LOCUS       sequence-259169-        6690 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 6690)
  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 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: "Nat 
            Methods. 2019 Oct 14. pii: 10.1038/s41592-019-0613-6. doi: 
            10.1038/s41592-019-0613-6."
COMMENT     Sequence Label: mEosEM-Lamin A
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             554..1234
                     /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"
     polyA_signal    3419..3540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3456..3475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3510..3529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3547..4002)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3684..3705)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3896..3915
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4029..4133
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4135..4492
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4135..4326
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4158..4178)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4343..4478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4527..5321
                     /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             4527..5321
                     /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(4581..4600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5191..5210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5509..5528)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5553..5600
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5929..6517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6418..6437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggtcgcc accatggtga gtgcgattaa gccagacatg aggatcaaac tccgtatgga
      601 aggcaacgta aacgggcacc actttgtgat cgacggagaa ggtacaggca agccttatga
      661 gggaaaacag accatggatc ttgaagtcaa agagggcgga cctctgcctt ttgcctttga
      721 tatcctgacc actgcattcc attacggcaa cagggtattc gtgaaatatc cagacaacat
      781 acaagactat tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa gcatgacttt
      841 cgaagacggg ggcatttgca atgccagaaa cgacataaca atggaagggg acactttcta
      901 taataaagtt cgattttatg gtaccaactt tcccgccaat ggtccagtta tgcagaagaa
      961 gacgctgaaa tgggagccct ccactgagaa aatgtatgtg cgtgatggag tgctgacggg
     1021 tgatattgag atggctttgt tgcttgaagg aggtgcccat taccgatgtg acttcagaac
     1081 tacttacaaa gctaaggaga agggtgtcaa gttaccaggc gcccactttg tggaccacgc
     1141 cattgagatt ttaagccatg acaaagatta caacaaggtt aagctgtatg agcatgctgt
     1201 tgctcattct ggattgcctg acaatgccag acgatccggc cggactcaga tctcgagctc
     1261 aagcttcgaa ttcatggaga ccccgtccca gcggcgcgcc acccgcagcg gggcgcaggc
     1321 cagctccact ccgctgtcgc ccacccgcat cacccggctg caggagaagg aggacctgca
     1381 ggagctcaat gatcgcttgg cggtctacat cgaccgtgtg cgctcgctgg aaacggagaa
     1441 cgcagggctg cgccttcgca tcaccgagtc tgaagaggtg gtcagccgcg aggtgtccgg
     1501 catcaaggcc gcctacgagg ccgagctcgg ggatgcccgc aagacccttg actcagtagc
     1561 caaggagcgc gcccgcctgc agctggagct gagcaaagtg cgtgaggagt ttaaggagct
     1621 gaaagcgcgc aataccaaga aggagggtga cctgatagct gctcaggctc ggctgaagga
     1681 cctggaggct ctgctgaact ccaaggaggc cgcactgagc actgctctca gtgagaagcg
     1741 cacgctggag ggcgagctgc atgatctgcg gggccaggtg gccaagcttg aggcagccct
     1801 aggtgaggcc aagaagcaac ttcaggatga gatgctgcgg cgggtggatg ctgagaacag
     1861 gctgcagacc atgaaggagg aactggactt ccagaagaac atctacagtg aggagctgcg
     1921 tgagaccaag cgccgtcatg agacccgact ggtggagatt gacaatggga agcagcgtga
     1981 gtttgagagc cggctggcgg atgcgctgca ggaactgcgg gcccagcatg aggaccaggt
     2041 ggagcagtat aagaaggagc tggagaagac ttattctgcc aagctggaca atgccaggca
     2101 gtctgctgag aggaacagca acctggtggg ggctgcccac gaggagctgc agcagtcgcg
     2161 catccgcatc gacagcctct ctgcccagct cagccagctc cagaagcagc tggcagccaa
     2221 ggaggcaaag cttcgagacc tggaggactc actggcccgt gagcgggaca ccagccggcg
     2281 gctgctggcg gaaaaggagc gggagatggc cgagatgcgg gcaaggatgc agcagcagct
     2341 ggacgagtac caggagcttc tggacatcaa gctggccctg gacatggaga tccacgccta
     2401 ccgcaagctc ttggagggcg aggaggagag gctacgcctg tcccccagcc ctacctcgca
     2461 gcgcagccgt ggccgtgctt cctctcactc atcccagaca cagggtgggg gcagcgtcac
     2521 caaaaagcgc aaactggagt ccactgagag ccgcagcagc ttctcacagc acgcacgcac
     2581 tagcgggcgc gtggccgtgg aggaggtgga cgaggagggc aagtttgtcc ggctgcgcaa
     2641 caagtccaat gaggaccagt ccatgggcaa ttggcagatc aagcgccaga atggagatga
     2701 tcccttgctg acttaccggt tcccaccaaa gttcaccctg aaggctgggc aggtggtgac
     2761 gatctgggct gcaggagctg gggccaccca cagcccccct accgacctgg tgtggaaggc
     2821 acagaacacc tggggctgcg ggaacagcct gcgtacggct ctcatcaact ccactgggga
     2881 agaagtggcc atgcgcaagc tggtgcgctc agtgactgtg gttgaggacg acgaggatga
     2941 ggatggagat gacctgctcc atcaccacca cggctcccac tgcagcagct cgggggaccc
     3001 cgctgagtac aacctgcgct cgcgcaccgt gctgtgcggg acctgcgggc agcctgccga
     3061 caaggcatct gccagcggct caggagccca ggtgggcgga cccatctcct ctggctcttc
     3121 tgcctccagt gtcacggtca ctcgcagcta ccgcagtgtg gggggcagtg ggggtggcag
     3181 cttcggggac aatctggtca cccgctccta cctcctgggc aactccagcc cccgaaccca
     3241 gagcccccag aactgcagca tcatgtaagt cgacggtacc gcgggcccgg gatccaccgg
     3301 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3361 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3421 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3481 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3541 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3601 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3661 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3721 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3781 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3841 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3901 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3961 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4141 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4201 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4321 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4381 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4441 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4501 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4561 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4621 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4681 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4741 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4801 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4861 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4921 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4981 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5041 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5101 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5161 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5221 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5281 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5341 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5401 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5461 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5521 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5581 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5641 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5701 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5761 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5821 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5881 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5941 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6001 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6061 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6121 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6181 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6241 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6301 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6361 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6421 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6481 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6541 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6601 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     6661 cattagttca tagcccatat atggagttcc
//
