LOCUS       sequence-481791-        6024 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses Plin5-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6024)
  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 6024)
  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: pPlin5-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..6024
                     /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"
     regulatory      1754..1763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1760..2446
                     /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     2158..2177
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2571..2692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2608..2627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2662..2681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2699..3154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2836..2857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3048..3067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3181..3285
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3287..3644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3287..3478
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3310..3330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3495..3630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3679..4473
                     /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             3679..4473
                     /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(3733..3752)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4343..4362
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4661..4680)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4705..4752
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5081..5669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5570..5589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5844..6024,1..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 ctagcatgag
      361 cgaggaggag gctgctcaga tccccagaag cagcgtgtgg gagcaggacc agcagaacgt
      421 ggtgcagaga gtggtggccc tgcctctggt gagagctaca tgcaccgccg tgtgcgacgt
      481 gtacagcgct gctaaagaca gacaccccct gctgggcagc gcttgtagac tggctgagaa
      541 ttgcgtgtgc ggcctgacca ccagagctct ggatcacgct cagcccctgc tggaacacct
      601 gcagcctcag ctggctacca tgaattccct ggcttgcaga ggcctggaca agctggaaga
      661 gaaactgccc ttcctgcagc agccctccga gacagtggtg acaagcgcta aagacgtggt
      721 ggcctcctcc gtgaccggag tggtggatct ggctagaaga ggcagaagat ggagcgtgga
      781 actgaagagg agcgtgagcc acgccgtgga tgtggtgctg gaaaagagcg aggagctggt
      841 ggaccatttt ctgcccatga ctgaggaaga gctggccgct ctggccgctg aagctgaagg
      901 acctgaggtg ggaagcgtgg aagaccagag aagacagcag ggatacttcg tgagactggg
      961 atccctgagc gccagaatca gacacctggc ctacgagcac tccgtgggaa aactgagaca
     1021 gagcaagcac agagcccagg acaccctggc tcagctgcag gaaaccctgg agctgattga
     1081 ccacatgcag tgcggcgtga cccctacagc tcctgcttgt cctggcaaag tgcacgagct
     1141 gtggggcgaa tggggccaga gacctcctga aagcagaaga agaagccagg ccgagctgga
     1201 gaccctggtg ctgtctagat ccctgactca ggagctgcag ggaacagtgg aggccctgga
     1261 aagcagcgtg agaggactgc cagccggagc tcaggaaaag gtggctgagg tgagaagaag
     1321 cgtggatgcc ctgcagaccg ccttcgctga tgctagatgc ttcagagacg tgcccgccgc
     1381 tgctctggct gaaggaagag gaagagtggc ccacgcccat gcttgtgtgg acgaactgct
     1441 ggagctggtg gtgcaggctg tgcctctgcc ttggctggtg ggacctttcg ctcctattct
     1501 ggtggagagg cccgagcctc tgcctgatct ggctgatctg gtggatgaag tgatcggcgg
     1561 ccctgatccc agatgggctc atctggactg gcctgctcag cagagggctt gggaagctga
     1621 gcacagagac ggatccggca acggagacgg agacagaatg ggagtggccg gagatatttg
     1681 cgagcaggag cctgaaacac ccagctgccc agtgaagcac accctgatgc ccgaactgga
     1741 cttcccaccg gtcgccacca tggatagcac cgaggcagtg atcaaggagt tcatgcggtt
     1801 caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     1861 gggccgcccc tacgagggca cccagaccgc caagctgagg gtgaccaagg gtggccccct
     1921 gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcacgaa
     1981 gcaccccgcc gacatccccg actactggaa gcagtccttc cccgagggct tcaagtggga
     2041 gcgcgtgatg aacttcgagg acggcggcgc cgtgtccgtg gcccaggaca cctccctgga
     2101 ggacggcacc ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc
     2161 cgtaatgcag aagaagacaa tgggctggga agcatccacc gagcggttgt accccgagga
     2221 cgtcgtgctg aagggcgaca ttaagcatgc cctgcgcctg aaggacggcg gccggtacct
     2281 ggcggacttc aagaccacct acagggccaa gaagcccgtg cagatgcccg gcgccttcaa
     2341 catcgaccgc aagttggaca tcacatccca caacgaggac tacaccgtgg tggaacagta
     2401 cgaacgctcc gtggcccgcc actccaccgg cggctccggt ggctcctaaa gcggcccgac
     2461 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2521 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2581 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2641 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     2701 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2761 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2821 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2881 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2941 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3001 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3061 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3121 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3181 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3241 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3301 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3361 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3421 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3481 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3541 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3601 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3661 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3721 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3781 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3841 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3901 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3961 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4021 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4081 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4141 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4201 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4261 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4321 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4381 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4441 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4501 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4561 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4621 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4681 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4741 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4801 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4861 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4921 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4981 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5041 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5101 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5161 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5221 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5281 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5341 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5401 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5461 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5521 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5581 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5641 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5701 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5761 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     5821 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5881 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5941 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6001 gtacatcaag tgtatcatat gcca
//
