LOCUS       sequence-261572-        6950 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 6950)
  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 6950)
  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: 3xmNeonGreen_UtrCH
FEATURES             Location/Qualifiers
     source          1..6950
                     /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..1317
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1336..2040
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1591..1600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2059..2766
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2314..2323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3562..3614
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3738..3859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3775..3794)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3829..3848
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3866..4321)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4003..4024)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4215..4234
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4348..4452
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4454..4811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4454..4645
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4477..4497)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4662..4797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4846..5640
                     /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             4846..5640
                     /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(4900..4919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5510..5529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5828..5847)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5872..5919
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6248..6836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6737..6756
                     /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 gaggataaca tggcctctct cccagcgaca
      661 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      721 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      781 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
      841 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ctaccaagtc
      901 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
      961 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1021 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1081 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtaccag
     1321 ggtggcagcg gtggcatggt gagcaagggc gaggaggata acatggcctc tctcccagcg
     1381 acacatgagt tacacatctt tggctccatc aacggtgtgg actttgacat ggtgggtcag
     1441 ggcaccggca atccaaatga tggttatgag gagttaaacc tgaagtccac caagggtgac
     1501 ctccagttct ccccctggat tctggtccct catatcgggt atggcttcca tcagtacctg
     1561 ccctaccctg acgggatgtc gcctttccag gccgccatgg tagatggctc cggctaccaa
     1621 gtccatcgca caatgcagtt tgaagatggt gcctccctta ctgttaacta ccgctacacc
     1681 tacgagggaa gccacatcaa aggagaggcc caggtgaagg ggactggttt ccctgctgac
     1741 ggtcctgtga tgaccaactc gctgaccgct gcggactggt gcaggtcgaa gaagacttac
     1801 cccaacgaca aaaccatcat cagtaccttt aagtggagtt acaccactgg aaatggcaag
     1861 cgctaccgga gcactgcgcg gaccacctac acctttgcca agccaatggc ggctaactat
     1921 ctgaagaacc agccgatgta cgtgttccgt aagacggagc tcaagcactc caagaccgag
     1981 ctcaacttca aggagtggca aaaggccttt accgatgtga tgggcatgga cgagctgtac
     2041 cagggtggca gcggtggcat ggtgagcaag ggcgaggagg ataacatggc ctctctccca
     2101 gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt
     2161 cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt
     2221 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
     2281 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggctac
     2341 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     2401 acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct
     2461 gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact
     2521 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc
     2581 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     2641 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc
     2701 gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg
     2761 tacaagtccg gaaccatggc caagtatgga gaacatgaag ccagtcctga caatgggcag
     2821 aacgaattca gtgatatcat taagtccaga tctgatgaac acaatgacgt acagaagaaa
     2881 acctttacca aatggataaa tgctcgattt tcaaagagtg ggaaaccacc catcaatgat
     2941 atgttcacag acctcaaaga tggaaggaag ctattggatc ttctagaagg cctcacagga
     3001 acatcactgc caaaggaacg tggttccaca agggtacatg ccttaaataa cgtcaacaga
     3061 gtgctgcagg ttttacatca gaacaatgtg gaattagtga atataggggg aaccgacatt
     3121 gtggatggaa atcacaaact gactttgggg ttactttgga gcatcatttt gcactggcag
     3181 gtgaaagatg tcatgaagga tgtcatgtcg gacctgcagc agacgaacag tgagaagatc
     3241 ctgctcagct gggtgcgtca gaccaccagg ccctacagcc aagtcaacgt cctcaacttc
     3301 accaccagct ggacagatgg actcgccttt aatgctgtcc tccaccgaca taaacctgat
     3361 ctcttcagct gggataaagt tgtcaaaatg tcaccaattg agagacttga acatgccttc
     3421 agcaaggctc aaacttattt gggaattgaa aagctgttag atcctgaaga tgttgccgtt
     3481 cggcttcctg acaagaaatc cataattatg tatttaacat ctttgtttga ggtgctacct
     3541 cagcaagtca ccatagacta actcgagctc aagcttcgaa ttctgcagtc gacggtaccg
     3601 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3661 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3721 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3781 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3841 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3901 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3961 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4021 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4081 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4141 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4201 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4261 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4321 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4381 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4441 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4501 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4561 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4621 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4681 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4741 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4801 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4861 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4921 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4981 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5041 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5101 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5161 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5221 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5281 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5341 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5401 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5461 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5521 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5581 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5641 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5701 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5761 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5821 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5881 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5941 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6001 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6061 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6121 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6181 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6241 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6301 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6361 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6421 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6481 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6541 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6601 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6661 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6721 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6781 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6841 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6901 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
