LOCUS       sequence-481802-        5522 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses EMTB-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  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 5522)
  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: pEMTB-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..5522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     493..512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        767..1070
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1071..1274
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1225..1245
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1341..1350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1350..1379
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2199..2208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2205..2891
                     /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     2603..2622
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3016..3137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3053..3072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3107..3126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3144..3599)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3281..3302)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3493..3512
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3626..3730
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3732..4089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3732..3923
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3755..3775)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3940..4075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4124..4918
                     /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             4124..4918
                     /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(4178..4197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4788..4807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5106..5125)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5150..5197
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
       61 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      661 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
      721 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      781 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      841 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      901 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      961 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1021 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     1081 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     1141 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1201 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1261 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
     1321 cgagctcaag cttcgaattc gccaccatgg agcagaagct catctcagaa gaagacctcg
     1381 gcgcgccagt gcgaagcgaa acagcacccg acagctacaa agtgcaagat aagaaaaatg
     1441 cctccagccg ccctgcctct gcaatttcag gacaaaataa caaccactca ggaaataaac
     1501 cagaccctcc gcctgtgtta cgtgttgatg accggcagcg gctggcccgg gagcgacgtg
     1561 aggaacggga gaaacagcta gctgcaagag aaatagtgtg gttagaaaga gaagagcgag
     1621 ccaggcagca ctacgagaag cacctggaag agcggaagaa gaggttggag gagcagaggc
     1681 agaaggagga gcggaggagg gctgctgtgg aggagaagcg gaggcagaga cttgaggagg
     1741 acaaagaacg ccacgaagct gttgtacggc gcacaatgga aaggagccag aagccaaaac
     1801 agaagcataa ccgttggtcg tggggaggct ctctccatgg gagccctagc atccacagtg
     1861 cagatccaga caggcggtca gtttccacca tgaatctttc gaaatatgtt gatcccgtca
     1921 ttagcaagcg gctctcctct tcatctgcaa ctttactaaa ttctccagat agagctcgcc
     1981 gcctgcagct cagcccatgg gagagcagcg ttgttaacag actcctgacg cccacacatt
     2041 cgttcctggc cagaagtaaa agcacagctg ccttgtctgg agaagcagca tcttgcagcc
     2101 ccatcatcat gccctacaaa gctgcacact ctagaaattc gatggatcga ccaaaactct
     2161 ttgtaacacc acctgagggc tcttcggatc caccggtcgc caccatggat agcaccgagg
     2221 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg
     2281 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     2341 tgagggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
     2401 tgtacggctc cagggccttc acgaagcacc ccgccgacat ccccgactac tggaagcagt
     2461 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtgt
     2521 ccgtggccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg
     2581 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcat
     2641 ccaccgagcg gttgtacccc gaggacgtcg tgctgaaggg cgacattaag catgccctgc
     2701 gcctgaagga cggcggccgg tacctggcgg acttcaagac cacctacagg gccaagaagc
     2761 ccgtgcagat gcccggcgcc ttcaacatcg accgcaagtt ggacatcaca tcccacaacg
     2821 aggactacac cgtggtggaa cagtacgaac gctccgtggc ccgccactcc accggcggct
     2881 ccggtggctc ctaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     2941 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3001 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3061 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3121 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3181 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3241 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3301 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3361 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3421 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3481 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3541 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3601 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3661 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3721 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3901 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3961 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4021 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4081 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4141 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4201 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4261 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4321 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4381 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4441 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4501 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4561 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4621 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4681 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4741 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4801 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4861 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4921 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4981 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5041 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5101 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5161 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5221 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5281 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5341 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tc
//
