LOCUS       sequence-321673-        6312 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 5 
            amino acids) at amino acid position 494 with a flexible linker of 7 
            amino acids at the carboxyl end of the mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6312)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Nup54-mEGFP494(flex2)
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        355..558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     509..529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    581..661
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2203..2224)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2464..2483)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2623..2874
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     2811..2832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3090..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3127..3146)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3181..3200
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3218..3673)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3355..3376)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3567..3586
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3700..3804
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3806..4163
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3806..3997
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3829..3849)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4014..4149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4198..4992
                     /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             4198..4992
                     /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(4252..4271)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4862..4881
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5180..5199)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5224..5271
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5600..6188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6089..6108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      361 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag
      601 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
      661 tcccgcgaca tggcattcaa ttttggggct ccttcgggca cttcgggcac ctctacagcc
      721 accgcggcac cggcgggcgg gtttggcgga tttggaacaa caactacaac tgcaggctct
      781 gcattcagct tttctgcccc aacaaacaca ggcagtacag gccttctcgg cggcactcag
      841 aacaaaggtt ttggctttgg cactggtttt ggcacatcga cgggtactgg cactggttta
      901 ggcactggct tgggaaccgg acttggattc ggaggattta acacccagca gcagcagcag
      961 cagcagcaga cttctttagg cggtctcttc agtcagcctg cacaggcccc tgcgcagtcc
     1021 aaccaactca tcaacactgc cagcgctctt tctgctccaa cgctattggg ggatgagaga
     1081 gatgccatct tggcaaagtg gaaccagttg caggccttct ggggaacagg aaaaggatat
     1141 ttcaataata acattcctcc agtggagttt acacaggaaa accccttttg ccgatttaag
     1201 gctgtcggtt acagttgtat gcccaataac aaagatgaag atggactagt ggttctaatt
     1261 ttcaacaaaa aagaaacaga tattagaagc caacaacagc agttggtaga atcattgcat
     1321 aaagttctgg gaggaaacca gaccctcact gtcaatgtgg agggcattaa gacactgccc
     1381 gacgatcaga cagaagttgt catttacatt gttgagcgct ctccaaatgg cacctcgaga
     1441 agagttccag cgacaacgct gtatgcccac tttgagcaag ccaacattaa aacacagctt
     1501 cagcagcttg gcgtgaccct ttctatgacc agaacagagc tgtctcctgc tcagatcaag
     1561 cagctcctac agaaccctcc tgctggtgtt gatcccatta tttgggaaca ggccaaggtg
     1621 gataacccgg attctgaaaa attaattcct gttccaatgg tgggtttcaa agaattgctt
     1681 cgaagactga aggttcaaga ccagatgact aagcagcacc agaccagatt agatatcata
     1741 tctgaagata ttagtgaact gcaaaagaat caaactacaa ctatggccaa aatagcacaa
     1801 tacaagagga aactgatgga tctctcacac agaaccttac aggtcctaat caagcaggaa
     1861 atccagagga agagtggcta tgccatccag gctgaggaag aacagttgcg ggttcagcta
     1921 gatacgattc aaggagagct gaatgcaccc actcagttca aaggtcgact gaatgaactg
     1981 atgtcccaga tcaggatgca gaatcatttt ggagctgtga aatctgaaga aaagtattac
     2041 atagatgcag acctattacg agagatcaag cagcatttga aacagcaaca ggaaggcctt
     2101 agccacctga ttagcatcat aaaagacgac ctggaagata tcaagctggt agggggtggc
     2161 ggcagctccg gagaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2221 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2281 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2341 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2401 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2461 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2521 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2581 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2641 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2701 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2761 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2821 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggagcat
     2881 ggactgaatg agaccatcca cagcagaggc ggggtcttta gctgaccatt gcgctgtcca
     2941 gaggttcata aagctgtgta caagtaaagc ggccgcgact ctagatcata atcagccata
     3001 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3061 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3121 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3181 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     3241 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3301 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3361 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3421 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3481 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3541 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3601 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3661 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3721 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3781 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3841 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3961 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4021 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4081 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4141 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4201 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4261 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4321 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4381 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4441 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4501 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4561 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4621 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4681 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4741 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4801 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4861 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4921 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4981 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5041 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5101 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5161 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5221 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5281 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5341 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5401 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5461 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5521 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6301 attagttatt aa
//
