LOCUS       sequence-456236-        3553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoswitchable fluorescent protein for correlative light-electron 
            microscopy; mEosEM-E for prokaryotic expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3553)
  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 3553)
  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: pRSET A-mEosEM-E
FEATURES             Location/Qualifiers
     source          1..3553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      369..957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     858..877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1111..1128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1213..1232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1263..1285
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1305..1322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1326..1358
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1362..1370,1371..1385)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1401..2084
                     /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"
     primer_bind     complement(2147..2165)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2161..2208
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2305..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2392..2411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2602..2623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2786..2890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2891..2959,2960..3553,1..198)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3109..3128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
       61 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      481 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1081 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1141 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggatctcg
     1201 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
     1261 attttgttta actttaagaa ggagatatac atatgcgggg ttctcatcat catcatcatc
     1321 atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg
     1381 ataaggatcg atggggatcc atggtgagtg cgattaagcc agacatgagg atcaaactcc
     1441 gtatggaagg caacgtaaac gggcaccact ttgtgatcga cggagaaggt acaggcaagc
     1501 cttatgaggg aaaacagacc atggatcttg aagtcaaaga gggcggacct ctgccttttg
     1561 cctttgatat cctgaccact gcattcgaat acggcaacag ggtattcgtg aaatatccag
     1621 acaacataca agactatttt aagcagtcgt ttcctaaggg gtattcgtgg gaacgaagca
     1681 tgactttcga agacgggggc atttgcaatg ccagaaacga cataacaatg gaaggggaca
     1741 ctttctataa taaagttcga ttttatggta ccaactttcc cgccaatggt ccagttatgc
     1801 agaagaagac gctgaaatgg gagccctcca ctgagaaaat gtatgtgcgt gatggagtgc
     1861 tgacgggtga tattgagatg gctttgttgc ttgaaggagg tgcccattac cgatgtgact
     1921 tcagaactac ttacaaagct aaggagaagg gtgtcaagtt accaggcgcc cactttgtgg
     1981 accacgccat tgagatttta agccatgaca aagattacaa caaggttaag ctgtatgagc
     2041 atgctgttgc tcattctgga ttgcctgaca atgccagacg ataagaattc gaagcttgat
     2101 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2161 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     2221 actatatccg gatctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2281 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2341 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2401 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2461 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2521 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2581 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2641 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2701 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2761 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2821 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2881 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2941 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3001 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3061 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3121 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3181 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3241 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3301 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3361 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3421 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3481 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3541 acttctgcgc tcg
//
