LOCUS       sequence-194794-        4689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the primed conversion capable protein 
            pr-mEos2 (mEos2-A69T) as a Lifeact fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4689)
  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 4689)
  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: pCMV-Lifect-7-pr-mEos2 (mEos2-A69T)
FEATURES             Location/Qualifiers
     source          1..4689
                     /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      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             543..593
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             615..1295
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFTKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    1417..1538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1454..1473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1508..1527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1545..2000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1682..1703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1894..1913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2027..2131
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2133..2490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2133..2324
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2156..2176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2341..2476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2525..3319
                     /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             2525..3319
                     /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(2579..2598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3189..3208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3507..3526)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3551..3598
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3927..4515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4416..4435
                     /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 gctagcgcca
      541 ccatgggcgt ggccgacttg atcaagaagt tcgagtccat ctccaaggag gagggggatc
      601 caccggtcgc caccatgagt gcgattaagc cagacatgaa gatcaaactc cgtatggaag
      661 gcaacgtaaa cgggcaccac tttgtgatcg acggagatgg tacaggcaag ccttttgagg
      721 gaaaacagag tatggatctt gaagtcaaag agggcggacc tctgcctttt gcctttgata
      781 tcctgaccac tgcattccat tacggcaaca gggtattcac caaatatcca gacaacatac
      841 aagactattt taagcagtcg tttcctaagg ggtattcgtg ggaacgaagc ttgactttcg
      901 aagacggggg catttgcatt gccagaaacg acataacaat ggaaggggac actttctata
      961 ataaagttcg attttatggt accaactttc ccgccaatgg tccagttatg cagaagaaga
     1021 cgctgaaatg ggagccctcc actgagaaaa tgtatgtgcg tgatggagtg ctgacgggtg
     1081 atattcatat ggctttgttg cttgaaggaa atgcccatta ccgatgtgac ttcagaacta
     1141 cttacaaagc taaggagaag ggtgtcaagt taccaggcta ccactttgtg gaccactgca
     1201 ttgagatttt aagccatgac aaagattaca acaaggttaa gctgtatgag catgctgttg
     1261 ctcattctgg attgcctgac aatgccagac gataagcggc cgcgactcta gatcataatc
     1321 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1381 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1441 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1501 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     1561 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     1621 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     1681 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     1741 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     1801 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     1861 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     1921 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     1981 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2041 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2101 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2161 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2221 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2281 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2341 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2401 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2461 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     2521 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2581 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2641 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2701 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2761 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2821 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     2881 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     2941 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3001 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3061 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3121 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3181 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3241 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3301 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3361 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3421 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3481 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3541 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     3601 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     3661 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     3721 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     3781 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3841 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3901 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3961 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4021 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4081 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4141 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4201 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4261 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4321 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4381 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4441 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4501 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4561 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4621 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     4681 ggagttccg
//
