LOCUS       sequence-324282-        6914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of F-actin (can be used for colocalization 
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6914)
  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 6914)
  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: 3xmScarletI_UtrCH
FEATURES             Location/Qualifiers
     source          1..6914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1245
                     /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
                     STGGMDELY"
     CDS             1264..1956
                     /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
                     STGGMDELY"
     CDS             1975..2670
                     /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"
     misc_feature    3466..3518
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3642..3763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3679..3698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3733..3752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3770..4225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4119..4138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4252..4356
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4358..4715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4358..4549
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4381..4401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4566..4701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4750..5544
                     /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             4750..5544
                     /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(4804..4823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5414..5433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5732..5751)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5776..5823
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6152..6740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6641..6660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag
      601 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
      661 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      721 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac
      781 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc
      841 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac
      901 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta
      961 atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc
     1021 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg
     1081 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc
     1141 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa
     1201 cgctccgagg gccgccactc caccggcggc atggacgagc tgtaccaggg tggcagcggt
     1261 ggcatggtga gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg
     1321 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1381 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg
     1441 gacatcctgt cccctcagtt catgtacggc tccagggcct tcatcaagca ccccgccgac
     1501 atccccgact actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1561 ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg
     1621 atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag
     1681 aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag
     1741 ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc ggacttcaag
     1801 accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag
     1861 ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag
     1921 ggccgccact ccaccggcgg catggacgag ctgtaccagg gtggcagcgg tggcatggtg
     1981 agcaagggcg aggcagtgat caaggagttc atgcggttca aggtgcacat ggagggctcc
     2041 atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2101 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttctcctg ggacatcctg
     2161 tcccctcagt tcatgtacgg ctccagggcc ttcatcaagc accccgccga catccccgac
     2221 tactataagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2281 ggcggcgccg tgaccgtgac ccaggacacc tccctggagg acggcaccct gatctacaag
     2341 gtgaagctcc gcggcaccaa cttccctcct gacggccccg taatgcagaa gaagacaatg
     2401 ggctgggaag cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt
     2461 aagatggccc tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa gaccacctac
     2521 aaggccaaga agcccgtgca gatgcccggc gcctacaacg tcgaccgcaa gttggacatc
     2581 acctcccaca acgaggacta caccgtggtg gaacagtacg aacgctccga gggccgccac
     2641 tccaccggcg gcatggacga gctgtacaag tccggaacca tggccaagta tggagaacat
     2701 gaagccagtc ctgacaatgg gcagaacgaa ttcagtgata tcattaagtc cagatctgat
     2761 gaacacaatg acgtacagaa gaaaaccttt accaaatgga taaatgctcg attttcaaag
     2821 agtgggaaac cacccatcaa tgatatgttc acagacctca aagatggaag gaagctattg
     2881 gatcttctag aaggcctcac aggaacatca ctgccaaagg aacgtggttc cacaagggta
     2941 catgccttaa ataacgtcaa cagagtgctg caggttttac atcagaacaa tgtggaatta
     3001 gtgaatatag ggggaaccga cattgtggat ggaaatcaca aactgacttt ggggttactt
     3061 tggagcatca ttttgcactg gcaggtgaaa gatgtcatga aggatgtcat gtcggacctg
     3121 cagcagacga acagtgagaa gatcctgctc agctgggtgc gtcagaccac caggccctac
     3181 agccaagtca acgtcctcaa cttcaccacc agctggacag atggactcgc ctttaatgct
     3241 gtcctccacc gacataaacc tgatctcttc agctgggata aagttgtcaa aatgtcacca
     3301 attgagagac ttgaacatgc cttcagcaag gctcaaactt atttgggaat tgaaaagctg
     3361 ttagatcctg aagatgttgc cgttcggctt cctgacaaga aatccataat tatgtattta
     3421 acatctttgt ttgaggtgct acctcagcaa gtcaccatag actaactcga gctcaagctt
     3481 cgaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca
     3541 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3601 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3661 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3721 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3781 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3841 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3901 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3961 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4021 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4081 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4141 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4201 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4261 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4321 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4381 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4441 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4561 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4621 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4681 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4741 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4801 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4861 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4921 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4981 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5041 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5101 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5161 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5221 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5281 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5341 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5401 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5461 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5521 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5581 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5641 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5701 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5761 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5821 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5881 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5941 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     6001 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6061 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6121 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6181 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6241 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6301 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6361 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6421 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6481 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6541 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6601 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6661 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6721 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6781 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6841 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     6901 tatatggagt tccg
//
