LOCUS       sequence-489857-        4726 bp    DNA     circular SYN 21-OCT-2025
DEFINITION  Expresses mNeonGreen-lifeactin fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   Zhang H, Lesnov GD, Subach OM, Zhang W, Kuzmicheva TP, Vlaskina AV, 
            Samygina VR, Chen L, Ye X, Nikolaeva AY, Gabdulkhakov A, Papadaki S,
            Qin W, Borshchevskiy V, Perfilov MM, Gavrikov AS, Drobizhev M, 
            Mishin AS, Piatkevich KD, Subach FV
  TITLE     Bright and stable monomeric green fluorescent protein derived from 
            StayGold.
  JOURNAL   Nat Methods. 2024 Apr;21(4):657-665. doi: 
            10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
  PUBMED    38409224
REFERENCE   2  (bases 1 to 4726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: mNeonGreen-lifeactin
FEATURES             Location/Qualifiers
     source          1..4726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      28..37
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             34..744
                     /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      289..298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    867..988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(904..923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     958..977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(995..1450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1132..1153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1344..1363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1477..1581
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1583..1940
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1583..1774
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1606..1626)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1791..1926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1975..2769
                     /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             1975..2769
                     /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(2029..2048)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2639..2658
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2957..2976)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3001..3048
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3377..3965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3866..3885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4140..4443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4444..4647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4598..4618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4676..4726
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
ORIGIN
        1 atccattcgt tgggggatcc accggtcgcc accatggtga gcaagggcga ggaggataac
       61 atggcctctc tcccagcgac acatgagtta cacatctttg gctccatcaa cggtgtggac
      121 tttgacatgg tgggtcaggg caccggcaat ccaaatgatg gttatgagga gttaaacctg
      181 aagtccacca agggtgacct ccagttctcc ccctggattc tggtccctca tatcgggtat
      241 ggcttccatc agtacctgcc ctaccctgac gggatgtcgc ctttccaggc cgccatggta
      301 gatggctccg gctaccaagt ccatcgcaca atgcagtttg aagatggtgc ctcccttact
      361 gttaactacc gctacaccta cgagggaagc cacatcaaag gagaggccca ggtgaagggg
      421 actggtttcc ctgctgacgg tcctgtgatg accaactcgc tgaccgctgc ggactggtgc
      481 aggtcgaaga agacttaccc caacgacaaa accatcatca gtacctttaa gtggagttac
      541 accacgggaa atggcaagcg ctaccggagc actgcgcgga ccacctacac ctttgccaag
      601 ccaatggcgg ctaactatct gaagaaccag ccgatgtacg tgttccgtaa gacggagctc
      661 aagcactcca agaccgagct caacttcaag gagtggcaaa aggcctttac cgatgtgatg
      721 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
      781 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
      841 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      961 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     1021 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1081 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     1141 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1201 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     1261 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     1321 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     1381 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     1441 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1501 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1561 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     1621 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1681 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1741 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     1801 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     1861 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     1921 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     1981 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2041 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2101 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     2161 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2221 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2281 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2341 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2401 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2461 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     2521 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2581 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     2641 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     2701 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     2761 ttcttctgag ctggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     2821 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     2881 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     2941 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3001 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     3061 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3121 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3181 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     3241 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3301 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3361 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3961 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4021 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     4081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     4141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4441 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     4501 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     4561 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     4621 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcatggg
     4681 cgtggccgac ctgatcaaga agttcgagag catcagcaag gaggag
//
