LOCUS       sequence-481808-        4743 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses mScarlet3-H (aka mYongHong) localized to ER in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  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 4743)
  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: pKDEL-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..170
                     /gene="bla"
                     /label=AmpR promoter
     promoter        172..529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        172..363
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(195..215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      380..515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             564..1358
                     /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             564..1358
                     /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(618..637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1228..1247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1546..1565)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1590..1637
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1966..2554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2455..2474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2729..3032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3033..3236
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3187..3207
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3325..3334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3331..4017
                     /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     3729..3748
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4199..4320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4236..4255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4290..4309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4327..4743,1..39))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4464..4485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4676..4695
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
       61 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      121 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
      181 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
      241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
      481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
      541 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
      601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
      661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
      721 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
      781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
      841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
      901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
      961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     1021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     1081 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     1141 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     1201 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     1261 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     1321 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     1381 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     1441 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     1501 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     1561 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     1621 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     1681 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     1741 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     1801 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtatc
     2161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2641 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     2701 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     2761 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     2821 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     2881 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     2941 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     3001 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     3061 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     3121 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     3181 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     3241 tagtgaaccg tcagatccgc tagcatgctg ctatccgtgc cgttgctgct cggcctcctc
     3301 ggcctggccg tcgccgcacc ggtcgccacc atggatagca ccgaggcagt gatcaaggag
     3361 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
     3421 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgag ggtgaccaag
     3481 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
     3541 gccttcacga agcaccccgc cgacatcccc gactactgga agcagtcctt ccccgagggc
     3601 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgtccgt ggcccaggac
     3661 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     3721 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcatccac cgagcggttg
     3781 taccccgagg acgtcgtgct gaagggcgac attaagcatg ccctgcgcct gaaggacggc
     3841 ggccggtacc tggcggactt caagaccacc tacagggcca agaagcccgt gcagatgccc
     3901 ggcgccttca acatcgaccg caagttggac atcacatccc acaacgagga ctacaccgtg
     3961 gtggaacagt acgaacgctc cgtggcccgc cactccaccg gcggctccgg tggctcctcc
     4021 ggaaaggacg agctgtaaga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
     4081 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4141 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4321 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4381 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4441 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4501 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4561 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4621 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4681 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4741 cca
//
