LOCUS       sequence-495653-        4964 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Expression of mScarelt3-S2 fused with Sec61-beta. This construct is 
            used for imaging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4964)
  AUTHORS   Ding Y, He W, Wang K, Xue F, Zheng K, Zhao S, Qi D, Li T, Xia P, Pan
            L, Chen L, Chen Z, Yuan L, Xu P
  TITLE     A highly photostable monomeric red fluorescent protein for 
            dual-color 3D STED and time-lapse 3D SIM imaging.
  JOURNAL   Nat Methods. 2025 Nov 26. doi: 10.1038/s41592-025-02962-2.
  PUBMED    41299071
REFERENCE   2  (bases 1 to 4964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2025 Nov 26. doi: 10.1038/s41592-025-02962-2."
COMMENT     Sequence Label: mScarlet3-S2-Sec61-beta
FEATURES             Location/Qualifiers
     source          1..4964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             600..1286
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFQYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     998..1017
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1314..1601
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="PGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAGR
                     TTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    1752..1873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1789..1808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1843..1862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1880..2335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2017..2038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2229..2248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2362..2466
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2468..2825
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2468..2659
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2491..2511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2676..2811
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2860..3654
                     /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             2860..3654
                     /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(2914..2933)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3524..3543
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3842..3861)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3886..3933
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4262..4850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4751..4770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 tggatagcac cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct
      661 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      721 cccagaccgc caagctgagg gtgaccaagg gtggccccct gcccttctcc tgggacatcc
      781 tgtcccctca gttccagtac ggctccaggg ccttcacgaa gcaccccgcc gacatccccg
      841 actactggaa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      901 acggcggcgc cgtgtccgtg gcccaggaca cctccctgga ggacggcacc ctgatctaca
      961 aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa
     1021 tgggctggga agcatccacc gagcggttgt accccgagga cgtcgtgctg aagggcgaca
     1081 ttaagcacgc cctgcgcctg aaggacggcg gccgctacct ggcggacttc aagaccacct
     1141 acagggccaa gaagcccgtg cagatgcccg gcgccttcaa catcgaccgc aagttggaca
     1201 tcacatccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc gtggcccgcc
     1261 actccaccgg cggctccggt ggctcccaga tctcgagctc aagcttcgaa ttccctggtc
     1321 cgacccccag tggcactaac gtgggatcct cagggcgctc tcccagcaaa gcagtggccg
     1381 cccgggcggc gggatccact gtccggcaga ggaaaaatgc cagctgtggg acaaggagtg
     1441 caggccgcac aacctcggca ggcaccgggg ggatgtggcg attctacaca gaagattcac
     1501 ctgggctcaa agttggccct gttccagtat tggttatgag tcttctgttc atcgcttctg
     1561 tatttatgtt gcacatttgg ggcaagtaca ctcgttcgta ggtcgacggt accgcgggcc
     1621 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     1681 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     1741 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1861 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     1921 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1981 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2101 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     2341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2461 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2521 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2581 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2641 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2701 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2761 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2821 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     2881 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2941 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3001 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     3061 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3121 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3181 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3241 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3301 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3361 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     3421 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3481 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3541 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3601 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3661 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     3721 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     3781 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     3841 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     3901 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     3961 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4021 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4081 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     4141 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4381 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4921 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
