LOCUS       sequence-203403-        3553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  photoconvertible fluorescent protein mEos4b-V69T in pRSetA plasmid, 
            photoconvertible by primed conversion mechanism, bacterial 
            overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3553)
  AUTHORS   Turkowyd B, Balinovic A, Virant D, Golz Carnero HG, Caldana F, 
            Endesfelder M, Bourgeois D, Endesfelder U
  TITLE     A general mechanism of photoconversion of green-to-red fluorescent 
            proteins based on blue and infrared light reduces phototoxicity in 
            live-cell single-molecule imaging.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Jun 2. doi: 10.1002/anie.201702870.
  PUBMED    28574633
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: "Angew Chem 
            Int Ed Engl. 2017 Jun 2. doi: 10.1002/anie.201702870."
COMMENT     Sequence Label: pRSETa-mEos4b-V69T
FEATURES             Location/Qualifiers
     source          1..3553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             69..91
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             111..128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             132..164
                     /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(168..176,177..191)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             207..890
                     /codon_start=1
                     /product="fixation-resistant truly monomeric green-to-red 
                     photoswitchable fluorescent protein (Paez-Segala et al., 
                     2015)"
                     /label=mEos4b
                     /note="derived from mEos2"
                     /translation="MVSAIKPDMRIKLRMEGNVNGHHFVIDGDGTGKPYEGKQTMDLEV
                     KEGGPLPFAFDILTTAFHYGNRVFTKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNA
                     RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL
                     LEGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     primer_bind     complement(953..971)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      967..1014
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1111..1566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1198..1217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1408..1429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1592..1696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1697..1765,1766..2557)
                     /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(1915..1934)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2728..3316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3217..3236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3470..3487
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc
      121 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg
      181 atgacgataa ggatcgatgg ggatccatgg tgagtgcgat taagccagac atgaggatca
      241 aactccgtat ggaaggcaac gtaaacgggc accactttgt gatcgacgga gatggtacag
      301 gcaagcctta tgagggaaaa cagaccatgg atcttgaagt caaagagggc ggacctctgc
      361 cttttgcctt tgatatcctg accactgcat tccattacgg caacagggta ttcacgaaat
      421 atccagacaa catacaagac tattttaagc agtcgtttcc taaggggtat tcgtgggaac
      481 gaagcttgac tttcgaagac gggggcattt gcaatgccag aaacgacata acaatggaag
      541 gggacacttt ctataataaa gttcgatttt atggtaccaa ctttcccgcc aatggtccag
      601 ttatgcagaa gaagacgctg aaatgggagc cctccactga gaaaatgtat gtgcgtgatg
      661 gagtgctgac gggtgatatt gagatggctt tgttgcttga aggaaatgcc cattaccgat
      721 gtgacttcag aactacttac aaagctaagg agaagggtgt caagttacca ggcgcccact
      781 ttgtggacca cgccattgag attttaagcc atgacaaaga ttacaacaag gttaagctgt
      841 atgagcatgc tgttgctcat tctggattgc ctgacaatgc cagacgataa gaattcgaag
      901 cttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
      961 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1021 ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1081 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     1141 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1201 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1261 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1321 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1381 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1441 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1501 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     1561 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1621 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1681 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1741 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1801 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1861 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1921 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1981 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2041 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2101 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2161 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2221 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2281 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2341 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2401 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2461 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2521 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2581 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2641 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3421 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3481 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3541 tcattaatgc agg
//
