LOCUS       sequence-194796-        6940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the photoconvertible protein mEos2 as an 
            Ensconsin fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6940)
  AUTHORS   Mohr MA, Kobitski AY, Sabater LR, Nienhaus K, Obara CJ, 
            Lippincott-Schwartz J, Nienhaus GU, Pantazis P
  TITLE     Rational Engineering of Photoconvertible Fluorescent Proteins for 
            Dual-Color Fluorescence Nanoscopy Enabled by a Triplet-State 
            Mechanism of Primed Conversion.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Jun 29. doi: 10.1002/anie.201706121.
  PUBMED    28661566
REFERENCE   2  (bases 1 to 6940)
  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: "Angew Chem 
            Int Ed Engl. 2017 Jun 29. doi: 10.1002/anie.201706121."
COMMENT     Sequence Label: Ensconsin-18-mEos2
FEATURES             Location/Qualifiers
     source          1..6940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2866..3546
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    3668..3789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3705..3724)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3759..3778
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3796..4251)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3933..3954)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4145..4164
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4278..4382
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4384..4741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4384..4575
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4407..4427)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4592..4727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4776..5570
                     /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             4776..5570
                     /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(4830..4849)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5440..5459
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5758..5777)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5802..5849
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6178..6766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6667..6686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgaggc caccatggcg gagctaggag ctggcggcga cggccacagg
      601 ggcggcgacg gcgcagtgcg aagcgaaaca gcacccgaca gctacaaagt gcaagataag
      661 aaaaatgcct ccagccgccc tgcctctgca atttcaggac aaaataacaa ccactcagga
      721 aataaaccag accctccgcc tgtgttacgt gttgatgacc ggcagcggct ggcccgggag
      781 cgacgtgagg aacgggagaa acagctagct gcaagagaaa tagtgtggtt agaaagagaa
      841 gagcgagcca ggcagcacta cgagaagcac ctggaagagc ggaagaagag gttggaggag
      901 cagaggcaga aggaggagcg gaggagggct gctgtggagg agaagcggag gcagagactt
      961 gaggaggaca aagaacgcca cgaagctgtt gtacggcgca caatggaaag gagccagaag
     1021 ccaaaacaga agcataaccg ttggtcgtgg ggaggctctc tccatgggag ccctagcatc
     1081 cacagtgcag atccagacag gcggtcagtt tccaccatga atctttcgaa atatgttgat
     1141 cccgtcatta gcaagcggct ctcctcttca tctgcaactt tactaaattc tccagataga
     1201 gctcgccgcc tgcagctcag cccatgggag agcagcgttg ttaacagact cctgacgccc
     1261 acacattcgt tcctggccag aagtaaaagc acagctgcct tgtctggaga agcagcatct
     1321 tgcagcccca tcatcatgcc ctacaaagct gcacactcta gaaattcgat ggatcgacca
     1381 aaactctttg taacaccacc tgagggctct tctcgcagga ggatcattca tggcacagcg
     1441 agctataaaa aagaaagaga gagagaaaat gtactcttcc tcacatctgg cacccgaagg
     1501 gctgtatctc catctaatcc caaagcaaga caaccagctc gctcccgact ttggcttccg
     1561 tccaagtctc ttcctcattt gcctggcaca cccagaccga catcctcctt gccacccggc
     1621 tcagtcaaag ctgctcctgc tcaggtccgg cccccatccc ccggcaacat ccgccctgtc
     1681 aagagggaag tcaaagtgga gcctgagaag aaagatcctg agaaggaacc tcagaaagtt
     1741 gccaatgagc cctcactaaa gggcagagca cctttagtga aggtagaaga agccacagtt
     1801 gaagagcgga cacctgctga accagaagtt ggccctgctg ctccagccat ggccccagct
     1861 ccagcctcgg ccccagctcc agcctcggcc ccagctccag ccccggtccc caccccagcc
     1921 atggtctcag ccccgtcatc cactgtgaat gccagtgctt ctgttaagac ttctgcaggc
     1981 accaccgacc cagaggaggc cacaaggctt ctagctgaga agaggcggct ggcccgagag
     2041 cagagagaaa aggaagaaag ggagaggagg gagcaggaag agcttgaaag acaaaagaga
     2101 gaggaattgg ctcaacgtgt ggctgaagag aggacgactc gccgtgagga ggagtcgcgc
     2161 aggctggaag ccgagcaggc ccgggagaag gaggagcagc tgcagcggca ggcggaggag
     2221 cgggcgctgc gcgagtggga ggaggcagag cgcgcccaga ggcagaaaga agaagaagct
     2281 cgcgttcgtg aagaagcaga gagggtccgg caggaacgag agaagcattt ccagagagaa
     2341 gagcaagagc gcctggagag aaagaagcga cttgaggaga ttatgaaaag aaccaggaga
     2401 acagaagcta cagataagaa aaccagtgat cagagaaacg gtgatatagc caagggagct
     2461 ctcactggag gaacagaggt gtctgcactt ccatgtacaa caaacgctcc gggaaatgga
     2521 aagccagttg gtagcccaca tgtggttacc tcacaccagt caaaagtgac agtggagagc
     2581 actcccgatt tggaaaaaca accaaatgaa aatggtgtat ctgttcagaa tgaaaatttt
     2641 gaagaaatta taaacttacc cattggatct aaaccatcca gattagatgt caccaacagt
     2701 gagagcccag aaattccttt gaatccaatt ttggcctttg atgatgaagg gacacttggg
     2761 cccctgcctc aggtagatgg tgttcagaca cagcagactg cagaagttat aggcagcgca
     2821 ggcggaagcg gggtaccgcg ggcccgggat ccaccggtcg ccaccatgag tgcgattaag
     2881 ccagacatga agatcaaact ccgtatggaa ggcaacgtaa acgggcacca ctttgtgatc
     2941 gacggagatg gtacaggcaa gccttttgag ggaaaacaga gtatggatct tgaagtcaaa
     3001 gagggcggac ctctgccttt tgcctttgat atcctgacca ctgcattcca ttacggcaac
     3061 agggtattcg ccaaatatcc agacaacata caagactatt ttaagcagtc gtttcctaag
     3121 gggtattcgt gggaacgaag cttgactttc gaagacgggg gcatttgcat tgccagaaac
     3181 gacataacaa tggaagggga cactttctat aataaagttc gattttatgg taccaacttt
     3241 cccgccaatg gtccagttat gcagaagaag acgctgaaat gggagccctc cactgagaaa
     3301 atgtatgtgc gtgatggagt gctgacgggt gatattcata tggctttgtt gcttgaagga
     3361 aatgcccatt accgatgtga cttcagaact acttacaaag ctaaggagaa gggtgtcaag
     3421 ttaccaggct accactttgt ggaccactgc attgagattt taagccatga caaagattac
     3481 aacaaggtta agctgtatga gcatgctgtt gctcattctg gattgcctga caatgccaga
     3541 cgataagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3601 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3661 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3721 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3781 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3841 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3901 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3961 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4021 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4081 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4141 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4201 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4261 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4321 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4381 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4441 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4501 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4561 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4621 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4681 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4741 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4801 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4861 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4921 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4981 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5041 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5101 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5161 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5221 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5281 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5341 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5401 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5461 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5521 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5581 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5641 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5701 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5761 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5821 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5881 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5941 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     6001 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     6061 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6121 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6181 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6241 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6301 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6361 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6421 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6481 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6541 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6601 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6661 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6721 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6781 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6841 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     6901 ttacggggtc attagttcat agcccatata tggagttccg
//
