LOCUS       sequence-481794-        4824 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses phb2-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4824)
  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 4824)
  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: pPhb2-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..4824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     150..169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        424..727
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        728..931
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     882..902
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1155..1164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1161..1847
                     /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     1559..1578
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1975..2096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2012..2031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2066..2085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2103..2558)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2240..2261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2452..2471
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2585..2689
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2691..3048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2691..2882
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2714..2734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2899..3034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3083..3877
                     /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             3083..3877
                     /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(3137..3156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3747..3766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4065..4084)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4109..4156
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4485..4824,1..249)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
       61 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
      361 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      421 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      481 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      541 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      601 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      661 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      721 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      781 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      841 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      901 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagca
      961 tggcccagaa cttgaaggac ttggcgggac ggctgcccgc cgggccccgg ggcatgggca
     1021 cggccctgaa gctgttgctg ggggccggcg ccgtggccta cggtgtgcgc gaatctgtgt
     1081 tcaccgtgga aggcgggcac agagccatcg gaggaagtgg aggcagggta ccgcgggccc
     1141 gggatccacc ggtcgccacc atggatagca ccgaggcagt gatcaaggag ttcatgcggt
     1201 tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1261 agggccgccc ctacgagggc acccagaccg ccaagctgag ggtgaccaag ggtggccccc
     1321 tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcacga
     1381 agcaccccgc cgacatcccc gactactgga agcagtcctt ccccgagggc ttcaagtggg
     1441 agcgcgtgat gaacttcgag gacggcggcg ccgtgtccgt ggcccaggac acctccctgg
     1501 aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct cctgacggcc
     1561 ccgtaatgca gaagaagaca atgggctggg aagcatccac cgagcggttg taccccgagg
     1621 acgtcgtgct gaagggcgac attaagcatg ccctgcgcct gaaggacggc ggccggtacc
     1681 tggcggactt caagaccacc tacagggcca agaagcccgt gcagatgccc ggcgccttca
     1741 acatcgaccg caagttggac atcacatccc acaacgagga ctacaccgtg gtggaacagt
     1801 acgaacgctc cgtggcccgc cactccaccg gcggctccgg tggctcctaa taagcggccg
     1861 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1921 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1981 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2041 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2101 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2161 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2221 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2281 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2341 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2401 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2461 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2521 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2581 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2641 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2701 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2761 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2821 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2881 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2941 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3001 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3061 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3121 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3181 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3241 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3301 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3361 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3421 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3481 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3541 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3601 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3661 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3721 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3781 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3841 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3901 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3961 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4021 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4081 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4141 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4201 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4261 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4321 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4381 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4621 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4801 tgaacggggg gttcgtgcac acag
//
