LOCUS       sequence-481796-        4736 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses Lifeact-mScarlet3-H (aka mYongHong) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4736)
  AUTHORS   Xiong H, Chang Q, Ding J, Wang S, Zhang W, Li Y, Wu Y, Lin P, Yang 
            C, Liu M, Fang G, Yang Y, Xie J, Qi D, Jiang T, Fu W, Hu F, Chen Y, 
            Yue R, Li Y, Cui Y, Li M, Fan S, Yang Y, Xu Y, Li D, Zhang F, Zhao 
            H, Wu C, Zheng Q, Piatkevich KD, Fu Z
  TITLE     A highly stable monomeric red fluorescent protein for advanced 
            microscopy.
  JOURNAL   Nat Methods. 2025 Apr 17. doi: 10.1038/s41592-025-02676-5.
  PUBMED    40247125
REFERENCE   2  (bases 1 to 4736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2025 Apr 17. doi: 10.1038/s41592-025-02676-5."
COMMENT     Sequence Label: pLifeact-mScarlet3-H (mYongHong)
FEATURES             Location/Qualifiers
     source          1..4736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        124..327
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     278..298
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             399..449
                     /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"
     regulatory      465..474
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             471..1157
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     869..888
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1283..1404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1320..1339)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1374..1393
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1411..1866)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1548..1569)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1760..1779
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1893..1997
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1999..2356
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1999..2190
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2022..2042)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2207..2342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2391..3185
                     /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             2391..3185
                     /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(2445..2464)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3055..3074
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3373..3392)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3417..3464
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3793..4381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4282..4301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(4556..4736,1..123)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
       61 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      121 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      181 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      241 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      301 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      361 cggactcaga tctcgagctc aagcttcgaa ttcccaccat gggtgtcgca gatttgatca
      421 agaaattcga aagcatctca aaggaagaac gggatccacc ggtcgccacc atggatagca
      481 ccgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg
      541 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
      601 ccaagctgag ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc
      661 agttcatgta cggctccagg gccttcacga agcaccccgc cgacatcccc gactactgga
      721 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
      781 ccgtgtccgt ggcccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc
      841 tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg
      901 aagcatccac cgagcggttg taccccgagg acgtcgtgct gaagggcgac attaagcatg
      961 ccctgcgcct gaaggacggc ggccggtacc tggcggactt caagaccacc tacagggcca
     1021 agaagcccgt gcagatgccc ggcgccttca acatcgaccg caagttggac atcacatccc
     1081 acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgtggcccgc cactccaccg
     1141 gcggctccgg tggctcctaa agcggccgcg actctagatc ataatcagcc ataccacatt
     1201 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1261 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1321 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1381 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     1441 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1501 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1561 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1621 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1681 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1741 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1801 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1861 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1921 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1981 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2101 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2161 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2221 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2281 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2341 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     2401 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2461 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2521 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     2581 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2641 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2701 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2761 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2821 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2881 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     2941 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3001 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3061 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3121 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3181 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3241 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3301 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     3361 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     3421 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     3481 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3541 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3601 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     3661 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3721 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4441 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     4501 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4561 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4621 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4681 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgcca
//
