LOCUS       sequence-456227-        6690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoswitchable fluorescent protein for correlative light-electron 
            microscopy; nuclear envelope-targeted mEosEM-E for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6690)
  AUTHORS   Peng D, Li N, He W, Drasbek KR, Xu T, Zhang M, Xu P
  TITLE     Improved Fluorescent Proteins for Dual-Colour Post-Embedding CLEM.
  JOURNAL   Cells. 2022 Mar 23;11(7):1077. doi: 10.3390/cells11071077.
  PUBMED    35406640
REFERENCE   2  (bases 1 to 6690)
  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"; journalName: "Cells."; 
            date: "2022-03-23"; volume: "11(7):1077. doi"; pages: " 
            10.3390/cells11071077"
COMMENT     Sequence Label: mEosEM-E_Lamin A
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(320..339)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    364..411
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      740..1328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1229..1248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1503..1806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1807..2010
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2049..2058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2055..2735
                     /codon_start=1
                     /product="bright monomeric photocontrollable fluorescent 
                     protein that withstands Epon embedding and osmium tetroxide
                     treatment (Fu et al., 2020)"
                     /label=mEosEM
                     /note="shows on and off switching in the green form after 
                     Epon embedding"
                     /translation="MVSAIKPDMRIKLRMEGNVNGHHFVIDGEGTGKPYEGKQTMDLEV
                     KEGGPLPFAFDILTTAFEYGNRVFVKYPDNIQDYFKQSFPKGYSWERSMTFEDGGICNA
                     RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL
                     LEGGAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     polyA_signal    4920..5041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4957..4976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5011..5030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5048..5503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5185..5206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5397..5416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5530..5634
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5636..5993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5636..5827
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5659..5679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5844..5979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(6028..6690,1..132)
                     /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             join(6028..6690,1..132)
                     /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(6082..6101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
       61 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
      121 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
      181 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
      241 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
      301 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
      361 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
      421 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
      481 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
      541 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
      601 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
      661 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
      721 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      781 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      841 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      901 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      961 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1021 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1081 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1141 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1201 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1261 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1321 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1381 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     1441 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     1501 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1561 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     1621 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     1681 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1741 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1801 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     1861 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     1921 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     1981 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
     2041 taccggtcgc caccatggtg agtgcgatta agccagacat gaggatcaaa ctccgtatgg
     2101 aaggcaacgt aaacgggcac cactttgtga tcgacggaga aggtacaggc aagccttatg
     2161 agggaaaaca gaccatggat cttgaagtca aagagggcgg acctctgcct tttgcctttg
     2221 atatcctgac cactgcattc gaatacggca acagggtatt cgtgaaatat ccagacaaca
     2281 tacaagacta ttttaagcag tcgtttccta aggggtattc gtgggaacga agcatgactt
     2341 tcgaagacgg gggcatttgc aatgccagaa acgacataac aatggaaggg gacactttct
     2401 ataataaagt tcgattttat ggtaccaact ttcccgccaa tggtccagtt atgcagaaga
     2461 agacgctgaa atgggagccc tccactgaga aaatgtatgt gcgtgatgga gtgctgacgg
     2521 gtgatattga gatggctttg ttgcttgaag gaggtgccca ttaccgatgt gacttcagaa
     2581 ctacttacaa agctaaggag aagggtgtca agttaccagg cgcccacttt gtggaccacg
     2641 ccattgagat tttaagccat gacaaagatt acaacaaggt taagctgtat gagcatgctg
     2701 ttgctcattc tggattgcct gacaatgcca gacgatccgg ccggactcag atctcgagct
     2761 caagcttcga attcatggag accccgtccc agcggcgcgc cacccgcagc ggggcgcagg
     2821 ccagctccac tccgctgtcg cccacccgca tcacccggct gcaggagaag gaggacctgc
     2881 aggagctcaa tgatcgcttg gcggtctaca tcgaccgtgt gcgctcgctg gaaacggaga
     2941 acgcagggct gcgccttcgc atcaccgagt ctgaagaggt ggtcagccgc gaggtgtccg
     3001 gcatcaaggc cgcctacgag gccgagctcg gggatgcccg caagaccctt gactcagtag
     3061 ccaaggagcg cgcccgcctg cagctggagc tgagcaaagt gcgtgaggag tttaaggagc
     3121 tgaaagcgcg caataccaag aaggagggtg acctgatagc tgctcaggct cggctgaagg
     3181 acctggaggc tctgctgaac tccaaggagg ccgcactgag cactgctctc agtgagaagc
     3241 gcacgctgga gggcgagctg catgatctgc ggggccaggt ggccaagctt gaggcagccc
     3301 taggtgaggc caagaagcaa cttcaggatg agatgctgcg gcgggtggat gctgagaaca
     3361 ggctgcagac catgaaggag gaactggact tccagaagaa catctacagt gaggagctgc
     3421 gtgagaccaa gcgccgtcat gagacccgac tggtggagat tgacaatggg aagcagcgtg
     3481 agtttgagag ccggctggcg gatgcgctgc aggaactgcg ggcccagcat gaggaccagg
     3541 tggagcagta taagaaggag ctggagaaga cttattctgc caagctggac aatgccaggc
     3601 agtctgctga gaggaacagc aacctggtgg gggctgccca cgaggagctg cagcagtcgc
     3661 gcatccgcat cgacagcctc tctgcccagc tcagccagct ccagaagcag ctggcagcca
     3721 aggaggcaaa gcttcgagac ctggaggact cactggcccg tgagcgggac accagccggc
     3781 ggctgctggc ggaaaaggag cgggagatgg ccgagatgcg ggcaaggatg cagcagcagc
     3841 tggacgagta ccaggagctt ctggacatca agctggccct ggacatggag atccacgcct
     3901 accgcaagct cttggagggc gaggaggaga ggctacgcct gtcccccagc cctacctcgc
     3961 agcgcagccg tggccgtgct tcctctcact catcccagac acagggtggg ggcagcgtca
     4021 ccaaaaagcg caaactggag tccactgaga gccgcagcag cttctcacag cacgcacgca
     4081 ctagcgggcg cgtggccgtg gaggaggtgg acgaggaggg caagtttgtc cggctgcgca
     4141 acaagtccaa tgaggaccag tccatgggca attggcagat caagcgccag aatggagatg
     4201 atcccttgct gacttaccgg ttcccaccaa agttcaccct gaaggctggg caggtggtga
     4261 cgatctgggc tgcaggagct ggggccaccc acagcccccc taccgacctg gtgtggaagg
     4321 cacagaacac ctggggctgc gggaacagcc tgcgtacggc tctcatcaac tccactgggg
     4381 aagaagtggc catgcgcaag ctggtgcgct cagtgactgt ggttgaggac gacgaggatg
     4441 aggatggaga tgacctgctc catcaccacc acggctccca ctgcagcagc tcgggggacc
     4501 ccgctgagta caacctgcgc tcgcgcaccg tgctgtgcgg gacctgcggg cagcctgccg
     4561 acaaggcatc tgccagcggc tcaggagccc aggtgggcgg acccatctcc tctggctctt
     4621 ctgcctccag tgtcacggtc actcgcagct accgcagtgt ggggggcagt gggggtggca
     4681 gcttcgggga caatctggtc acccgctcct acctcctggg caactccagc ccccgaaccc
     4741 agagccccca gaactgcagc atcatgtaag tcgacggtac cgcgggcccg ggatccaccg
     4801 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     4861 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4981 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5041 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5101 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5161 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5221 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5281 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5341 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5401 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5461 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     5521 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5581 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     5641 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5821 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5881 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5941 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     6001 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     6061 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     6121 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     6181 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     6241 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     6301 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     6361 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     6421 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     6481 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6541 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6601 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6661 ctgggtgtgg cggaccgcta tcaggacata
//
