LOCUS       sequence-190908-        6635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the nuclear envelope (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6635)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 6635)
  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"; journalName: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: mScarlet-I-LaminB
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1308
                     /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
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(746..764)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1014..1033
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1348..1389
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    1405..1429
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3227..3251)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3423..3544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3460..3479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3514..3533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3551..4006)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3688..3709)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3900..3919
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4033..4137
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4139..4496
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4139..4330
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4162..4182)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4347..4482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4531..5325
                     /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             4531..5325
                     /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(4585..4604)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5195..5214
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5513..5532)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5557..5604
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5933..6521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6422..6441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag
      661 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
      721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      781 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac
      841 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc
      901 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac
      961 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta
     1021 atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc
     1081 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg
     1141 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc
     1201 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa
     1261 cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
     1321 tctcgagctc aagcttcgaa ttgcggtggt aagcctatcc ctaaccctct cctcggtctc
     1381 gattctacga tcgatttcga tcaaacaagt ttgtacaaaa aagcaggctc cgcggccgcc
     1441 cccttcacca tggcgactgc gacccccgtg ccgccgcgga tgggcagccg cgctggcggc
     1501 cccaccacgc cgctgagccc cacgcgcctg tcgcggctcc aggagaagga ggagctgcgc
     1561 gagctcaatg accggctggc ggtgtacatc gacaaggtgc gcagcctgga gacggagaac
     1621 agcgcgctgc agctgcaggt gacggagcgc gaggaggtgc gcggccgtga gctcaccggc
     1681 ctcaaggcgc tctacgagac cgagctggcc gacgcgcgac gcgcgctcga cgacacggcc
     1741 cgcgagcgcg ccaagctgca gatcgagctg ggcaagtgca aggcggaaca cgaccagctg
     1801 ctcctcaact atgctaagaa ggaatctgat cttaatggcg cccagatcaa gcttcgagaa
     1861 tatgaagcag cactgaattc gaaagatgca gctcttgcta ctgcacttgg tgacaaaaaa
     1921 agtttagagg gagatttgga ggatctgaag gatcagattg cccagttgga agcctcctta
     1981 gctgcagcca aaaaacagtt agcagatgaa actttactta aagtagattt ggagaatcgt
     2041 tgtcagagcc ttactgagga cttggagttt cgcaaaagca tgtatgaaga ggagattaac
     2101 gagaccagaa ggaagcatga aacgcgcttg gtagaggtgg attctgggcg tcaaattgag
     2161 tatgagtaca agctggcgca agcccttcat gagatgagag agcaacatga tgcccaagtg
     2221 aggctgtata aggaggagct ggagcagact taccatgcca aacttgagaa tgccagactg
     2281 tcatcagaga tgaatacttc tactgtcaac agtgccaggg aagaactgat ggaaagccgc
     2341 atgagaattg agagcctttc atcccagctt tctaatctac agaaagagtc tagagcatgt
     2401 ttggaaagga ttcaagaatt agaggacttg cttgctaaag aaaaagacaa ctctcgtcgc
     2461 atgctgacag acaaagagag agagatggcg gaaataaggg atcaaatgca gcaacagctg
     2521 aatgactatg aacagcttct tgatgtaaag ttagccctgg acatggaaat cagtgcttac
     2581 aggaaactct tacaaggcga agaagagagg ttgaagctgt ctccaagccc ttcttcccgt
     2641 gtgacagtat cccgagcatc ctcaagtcgt agtgtacgta caactagagg aaagcggaag
     2701 agggttgatg tggaagaatc agaggcgagt agtagtgtta gcatctctca ttccgcctca
     2761 gccactggaa atgtttgcat cgaagaaatt gatgttgatg ggaaatttat ccgcttgaag
     2821 aacacttctg aacaggatca accaatggga ggctgggaga tgatcagaaa aattggagac
     2881 acatcagtca gttataaata tacctcaaga tatgtgctga aggcaggcca gactgttaca
     2941 atttgggctg caaacgctgg tgtcacagcc agccccccaa ctgacctcat ctggaagaac
     3001 cagaactcgt ggggcactgg cgaagatgtg aaggttatat tgaaaaattc tcagggagag
     3061 gaggttgctc aaagaagtac agtctttaaa acaaccatac ctgaagaaga ggaggaggag
     3121 gaagaagcag ctggagtggt tgttgaggaa gaacttttcc accagcaggg aaccccaaga
     3181 gcatccaata gaagctgtgc aattatgtaa aagggtgggc gcgccgaccc agctttcttg
     3241 tacaaagtgg ttcgatcgaa ccgcggtcta gactcgacgg taccgcgggc ccgggatcca
     3301 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3361 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3421 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3541 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3661 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3721 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3781 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3841 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3901 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3961 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4021 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4081 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4141 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4201 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4321 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4381 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4441 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4501 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4561 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4621 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4681 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4741 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4801 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4861 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4921 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4981 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5041 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5101 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5161 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5221 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5281 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5341 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5401 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5461 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5521 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5581 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5641 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5701 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5761 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6061 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6601 ctgattctgt ggataaccgt attaccgcca tgcat
//
