LOCUS       sequence-145796-        3535 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses ffDronpa E. coli strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3535)
  AUTHORS   Moeyaert B, Nguyen Bich N, De Zitter E, Rocha S, Clays K, Mizuno H, 
            van Meervelt L, Hofkens J, Dedecker P
  TITLE     Green-to-red photoconvertible Dronpa mutant for multimodal 
            super-resolution fluorescence microscopy.
  JOURNAL   ACS Nano. 2014 Feb 25;8(2):1664-73. doi: 10.1021/nn4060144. Epub 
            2014 Jan 21.
  PUBMED    24410188
REFERENCE   2  (bases 1 to 3535)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/nn4060144";
            journalName: "ACS Nano"; date: "2014-02-25- 25"; volume: "8"; issue:
            "2"; pages: "1664-73"
COMMENT     Sequence Label: pRSETb-ffDronpa
FEATURES             Location/Qualifiers
     source          1..3535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /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(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             199..873
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     primer_bind     complement(936..954)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      950..997
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1094..1549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1181..1200)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1391..1412
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1575..1679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1680..1748,1749..2540)
                     /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(1898..1917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2711..3299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3200..3219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3453..3470
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccgat gagtgtgatt aaaccagaca tgaagatcaa gctgcgtatg
      241 gaaggcgctg taaatggaca cccgttcgcg attgaaggag ttggccttgg gaagcctttc
      301 gagggaaaac agagtatgga ccttaaagtc aaagaaggcg gacctctgcc tttcgcctat
      361 gacatcttga caactgcgtt ctgttacggc aacagggtat tcgccaaata cccagaaaat
      421 atagtagact atttcaagca gtcgtttcct gagggctact cttgggaacg aagcatgaat
      481 tacgaagacg ggggcatttg taacgcgaca aacgacataa ccctggatgg tgactgttat
      541 atctatgaaa ttcgatttga tggtgtgaac tttcctgcca atggtccagt tatgcagaag
      601 aggactgtga aatgggagcc atccactgag aaattgtatg tgcgtgatgg agtgctgaag
      661 ggtgatgtta acatggctct gtcgcttgaa ggaggtggcc attaccgatg tgacttcaaa
      721 actacttata aagctaagaa ggttgtccag ttgccagact atcactttgt ggaccaccac
      781 attgagatta aaagccacga caaagattac agtaatgtta atctgcatga gcatgccgaa
      841 gcgcattctg agctgccgag gcaggccaag taagaattcg aagcttgatc cggctgctaa
      901 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
      961 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1021 atctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1081 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1141 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1201 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1261 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1321 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1381 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1441 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1501 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     1561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1921 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2221 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2821 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3361 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3421 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3481 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcag
//
