LOCUS       sequence-193715-        4092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Ferritin-Dronpa fusion construct in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4092)
  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 4092)
  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-Ferritin-Dronpa
FEATURES             Location/Qualifiers
     source          1..4092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..40
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     125..144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        125..143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             175..197
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             217..234
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             238..270
                     /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(274..282,283..297)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             868..1542
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     primer_bind     complement(1598..1616)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1612..1659
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1756..2211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1843..1862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2053..2074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2237..2341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2342..2410,2411..3202)
                     /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(2560..2579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3373..3961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3862..3881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
       61 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct cgatcccgcg
      121 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
      181 taactttaag aaggagatat acatatgcgg ggttctcatc atcatcatca tcatggtatg
      241 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
      301 ccgatggact cacaggtgcg ccagaatttc catcgtgatt gcgaggccgc aattaatcgc
      361 atggtcaaca tggaactgta tgcctcgtat acctatctca gcatggcatt ttattttgat
      421 cgcgatgaca tcgcgttaca caacgtagct aaattcttta aggaacaatc tcatgaagaa
      481 cgtgaacacg cggaaaaact tatgaaagac caaaataaac gtggtggccg tattgtattg
      541 caggatgttg aaaaaccgga acgtgacgag tggggtaata cgttggaggc gatgcaggcc
      601 gcgttacagt tagagaaaac agtgaatcag gccctgcttg atctgcataa agttggttcc
      661 gataaagtcg atccgcactt atgtgatttc ttggagaccg aatatttaga agaacaggtg
      721 aaatcgatta aacaattagg ggattacatt acgaacctga aacgtctggg actgccgcag
      781 aacggaatgg gcgaatatct gtttgataaa cacaccatgg gggaatcctc tggtaccgct
      841 ggctccgctg ctggttctgg cgaattcatg agtgtgatta aaccagacat gaagatcaag
      901 ctgcgtatgg aaggcgctgt aaatggacac ccgttcgcga ttgaaggagt tggccttggg
      961 aagcctttcg agggaaaaca gagtatggac cttaaagtca aagaaggcgg acctctgcct
     1021 ttcgcctatg acatcttgac aactgtgttc tgttacggca acagggtatt cgccaaatac
     1081 ccagaaaata tagtagacta tttcaagcag tcgtttcctg agggctactc ttgggaacga
     1141 agcatgaatt acgaagacgg gggcatttgt aacgcgacaa acgacataac cctggatggt
     1201 gactgttata tctatgaaat tcgatttgat ggtgtgaact ttcctgccaa tggtccagtt
     1261 atgcagaaga ggactgtgaa atgggagcca tccactgaga aattgtatgt gcgtgatgga
     1321 gtgctgaagg gtgatgttaa catggctctg tcgcttgaag gaggtggcca ttaccgatgt
     1381 gacttcaaaa ctacttataa agctaagaag gttgtccagt tgccagacta tcactttgtg
     1441 gaccaccaca ttgagattaa aagccacgac aaagattaca gtaatgttaa tctgcatgag
     1501 catgccgaag cgcattctga gctgccgagg caggccaagt aaaagcttga tccggctgct
     1561 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1621 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1681 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1741 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1801 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1861 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1921 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1981 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2041 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2101 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2161 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2221 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2281 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2341 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2401 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2461 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2521 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2581 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2641 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2701 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2761 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2821 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2881 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2941 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3001 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3061 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3121 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3181 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3241 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3301 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3361 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3421 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3481 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3541 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3601 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3661 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3721 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3781 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3841 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3901 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3961 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4021 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4081 tgataccgct cg
//
