LOCUS       sequence-376497-        4828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for testing oligimerization of mScarlet-I3-NCwt in 
            mammalian cells by OSER assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4828)
  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 4828)
  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-mScarlet-I3-NCwt_N1
FEATURES             Location/Qualifiers
     source          1..4828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             66..860
                     /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             66..860
                     /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(120..139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     730..749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1048..1067)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1092..1139
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1468..2056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1957..1976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2231..2534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2535..2738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2689..2709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2960..2969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2966..3661
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGG
                     TVSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARH
                     STGGMDELYK"
     primer_bind     complement(3099..3117)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3786..3907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3823..3842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3877..3896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3914..4369)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4051..4072)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4263..4282
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4396..4500
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4502..4828,1..31)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4502..4693
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4525..4545)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4710..4828,1..17)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
       61 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      121 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      181 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      241 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      301 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      361 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      421 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
      481 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
      541 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
      601 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
      661 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
      721 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
      781 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
      841 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
      901 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
      961 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1021 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     1081 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     1141 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     1201 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     1261 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     1321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2161 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2701 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     2761 gctagcgcta ccggactcag atctggtacc ccttgcagct tccagttgtc aggaagagaa
     2821 ggctcccatg gatcctgtgg tggtgctggg gctctgtctc tcctgtttgc ttctcctttc
     2881 actctggaaa cagagctatg ggggagggaa gcttcgaatt ctgcagtcga cggtaccgcg
     2941 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggcagtga tcaaggagtt
     3001 catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg
     3061 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     3121 tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc
     3181 cttcatcaag caccccgccg acatccccga ctactggaag cagtccttcc ccgagggctt
     3241 caagtgggag cgcgtgatga tcttcgagga cggcggcacc gtgtccgtga cccaggacac
     3301 ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcggca acttccctcc
     3361 tgacggcccc gtaatgcaga agaggacaat gggctgggaa gcatccaccg agcggttgta
     3421 ccccgaggac gtcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg
     3481 ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg
     3541 cgccttcaac atcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt
     3601 ggaacagtac gaacgctccg tggcccgcca ctccaccggc ggcatggacg agctgtacaa
     3661 gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3721 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3781 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3841 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3901 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3961 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4021 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4081 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4141 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4201 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4261 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4321 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4381 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4441 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4501 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4561 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4621 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4681 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4741 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4801 cctctgagct attccagaag tagtgagg
//
