LOCUS       sequence-376488-        4790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for labelling endoplasmic reticulum in mammalian cells 
            using ER targeted mScarlet3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4790)
  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 4790)
  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: pER-mScarlet3_N1
FEATURES             Location/Qualifiers
     source          1..4790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             101..895
                     /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             101..895
                     /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(155..174)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     765..784
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1083..1102)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1127..1174
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1503..2091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1992..2011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2266..2569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2570..2773
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2724..2744
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     2846..2899
                     /codon_start=1
                     /product="signal sequence of chicken lysozyme"
                     /label=lysozyme signal sequence
                     /translation="MRSLLILVLCFLPLAALG"
     regulatory      2948..2957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2954..3640
                     /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     3352..3371
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3783..3904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3820..3839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3874..3893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3911..4366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4048..4069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4260..4279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4393..4497
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4499..4790,1..66)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4499..4690
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4522..4542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4707..4790,1..52)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
       61 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      121 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      181 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      241 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
      301 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      361 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      421 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      481 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      541 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      601 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
      661 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
      721 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
      781 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
      841 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
      901 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
      961 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1021 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     1081 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     1141 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     1201 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     1261 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     1321 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     1381 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1441 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1621 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1681 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1741 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1801 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1861 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1921 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1981 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2041 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2101 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2161 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     2221 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2281 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2341 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2401 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2461 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2521 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2581 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2641 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2701 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2761 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
     2821 gagctcaagc ttcgaattcg ccaccatgag gtctttgcta atcttggtgc tttgcttcct
     2881 gcccctggct gctctgggga aagtctttgg acgatgtgag ctggcagcgg ctatgaagcc
     2941 accggtcgcc accatggata gcaccgaggc agtgatcaag gagttcatgc ggttcaaggt
     3001 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3061 cccctacgag ggcacccaga ccgccaagct gagggtgacc aagggtggcc ccctgccctt
     3121 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca cgaagcaccc
     3181 cgccgacatc cccgactact ggaagcagtc cttccccgag ggcttcaagt gggagcgcgt
     3241 gatgaacttc gaggacggcg gcgccgtgtc cgtggcccag gacacctccc tggaggacgg
     3301 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
     3361 gcagaagaag acaatgggct gggaagcatc caccgagcgg ttgtaccccg aggacgtcgt
     3421 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga
     3481 cttcaagacc acctacaggg ccaagaagcc cgtgcagatg cccggcgcct tcaacatcga
     3541 ccgcaagttg gacatcacat cccacaacga ggactacacc gtggtggaac agtacgaacg
     3601 ctccgtggcc cgccactcca ccggcggctc cggtggctcc ttgtacaagg atgaattgta
     3661 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3721 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3781 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3841 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3901 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3961 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4021 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4081 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4141 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4201 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4261 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4321 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4501 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4621 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4681 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4741 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
//
