LOCUS       sequence-376494-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for labelling nuclear envelopes in mammaiian cells using 
            mScarlet3-LaminB fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  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 6634)
  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: pmScarlet3-LaminB_C1
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      720..729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             726..1412
                     /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     1124..1143
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1461..1502
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    1518..1542
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3340..3364)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3536..3657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3573..3592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3627..3646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3664..4119)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3801..3822)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4013..4032
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4146..4250
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4252..4609
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4252..4443
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4275..4295)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4460..4595
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4644..5438
                     /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             4644..5438
                     /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(4698..4717)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5308..5327
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5626..5645)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5670..5717
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6046..6634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6535..6554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtg
      721 ccaccatgga tagcaccgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
      781 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
      841 agggcaccca gaccgccaag ctgagggtga ccaagggtgg ccccctgccc ttctcctggg
      901 acatcctgtc ccctcagttc atgtacggct ccagggcctt cacgaagcac cccgccgaca
      961 tccccgacta ctggaagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1021 tcgaggacgg cggcgccgtg tccgtggccc aggacacctc cctggaggac ggcaccctga
     1081 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     1141 agacaatggg ctgggaagca tccaccgagc ggttgtaccc cgaggacgtc gtgctgaagg
     1201 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     1261 ccacctacag ggccaagaag cccgtgcaga tgcccggcgc cttcaacatc gaccgcaagt
     1321 tggacatcac atcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgtgg
     1381 cccgccactc caccggcggc tccggtggct ccttgtacaa gtccggactc agatctcgag
     1441 ctcaagcttc gaattgcggt ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     1501 cgatcgattt cgatcaaaca agtttgtaca aaaaagcagg ctccgcggcc gcccccttca
     1561 ccatggcgac tgcgaccccc gtgccgccgc ggatgggcag ccgcgctggc ggccccacca
     1621 cgccgctgag ccccacgcgc ctgtcgcggc tccaggagaa ggaggagctg cgcgagctca
     1681 atgaccggct ggcggtgtac atcgacaagg tgcgcagcct ggagacggag aacagcgcgc
     1741 tgcagctgca ggtgacggag cgcgaggagg tgcgcggccg tgagctcacc ggcctcaagg
     1801 cgctctacga gaccgagctg gccgacgcgc gacgcgcgct cgacgacacg gcccgcgagc
     1861 gcgccaagct gcagatcgag ctgggcaagt gcaaggcgga acacgaccag ctgctcctca
     1921 actatgctaa gaaggaatct gatcttaatg gcgcccagat caagcttcga gaatatgaag
     1981 cagcactgaa ttcgaaagat gcagctcttg ctactgcact tggtgacaaa aaaagtttag
     2041 agggagattt ggaggatctg aaggatcaga ttgcccagtt ggaagcctcc ttagctgcag
     2101 ccaaaaaaca gttagcagat gaaactttac ttaaagtaga tttggagaat cgttgtcaga
     2161 gccttactga ggacttggag tttcgcaaaa gcatgtatga agaggagatt aacgagacca
     2221 gaaggaagca tgaaacgcgc ttggtagagg tggattctgg gcgtcaaatt gagtatgagt
     2281 acaagctggc gcaagccctt catgagatga gagagcaaca tgatgcccaa gtgaggctgt
     2341 ataaggagga gctggagcag acttaccatg ccaaacttga gaatgccaga ctgtcatcag
     2401 agatgaatac ttctactgtc aacagtgcca gggaagaact gatggaaagc cgcatgagaa
     2461 ttgagagcct ttcatcccag ctttctaatc tacagaaaga gtctagagca tgtttggaaa
     2521 ggattcaaga attagaggac ttgcttgcta aagaaaaaga caactctcgt cgcatgctga
     2581 cagacaaaga gagagagatg gcggaaataa gggatcaaat gcagcaacag ctgaatgact
     2641 atgaacagct tcttgatgta aagttagccc tggacatgga aatcagtgct tacaggaaac
     2701 tcttacaagg cgaagaagag aggttgaagc tgtctccaag cccttcttcc cgtgtgacag
     2761 tatcccgagc atcctcaagt cgtagtgtac gtacaactag aggaaagcgg aagagggttg
     2821 atgtggaaga atcagaggcg agtagtagtg ttagcatctc tcattccgcc tcagccactg
     2881 gaaatgtttg catcgaagaa attgatgttg atgggaaatt tatccgcttg aagaacactt
     2941 ctgaacagga tcaaccaatg ggaggctggg agatgatcag aaaaattgga gacacatcag
     3001 tcagttataa atatacctca agatatgtgc tgaaggcagg ccagactgtt acaatttggg
     3061 ctgcaaacgc tggtgtcaca gccagccccc caactgacct catctggaag aaccagaact
     3121 cgtggggcac tggcgaagat gtgaaggtta tattgaaaaa ttctcaggga gaggaggttg
     3181 ctcaaagaag tacagtcttt aaaacaacca tacctgaaga agaggaggag gaggaagaag
     3241 cagctggagt ggttgttgag gaagaacttt tccaccagca gggaacccca agagcatcca
     3301 atagaagctg tgcaattatg taaaagggtg ggcgcgccga cccagctttc ttgtacaaag
     3361 tggttcgatc gaaccgcggt ctagactcga cggtaccgcg ggcccgggat ccaccggatc
     3421 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3481 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3541 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3601 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3661 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3721 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3781 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3841 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3901 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3961 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4021 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4081 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4201 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4261 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4321 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4381 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4441 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4501 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4561 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4621 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4681 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4741 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4801 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4861 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4921 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4981 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5041 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5101 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5161 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5221 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5281 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5341 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5401 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5461 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5521 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5581 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5641 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5701 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5761 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5821 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5881 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5941 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6001 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6061 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6121 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6181 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6421 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6601 gtgatgctcg tcaggggggc ggagcctatg gaaa
//
