LOCUS       sequence-376496-        4831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for testing oligimerization of mScarlet3 in mammalian 
            cells by OSER assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4831)
  AUTHORS   Gadella TWJ Jr, van Weeren L, Stouthamer J, Hink MA, Wolters AHG, 
            Giepmans BNG, Aumonier S, Dupuy J, Royant A
  TITLE     mScarlet3: a brilliant and fast-maturing red fluorescent protein.
  JOURNAL   Nat Methods. 2023 Apr;20(4):541-545. doi: 
            10.1038/s41592-023-01809-y. Epub 2023 Mar 27.
  PUBMED    36973546
REFERENCE   2  (bases 1 to 4831)
  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: 
            "10.1038/s41592-023-01809-y"; journalName: "Nat Methods"; date: 
            "2023-04"; volume: "20"; issue: "4"; pages: "541-545"
COMMENT     Sequence Label: pCyterm-mScarlet3_N1
FEATURES             Location/Qualifiers
     source          1..4831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             108..902
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             108..902
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(162..181)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     772..791
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1090..1109)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1134..1181
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1510..2098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1999..2018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2273..2576
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2577..2780
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2731..2751
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3002..3011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3008..3694
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     3406..3425
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3831..3952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3868..3887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3922..3941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3959..4414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4096..4117)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4308..4327
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4441..4545
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4547..4831,1..73)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4547..4738
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4570..4590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4755..4831,1..59)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
       61 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
      121 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
      181 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
      241 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
      301 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
      361 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
      421 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
      481 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
      541 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
      601 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
      661 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
      721 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
      781 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
      841 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
      901 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
      961 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     1021 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     1081 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     1141 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     1201 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     1261 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     1321 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     1381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1621 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1861 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1921 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1981 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2041 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2101 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2161 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     2221 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     2281 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2341 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2401 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     2461 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2521 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     2581 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     2641 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     2701 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     2761 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
     2821 agatctggta ccccttgcag cttccagttg tcaggaagag aaggctccca tggatcctgt
     2881 ggtggtgctg gggctctgtc tctcctgttt gcttctcctt tcactctgga aacagagcta
     2941 tgggggaggg aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt
     3001 cgccaccatg gatagcaccg aggcagtgat caaggagttc atgcggttca aggtgcacat
     3061 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     3121 cgagggcacc cagaccgcca agctgagggt gaccaagggt ggccccctgc ccttctcctg
     3181 ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcacgaagc accccgccga
     3241 catccccgac tactggaagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     3301 cttcgaggac ggcggcgccg tgtccgtggc ccaggacacc tccctggagg acggcaccct
     3361 gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg taatgcagaa
     3421 gaagacaatg ggctgggaag catccaccga gcggttgtac cccgaggacg tcgtgctgaa
     3481 gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa
     3541 gaccacctac agggccaaga agcccgtgca gatgcccggc gccttcaaca tcgaccgcaa
     3601 gttggacatc acatcccaca acgaggacta caccgtggtg gaacagtacg aacgctccgt
     3661 ggcccgccac tccaccggcg gctccggtgg ctcctaactg tacaagtaag cggccgcgac
     3721 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3781 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3841 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3901 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3961 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4021 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4081 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4141 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4201 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4261 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4321 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4381 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4441 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4501 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4561 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4621 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4741 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4801 gctgactaat tttttttatt tatgcagagg c
//
