LOCUS       sequence-481797-        5029 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses Giantin-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5029)
  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 5029)
  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: pGiantin-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..5029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     262..281
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1084..1205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1121..1140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1175..1194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1212..1667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1349..1370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1561..1580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1694..1798
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1800..2157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1800..1991
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1823..1843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2008..2143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2192..2986
                     /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             2192..2986
                     /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(2246..2265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2856..2875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3174..3193)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3218..3265
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3594..4182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4083..4102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4357..4660
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4661..4864
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4815..4835
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(4893..5029,1..550)
                     /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"
ORIGIN
        1 gagggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat
       61 gtacggctcc agggccttca cgaagcaccc cgccgacatc cccgactact ggaagcagtc
      121 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgtc
      181 cgtggcccag gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg
      241 caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcatc
      301 caccgagcgg ttgtaccccg aggacgtcgt gctgaagggc gacattaagc atgccctgcg
      361 cctgaaggac ggcggccggt acctggcgga cttcaagacc acctacaggg ccaagaagcc
      421 cgtgcagatg cccggcgcct tcaacatcga ccgcaagttg gacatcacat cccacaacga
      481 ggactacacc gtggtggaac agtacgaacg ctccgtggcc cgccactcca ccggcggctc
      541 cggtggctcc tccggactca gatctcgagg agaaccgcag caaagctttt ctgaagctca
      601 gcagcagcta tgcaacacca gacaggaagt gaatgaatta aggaagctgc tggaagaaga
      661 acgagaccaa agagtggctg ctgagaatgc tctctctgtg gccgaggagc agatcagacg
      721 gttagagcac agtgaatggg actcttcccg gactcctatc attggctcct gtggcactca
      781 ggagcaggca ctgttaatag atcttacaag caacagttgt cgaaggaccc ggagtggcgt
      841 tggatggaag cgagtcctgc gttcactctg tcattcacgg acccgagtgc cacttctagc
      901 agccatctac tttctaatga ttcatgtcct gctcattctg tgttttacgg gccatctata
      961 gagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     1021 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1081 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1141 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1201 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1261 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1321 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1381 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1441 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1501 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1561 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1621 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     1681 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1741 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     1801 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1861 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     1921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1981 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2041 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2101 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     2161 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2221 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2281 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2341 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     2401 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2461 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2521 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2581 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2641 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2701 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2761 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2821 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2881 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2941 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3001 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3061 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3121 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     3181 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3241 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3301 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3361 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3421 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     3481 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3541 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3601 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3661 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3721 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3781 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3841 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3901 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3961 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4021 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4081 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4141 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4201 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4261 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     4321 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     4381 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     4441 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     4501 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     4561 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4621 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     4681 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4741 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4801 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4861 agctggttta gtgaaccgtc agatccgcta gcatggatag caccgaggca gtgatcaagg
     4921 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     4981 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagct
//
