LOCUS       sequence-193714-        3535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A fast switching Dronpa variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3535)
  AUTHORS   Duwe S, Vandenberg W, Dedecker P
  TITLE     Live-cell monochromatic dual-label sub-diffraction microscopy by 
            mt-pcSOFI.
  JOURNAL   Chem Commun (Camb). 2017 Jun 29;53(53):7242-7245. doi: 
            10.1039/c7cc02344h.
  PUBMED    28561832
REFERENCE   2  (bases 1 to 3535)
  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"; doi: "10"; journalName: 
            "Chem Commun (Camb)"; date: "2017-06-29- 29"; volume: "53"; issue: 
            "53"; pages: "7242-7245"
COMMENT     Sequence Label: pRSETb-ffDronpa-F
FEATURES             Location/Qualifiers
     source          1..3535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        73..91
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             165..182
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             186..218
                     /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(222..230,231..245)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             252..926
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVI
                     EGGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDVKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     primer_bind     complement(989..1007)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1003..1050
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1147..1602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1234..1253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1444..1465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1628..1732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1733..1801,1802..2593)
                     /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(1951..1970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2764..3352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3253..3272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3506..3523
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggatctcg
       61 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
      121 attttgttta actttaagaa ggagatatac atatgcgggg ttctcatcat catcatcatc
      181 atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg
      241 ataaggatcc gatgagtgtg attaaaccag acatgaagat caagctgcgt atggaaggcg
      301 ctgtaaatgg acacccgttc gcgattgaag gagttggcct tgggaagcct ttcgagggaa
      361 aacagagtat ggaccttaaa gtcatagagg gcggacctct gcctttcgcc tatgacatct
      421 tgacaactgc gttctgttac ggcaacaggg tattcgccaa atacccagaa aatatagtag
      481 actatttcaa gcagtcgttt cctgagggct actcttggga acgaagcatg aattacgaag
      541 acgggggcat ttgtaacgcg acaaacgaca taaccctgga tggtgactgt tatatctatg
      601 aaattcgatt tgatggtgtg aactttcctg ccaatggtcc agttatgcag aagaggactg
      661 tgaaatggga gccatccact gagaaattgt atgtgcgtga tggagtgctg aagggcgatg
      721 ttaacatggc tctgtcgctt gaaggaggtg gccattaccg atgtgacgtg aaaactactt
      781 ataaagctaa gaaggttgtc cagttgccag actatcactt tgtggaccac cacattgaga
      841 ttaaaagcca cgacaaagat tacagtaatg ttaatctgca tgagcatgcc gaagcgcatt
      901 ctgagctgcc gaggcaggcc aagtaagaat tcgaagcttg atccggctgc taacaaagcc
      961 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1021 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctggc
     1081 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1141 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1501 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1561 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     1621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagag
//
