LOCUS       sequence-224738-        6215 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 6215)
  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 6215)
  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: ITPKA_3xmScarletI
FEATURES             Location/Qualifiers
     source          1..6215
                     /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      754..763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             760..1452
                     /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             1471..2163
                     /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             2182..2877
                     /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"
     polyA_signal    3003..3124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3040..3059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3094..3113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3131..3586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3268..3289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3480..3499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3613..3717
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3719..4076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3719..3910
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3742..3762)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3927..4062
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4111..4905
                     /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             4111..4905
                     /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(4165..4184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4775..4794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5093..5112)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5137..5184
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5513..6101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6002..6021
                     /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 ccggactcag atctcgagct caagcttatg ggcatggcgc gaccacgggg cgcggggccc
      661 tgcagccccg ggttggagcg ggctccgcgc cggagcgtcg gggagctgcg cctgctcttc
      721 gaagcgggcg gatctggatc ggatccaccg gtcgccacca tggtgagcaa gggcgaggca
      781 gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag
      841 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      901 aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg
      961 tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc
     1021 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc
     1081 gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc
     1141 accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc
     1201 accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc
     1261 ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc
     1321 gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag
     1381 gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg
     1441 gacgagctgt accagggtgg cagcggtggc atggtgagca agggcgaggc agtgatcaag
     1501 gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc
     1561 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1621 aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc
     1681 agggccttca tcaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag
     1741 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag
     1801 gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc
     1861 cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg
     1921 ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac
     1981 ggcggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg
     2041 cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc
     2101 gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg
     2161 taccagggtg gcagcggtgg catggtgagc aagggcgagg cagtgatcaa ggagttcatg
     2221 cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     2281 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     2341 cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc cagggccttc
     2401 atcaagcacc ccgccgacat ccccgactac tataagcagt ccttccccga gggcttcaag
     2461 tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc
     2521 ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt ccctcctgac
     2581 ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc
     2641 gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc
     2701 tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat gcccggcgcc
     2761 tacaacgtcg accgcaagtt ggacatcacc tcccacaacg aggactacac cgtggtggaa
     2821 cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     2881 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2941 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3001 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3061 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3121 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3721 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4081 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4141 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4201 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4261 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4321 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4381 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4441 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4501 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4561 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4621 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4681 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4741 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4801 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4861 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4921 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4981 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5041 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5101 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5161 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5221 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5281 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5341 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5401 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5461 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5521 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5581 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5641 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5701 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5761 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5821 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5881 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5941 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6001 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6061 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6121 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6181 ctgattctgt ggataaccgt attaccgcca tgcat
//
