LOCUS       sequence-481795-        5014 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses H2B-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  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 5014)
  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: pH2B-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      214..802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     703..722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        977..1280
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1281..1484
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1435..1455
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1900..1909
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1906..2592
                     /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     2304..2323
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2718..2839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2755..2774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2809..2828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2846..3301)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2983..3004)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3195..3214
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3328..3432
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3434..3791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3434..3625
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3457..3477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3642..3777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3826..4620
                     /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             3826..4620
                     /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(3880..3899)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4490..4509
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4808..4827)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4852..4899
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
       61 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
      121 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      841 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      901 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      961 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     1021 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     1081 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     1141 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     1201 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     1261 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     1321 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     1381 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     1441 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     1501 agatccgcta gcatgcctga accggcaaaa tccgctccgg cccctaaaaa gggctccaag
     1561 aaagccgtca ccaaagccca gaagaaagac ggcaagaagc gcaagcgcag ccgcaaagag
     1621 agctactcca tctacgtgta caaggtgctg aagcaggtcc accccgacac cggcatctcg
     1681 tccaaggcca tgggcatcat gaactccttc gtcaacgaca tcttcgagcg catcgcggga
     1741 gaggcttccc gcctggcgca ctacaacaag cgctccacca tcacatcccg cgagatccag
     1801 acggccgtgc gcctgctgct gcccggcgag ctggccaagc acgccgtgtc cgagggcacc
     1861 aaggcggtca ccaagtacac cagctccaag ccaccggtcg ccaccatgga tagcaccgag
     1921 gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac
     1981 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2041 ctgagggtga ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc
     2101 atgtacggct ccagggcctt cacgaagcac cccgccgaca tccccgacta ctggaagcag
     2161 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg
     2221 tccgtggccc aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc
     2281 ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagca
     2341 tccaccgagc ggttgtaccc cgaggacgtc gtgctgaagg gcgacattaa gcatgccctg
     2401 cgcctgaagg acggcggccg gtacctggcg gacttcaaga ccacctacag ggccaagaag
     2461 cccgtgcaga tgcccggcgc cttcaacatc gaccgcaagt tggacatcac atcccacaac
     2521 gaggactaca ccgtggtgga acagtacgaa cgctccgtgg cccgccactc caccggcggc
     2581 tccggtggct cctaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2641 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2701 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2761 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2821 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2881 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2941 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3001 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3061 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3121 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3181 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3241 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3301 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3361 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3421 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3661 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3721 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3781 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4321 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4381 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4441 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4561 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4621 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4681 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4741 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4801 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4861 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4921 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4981 cttccttttc cccaccccac cccccaagtt cggg
//
