LOCUS       sequence-321676-        6288 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of Nup54 with a mEGFP (missing the first 6 
            amino acids) at amino acid position 494.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6288)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 6288)
  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(-6)
FEATURES             Location/Qualifiers
     source          1..6288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    634..714
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2232..2253)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2493..2512)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2652..2903
                     /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     2840..2861
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3119..3240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3156..3175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3210..3229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3247..3702)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3384..3405)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3596..3615
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3729..3833
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3835..4192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3835..4026
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3858..3878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4043..4178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4227..5021
                     /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             4227..5021
                     /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(4281..4300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4891..4910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5209..5228)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5253..5300
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5629..6217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6118..6137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
      661 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcccgcg
      721 acatggcatt caattttggg gctccttcgg gcacttcggg cacctctaca gccaccgcgg
      781 caccggcggg cgggtttggc ggatttggaa caacaactac aactgcaggc tctgcattca
      841 gcttttctgc cccaacaaac acaggcagta caggccttct cggcggcact cagaacaaag
      901 gttttggctt tggcactggt tttggcacat cgacgggtac tggcactggt ttaggcactg
      961 gcttgggaac cggacttgga ttcggaggat ttaacaccca gcagcagcag cagcagcagc
     1021 agacttcttt aggcggtctc ttcagtcagc ctgcacaggc ccctgcgcag tccaaccaac
     1081 tcatcaacac tgccagcgct ctttctgctc caacgctatt gggggatgag agagatgcca
     1141 tcttggcaaa gtggaaccag ttgcaggcct tctggggaac aggaaaagga tatttcaata
     1201 ataacattcc tccagtggag tttacacagg aaaacccctt ttgccgattt aaggctgtcg
     1261 gttacagttg tatgcccaat aacaaagatg aagatggact agtggttcta attttcaaca
     1321 aaaaagaaac agatattaga agccaacaac agcagttggt agaatcattg cataaagttc
     1381 tgggaggaaa ccagaccctc actgtcaatg tggagggcat taagacactg cccgacgatc
     1441 agacagaagt tgtcatttac attgttgagc gctctccaaa tggcacctcg agaagagttc
     1501 cagcgacaac gctgtatgcc cactttgagc aagccaacat taaaacacag cttcagcagc
     1561 ttggcgtgac cctttctatg accagaacag agctgtctcc tgctcagatc aagcagctcc
     1621 tacagaaccc tcctgctggt gttgatccca ttatttggga acaggccaag gtggataacc
     1681 cggattctga aaaattaatt cctgttccaa tggtgggttt caaagaattg cttcgaagac
     1741 tgaaggttca agaccagatg actaagcagc accagaccag attagatatc atatctgaag
     1801 atattagtga actgcaaaag aatcaaacta caactatggc caaaatagca caatacaaga
     1861 ggaaactgat ggatctctca cacagaacct tacaggtcct aatcaagcag gaaatccaga
     1921 ggaagagtgg ctatgccatc caggctgagg aagaacagtt gcgggttcag ctagatacga
     1981 ttcaaggaga gctgaatgca cccactcagt tcaaaggtcg actgaatgaa ctgatgtccc
     2041 agatcaggat gcagaatcat tttggagctg tgaaatctga agaaaagtat tacatagatg
     2101 cagacctatt acgagagatc aagcagcatt tgaaacagca acaggaaggc cttagccacc
     2161 tgattagcat cataaaagac gacctggaag atatcaagct ggtagagctg ttcaccgggg
     2221 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2281 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2341 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2401 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2461 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2521 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2581 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2641 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2701 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2761 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     2821 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2881 tcggcatgga cgagctgtac aaggagcatg gactgaatga gaccatccac agcagaggcg
     2941 gggtctttag ctgaccattg cgctgtccag aggttcataa agctgtgtac aagtaaagcg
     3001 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3061 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3241 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3301 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3361 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3421 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3481 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3541 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3601 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3661 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3841 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4021 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4141 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4201 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4261 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4321 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4381 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4441 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4501 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4561 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4621 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4681 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4741 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4801 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4861 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4921 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4981 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5041 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5101 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5161 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5221 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5281 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5341 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5401 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5461 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5521 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgc
//
