LOCUS       sequence-377461-        4772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for labelling cell nuclei in mammalian cells using nuclear
            targeted mScarlet3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4772)
  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 4772)
  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: p3xnls-mScarlet3_N1
FEATURES             Location/Qualifiers
     source          1..4772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    140..187
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      516..1104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1005..1024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1279..1582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1583..1786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1737..1757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1870..1890
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1894..1914
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1918..1938
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1951..1960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1957..2643
                     /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     2355..2374
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2778..2899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2815..2834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2869..2888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2906..3361)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3043..3064)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3255..3274
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3388..3492
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3494..3851
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3494..3685
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3517..3537)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3702..3837
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3886..4680
                     /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             3886..4680
                     /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(3940..3959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4550..4569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
       61 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
      121 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
      181 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
      241 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
      301 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
      361 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
      421 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
      481 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      541 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      601 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      661 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      721 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      781 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      841 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      901 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      961 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1021 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1081 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1141 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1201 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     1261 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1561 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     1621 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1681 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1741 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     1801 tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat tcccaccatg
     1861 ggatcagatc caaaaaagaa gagaaaggta gacccgaaga aaaagcgtaa ggtcgatccc
     1921 aaaaagaaac ggaaagtggg atcaccggtc gccaccatgg atagcaccga ggcagtgatc
     1981 aaggagttca tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag
     2041 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgagggtg
     2101 accaagggtg gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc
     2161 tccagggcct tcacgaagca ccccgccgac atccccgact actggaagca gtccttcccc
     2221 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt gtccgtggcc
     2281 caggacacct ccctggagga cggcaccctg atctacaagg tgaagctccg cggcaccaac
     2341 ttccctcctg acggccccgt aatgcagaag aagacaatgg gctgggaagc atccaccgag
     2401 cggttgtacc ccgaggacgt cgtgctgaag ggcgacatta agatggccct gcgcctgaag
     2461 gacggcggcc gctacctggc ggacttcaag accacctaca gggccaagaa gcccgtgcag
     2521 atgcccggcg ccttcaacat cgaccgcaag ttggacatca catcccacaa cgaggactac
     2581 accgtggtgg aacagtacga acgctccgtg gcccgccact ccaccggcgg ctccggtggc
     2641 tccttgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2701 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2761 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2821 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2881 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2941 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3001 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3061 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3121 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3181 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3241 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3301 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3361 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3481 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3601 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3661 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3721 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3781 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3841 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3901 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3961 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4021 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4081 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4141 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4201 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4261 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4321 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4381 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4441 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4501 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4561 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4621 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4681 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4741 tcgattccac cgccgccttc tatgaaaggt tg
//
