LOCUS       sequence-481790-        5095 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses Tom20-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5095)
  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 5095)
  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: pTom20-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..5095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        40..231
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(63..83)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      248..383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             432..1226
                     /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             432..1226
                     /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(486..505)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1096..1115
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1414..1433)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1458..1505
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1834..2422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2323..2342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2597..2900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2901..3104
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3055..3075
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3133..3142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3598..3607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3604..4290
                     /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     4002..4021
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4419..4540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4456..4475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4510..4529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4547..5002)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4684..4705)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4896..4915
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5029..5095,1..38)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
       61 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      121 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      241 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
      301 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
      361 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
      421 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      481 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      541 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      601 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      661 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      721 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      781 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      841 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
      901 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
      961 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1021 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1081 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1141 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1201 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1261 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1321 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1381 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     1441 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     1501 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     1561 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     1621 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     1681 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1741 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1801 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1861 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1921 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1981 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2341 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2401 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2461 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2521 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     2581 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2641 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2701 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2761 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2821 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     2881 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     2941 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     3001 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     3061 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     3121 agatccgcta gcgccaccat ggtgggtcgg aacagcgcca tcgccgccgg tgtatgcggg
     3181 gcccttttca ttgggtactg catctacttc gaccgcaaaa gacgaagtga ccccaacttc
     3241 aagaacaggc ttcgagaacg aagaaagaaa cagaagcttg ccaaggagag agctgggctt
     3301 tccaagttac ctgaccttaa agatgctgaa gctgttcaga agttcttcct tgaagaaata
     3361 cagcttggtg aagagttact agctcaaggt gaatatgaga agggcgtaga ccatctgaca
     3421 aatgcaattg ctgtgtgtgg acagccacag cagttactgc aggtcttaca gcaaactctt
     3481 ccaccaccag tgttccagat gcttctgact aagctcccaa caattagtca gagaattgta
     3541 agtgctcaga gcttggctga agatgatgtg gaaggcggta gcggggatcc accggtcgcc
     3601 accatggata gcaccgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
     3661 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     3721 ggcacccaga ccgccaagct gagggtgacc aagggtggcc ccctgccctt ctcctgggac
     3781 atcctgtccc ctcagttcat gtacggctcc agggccttca cgaagcaccc cgccgacatc
     3841 cccgactact ggaagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     3901 gaggacggcg gcgccgtgtc cgtggcccag gacacctccc tggaggacgg caccctgatc
     3961 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
     4021 acaatgggct gggaagcatc caccgagcgg ttgtaccccg aggacgtcgt gctgaagggc
     4081 gacattaagc atgccctgcg cctgaaggac ggcggccggt acctggcgga cttcaagacc
     4141 acctacaggg ccaagaagcc cgtgcagatg cccggcgcct tcaacatcga ccgcaagttg
     4201 gacatcacat cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgtggcc
     4261 cgccactcca ccggcggctc cggtggctcc taataaagcg gccgcgactc tagatcataa
     4321 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4381 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4441 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4501 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     4561 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4621 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4681 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4741 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4801 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4861 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4921 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4981 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5041 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataac
//
