LOCUS       sequence-482764-        4782 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Expresses CytERM-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4782)
  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 4782)
  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: pCytERM-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..4782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..160
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             209..1003
                     /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             209..1003
                     /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(263..282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     873..892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1191..1210)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1235..1282
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1611..2199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2100..2119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2374..2677
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2678..2881
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2832..2852
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3065..3074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3071..3757
                     /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     3469..3488
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3883..4004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3920..3939)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3974..3993
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4011..4466)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4148..4169)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4360..4379
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4493..4597
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4599..4782,1..174)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(4599..4782,1..8)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4622..4642)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
       61 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
      121 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
      181 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
      241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      361 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
      421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
      541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
      601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
      661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
      721 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
      781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
      841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
      901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
      961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1021 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     1081 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     1141 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     1201 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     1261 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     1321 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     1381 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     1441 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     1501 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1561 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1621 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1681 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1741 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1801 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1861 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1921 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1981 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2041 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2101 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2161 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2221 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2281 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     2341 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2401 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2461 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2521 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2581 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2641 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2701 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2761 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2821 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2881 tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcca ccatggatcc
     2941 tgtggtggtg ctggggctct gtctctcctg tttgcttctc ctttcactct ggaaacagag
     3001 ctatggggga gggaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc
     3061 ggtcgccacc atggatagca ccgaggcagt gatcaaggag ttcatgcggt tcaaggtgca
     3121 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     3181 ctacgagggc acccagaccg ccaagctgag ggtgaccaag ggtggccccc tgcccttctc
     3241 ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcacga agcaccccgc
     3301 cgacatcccc gactactgga agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3361 gaacttcgag gacggcggcg ccgtgtccgt ggcccaggac acctccctgg aggacggcac
     3421 cctgatctac aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca
     3481 gaagaagaca atgggctggg aagcatccac cgagcggttg taccccgagg acgtcgtgct
     3541 gaagggcgac attaagcatg ccctgcgcct gaaggacggc ggccggtacc tggcggactt
     3601 caagaccacc tacagggcca agaagcccgt gcagatgccc ggcgccttca acatcgaccg
     3661 caagttggac atcacatccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc
     3721 cgtggcccgc cactccaccg gcggctccgg tggctcctaa agcggccgcg actctagatc
     3781 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3841 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3901 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3961 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     4021 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4081 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4141 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4201 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4261 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4321 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4381 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4441 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4501 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4561 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4621 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4681 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4741 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa cc
//
