     2221 tcagaaaagc atgcatacag ttgggatata aatagagccc tgaaggaaaa cattaccgat
     2281 gctatttggg gatctgaaag taactatgaa gctattaaag aaggagtcaa cattcgatat
     2341 ttggacttta agcaaaactg tgatgggctg gtgattttgg cagcagcatg gcactcagca
     2401 gacaatccat gtctcatcta ttactctctg ataacaatag aagataatgg ttgccaaatg
     2461 tcagatgcag ttactgtaga agtcactcaa tataatccac cttttcagtc tgaagacctg
     2521 attttgtgtc agttgacggt cccaaacttt tcaaaccaga ctgcctatct gtataacgaa
     2581 agtgctgtct atgtgtgctc cacaggaact gggaaatttt ctcttcccca ggagaaaatt
     2641 gtctttaatg cacaaggaga tagtgtttta ggtgctggtg cctgtggtgg tgttcctatc
     2701 attttttcta gaaacagtgg actggtgtct attacttcaa gggaaaatgt gtctatattg
     2761 gcagaagact tggaagggtc tttagcatct tcagttgctg gaccaaacag tgagagtatg
     2821 atttttgaga ccactacaaa gaatgaaact atagcccagg aagataaaat caagttgctg
     2881 aaagctgcct ttctgcaata ctgcagaaaa gatttaggtc atgctcaaat ggtggttgat
     2941 gagctctttt cctctcactc tgatttggat tctgattctg aactagacag ggcagttacc
     3001 caaatcagtg tagacctgat ggatgactac ccagcatctg acccacggtg ggctgagtct
     3061 gtccctgagg aagcacctgg gttcagcaat acgtcactga ttatccttca ccagctagaa
     3121 gacaagatga aagctcactc ttttcttatg gactttattc atcaagttgg cttatttgga
     3181 cgtctaggca gttttccagt tagagggaca ccgatggcca ctcgactgtt gctctgtgag
     3241 catgccgaaa agctgtcagc cgccattgtt ctcaagaacc accactcccg gctttctgac
     3301 cttgtcaaca cagccatatt gattgctttg aacaagaggg agtatgaaat cccatccaac
     3361 ctgactcctg cagatgtctt tttcagggag gtatcccaag tagataccat ctgtgagtgc
     3421 ttactggagc atgaggagca agtcttgagg gatgcaccta tggattccat tgaatgggct
     3481 gaagtggtga tcaatgtgaa caatattctc aaggatatgc tgcaggctgc tagtcattat
     3541 cgccaaaata gaaactcttt gtatagaaga gaagaatcac tagaaaaaga acctgaatat
     3601 gttccatgga cggcaacaag tggtcctggt ggcatccgaa cggtaataat acgccagcat
     3661 gagattgtcc tgaaggtggc ttatccacag gcagacagca acctccgaaa catcgtgacc
     3721 gagcagctgg tagccctgat cgattgcttc ctggatggtt atgtttctca gcttaagtct
     3781 gtggataaat ccagtaatcg ggaaagatat gacaatctgg agatggaata cctacagaaa
     3841 agatcagatc tcttatctcc tcttctttca ctaggccagt acctgtgggc tgcttctcta
     3901 gcagagaaat actgtgactt tgatatattg gtacaaatgt gtgagcagac tgacaaccag
     3961 agccgactcc agcgctacat gacccagttt gctgatcaga atttttcaga ctttctcttc
     4021 cgttggtatc tggagaaagg aaagcgaggc aaattattat ctcagcccat ttctcagcat
     4081 ggacagttgg caaatttttt gcaagctcat gaacatctca gctggttaca tgaaattaat
     4141 agccaagaat tagaaaaggc tcatgcaaca cttctgggtt tggcaaatat ggaaactcgt
     4201 tactttgcaa agaagaaaac ccttcttggc ttgagtaaat tggctgcatt agcttcagac
     4261 ttttcagagg atatgctaca agaaaaaatt gaagaaatgg ctgagcagga gcgctttcta
     4321 ctgcatcagg agaccctacc tgaacagctg ctggcggaga aacagctaaa tctcagtgcg
     4381 atgccagtat tgactgcacc acaactcatt ggtctatata tctgtgaaga aaatagaaga
     4441 gctaatgaat atgatttcaa gaaagctttg gacttgttgg aatatattga tgaggaagaa
     4501 gatataaata taaatgatct aaaactggaa atcctttgca aagctcttca gagagataac
     4561 tggtccagtt ctgatggcaa agatgatcca attgaagtat ctaaagacag tatatttgtg
     4621 aagatcttac agaaactttt aaaagatggc attcagctca gtgagtactt accggaggtg
     4681 aaagacctgc tacaagcgga tcagcttgga agcttaaagt ccaatcctta cttcgagttt
     4741 gttttgaaag caaattatga atattatgtt cagggacaaa tagctagcgg agcatccgca
     4801 ggaatggtga gcaaaggcga agagctgttc accggcgtgg tgcccatcct ggtggagctg
     4861 gacggcgacg tgaacggcca caagttcagc gtgagcggcg agggcgaggg cgacgccacc
     4921 tacggcaagc tgaccctgaa gctgctgtgc accaccggca agctgcccgt gccctggccc
     4981 accctggtga ccaccctggg ctacggcgtg cagtgcttcg cccggtaccc cgaccacatg
     5041 aagcagcacg acttcttcaa gagcgccatg cccgagggct acgtgcagga gcggaccatc
     5101 ttcttcaagg acgacggcaa ctacaagacc cgggccgagg tgaagttcga gggcgacacc
     5161 ctggtgaacc ggatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggc
     5221 cacaagctgg agtacaacta caacagccac aacgtgtaca tcaccgccga caagcagaag
     5281 aacggcatca aggccaactt caagatccgg cacaacatcg aggacggcgg cgtgcagctg
     5341 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5401 cactacctga gctaccagag cgccctgttc aaggacccca acgagaagcg ggaccacatg
     5461 gtgctgctgg agttcctgac cgccgccggc atcaccgagg gcatgaacga gctctataag
     5521 tagcggccgc atcgataaca ccgcaaatgc ttctagggat ccaccggatc tagataactg
     5581 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     5641 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     5701 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5761 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     5821 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5881 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5941 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6001 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6061 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6121 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6181 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6241 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     6301 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6361 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     6421 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     6481 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     6541 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     6601 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6661 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6721 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     6781 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6841 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6901 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6961 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     7021 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     7081 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     7141 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     7201 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     7261 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     7321 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     7381 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     7441 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     7501 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7561 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     7621 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     7681 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7741 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     7801 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     7861 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     7921 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     7981 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     8041 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     8101 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     8161 gcgtcagacc ccgtagaaaa gatcaaagga
//
