LOCUS       sequence-495596-        3559 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Expression of mScarlet3-S2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3559)
  AUTHORS   Ding Y, He W, Wang K, Xue F, Zheng K, Zhao S, Qi D, Li T, Xia P, Pan
            L, Chen L, Chen Z, Yuan L, Xu P
  TITLE     A highly photostable monomeric red fluorescent protein for 
            dual-color 3D STED and time-lapse 3D SIM imaging.
  JOURNAL   Nat Methods. 2025 Nov 26. doi: 10.1038/s41592-025-02962-2.
  PUBMED    41299071
REFERENCE   2  (bases 1 to 3559)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2025 Nov 26. doi: 10.1038/s41592-025-02962-2."
COMMENT     Sequence Label: pRSETA-mScarlet3-S2
FEATURES             Location/Qualifiers
     source          1..3559
                     /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             208..894
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFQYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     606..625
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(960..978)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      974..1021
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1118..1573
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1205..1224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1415..1436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1599..1703
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1704..1772,1773..2564)
                     /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(1922..1941)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2735..3323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3224..3243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3477..3494
                     /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 aggatcgatg gggatccatg gatagcaccg aggcagtgat caaggagttc
      241 atgcggttca aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc
      301 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgagggt gaccaagggt
      361 ggccccctgc ccttctcctg ggacatcctg tcccctcagt tccagtacgg ctccagggcc
      421 ttcacgaagc accccgccga catccccgac tactggaagc agtccttccc cgagggcttc
      481 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg tgtccgtggc ccaggacacc
      541 tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct
      601 gacggccccg taatgcagaa gaagacaatg ggctgggaag catccaccga gcggttgtac
      661 cccgaggacg tcgtgctgaa gggcgacatt aagcacgccc tgcgcctgaa ggacggcggc
      721 cgctacctgg cggacttcaa gaccacctac agggccaaga agcccgtgca gatgcccggc
      781 gccttcaaca tcgaccgcaa gttggacatc acatcccaca acgaggacta caccgtggtg
      841 gaacagtacg aacgctccgt ggcccgccac tccaccggcg gctccggtgg ctcctaagaa
      901 ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
      961 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1021 gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc
     1081 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     1141 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1201 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1261 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1321 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1381 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1441 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1501 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1561 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1621 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1681 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1741 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1801 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1861 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1921 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1981 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2041 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2101 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2161 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2221 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2281 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2341 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2401 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2461 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2521 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2581 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2641 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2881 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3481 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     3541 ggccgattca ttaatgcag
//
