LOCUS       sequence-258715-        3553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  application in Correlative light-electron microscopy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3553)
  AUTHORS   Fu Z, Peng D, Zhang M, Xue F, Zhang R, He W, Xu T, Xu P
  TITLE     mEosEM withstands osmium staining and Epon embedding for 
            super-resolution CLEM.
  JOURNAL   Nat Methods. 2019 Oct 14. pii: 10.1038/s41592-019-0613-6. doi: 
            10.1038/s41592-019-0613-6.
  PUBMED    31611693
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: "Nat 
            Methods. 2019 Oct 14. pii: 10.1038/s41592-019-0613-6. doi: 
            10.1038/s41592-019-0613-6."
COMMENT     Sequence Label: pRSET-mEosEM
FEATURES             Location/Qualifiers
     source          1..3553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /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(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             189..872
                     /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
                     KEGGPLPFAFDILTTAFHYGNRVFVKYPDNIQDYFKQSFPKGYSWERSMTFEDGGICNA
                     RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL
                     LEGGAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     primer_bind     complement(935..953)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      949..996
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1093..1548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1180..1199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1390..1411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1574..1678
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1679..1747,1748..2539)
                     /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(1897..1916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2710..3298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3199..3218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3452..3469
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatcgat
      181 ggggatccat ggtgagtgcg attaagccag acatgaggat caaactccgt atggaaggca
      241 acgtaaacgg gcaccacttt gtgatcgacg gagaaggtac aggcaagcct tatgagggaa
      301 aacagaccat ggatcttgaa gtcaaagagg gcggacctct gccttttgcc tttgatatcc
      361 tgaccactgc attccattac ggcaacaggg tattcgtgaa atatccagac aacatacaag
      421 actattttaa gcagtcgttt cctaaggggt attcgtggga acgaagcatg actttcgaag
      481 acgggggcat ttgcaatgcc agaaacgaca taacaatgga aggggacact ttctataata
      541 aagttcgatt ttatggtacc aactttcccg ccaatggtcc agttatgcag aagaagacgc
      601 tgaaatggga gccctccact gagaaaatgt atgtgcgtga tggagtgctg acgggtgata
      661 ttgagatggc tttgttgctt gaaggaggtg cccattaccg atgtgacttc agaactactt
      721 acaaagctaa ggagaagggt gtcaagttac caggcgccca ctttgtggac cacgccattg
      781 agattttaag ccatgacaaa gattacaaca aggttaagct gtatgagcat gctgttgctc
      841 attctggatt gcctgacaat gccagacgat aagaattcga agcttgatcc ggctgctaac
      901 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
      961 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1021 tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1081 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1141 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1201 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1261 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1321 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1381 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     1441 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1501 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     1561 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1621 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1681 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1741 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1801 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1861 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1921 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1981 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2041 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2101 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2161 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2221 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2281 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2341 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2401 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2461 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2521 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2581 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2641 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2701 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2761 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2821 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2881 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2941 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3001 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3061 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3121 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3181 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3241 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3301 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3361 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3421 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3481 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggatctc
     3541 gatcccgcga aat
//
