LOCUS       sequence-482768-        4822 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Expresses Cox8a-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4822)
  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 4822)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 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: pCox8a-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..4822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..115
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             173..247
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             269..340
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      368..377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             374..1060
                     /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     772..791
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1186..1307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1223..1242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1277..1296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1314..1769)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1451..1472)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1663..1682
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1796..1900
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1902..2259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1902..2093
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1925..1945)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2110..2245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2294..3088
                     /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             2294..3088
                     /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(2348..2367)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2958..2977
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3276..3295)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3320..3367
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3696..4284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4185..4204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4459..4762
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4763..4822,1..144)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
       61 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      121 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatgtccgt
      181 cctgacgccg ctgctgctgc ggggcttgac aggctcggcc cggcggctcc cagtgccgcg
      241 cgccaagatc cattcgttgg gggatctgtc cgtcctgacg ccgctgctgc tgcggggctt
      301 gacaggctcg gcccggcggc tcccagtgcc gcgcgccaag atccattcgt tgggggatcc
      361 accggtcgcc accatggata gcaccgaggc agtgatcaag gagttcatgc ggttcaaggt
      421 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
      481 cccctacgag ggcacccaga ccgccaagct gagggtgacc aagggtggcc ccctgccctt
      541 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca cgaagcaccc
      601 cgccgacatc cccgactact ggaagcagtc cttccccgag ggcttcaagt gggagcgcgt
      661 gatgaacttc gaggacggcg gcgccgtgtc cgtggcccag gacacctccc tggaggacgg
      721 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
      781 gcagaagaag acaatgggct gggaagcatc caccgagcgg ttgtaccccg aggacgtcgt
      841 gctgaagggc gacattaagc atgccctgcg cctgaaggac ggcggccggt acctggcgga
      901 cttcaagacc acctacaggg ccaagaagcc cgtgcagatg cccggcgcct tcaacatcga
      961 ccgcaagttg gacatcacat cccacaacga ggactacacc gtggtggaac agtacgaacg
     1021 ctccgtggcc cgccactcca ccggcggctc cggtggctcc taaagcggcc gcgactctag
     1081 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1141 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1201 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1261 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     1321 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1381 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1441 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1501 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1561 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1621 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1681 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     1741 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     1801 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1861 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     1921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2161 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2281 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2341 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2401 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2461 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2521 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2581 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2641 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2701 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2761 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2821 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2881 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2941 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3001 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3061 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3121 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3181 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3241 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3301 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3361 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3421 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3481 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3541 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3841 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4321 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4381 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     4441 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     4501 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     4561 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     4621 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     4681 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4741 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     4801 gcggtttgac tcacggggat tt
//
