LOCUS       sequence-481809-        4924 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses Sec61b-mScarlet3-H in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4924)
  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 4924)
  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: pSec61-beta-mScarlet3-H (aka mYongHong)
FEATURES             Location/Qualifiers
     source          1..4924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      95..683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     584..603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        858..1161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1162..1365
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1316..1336
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1394..2080
                     /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     1792..1811
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2096..2386
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    2509..2630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2546..2565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2600..2619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2637..3092)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2774..2795)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2986..3005
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3119..3223
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3225..3582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3225..3416
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3248..3268)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3433..3568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3617..4411
                     /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             3617..4411
                     /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(3671..3690)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4281..4300
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4599..4618)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4643..4690
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
       61 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      121 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      181 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      241 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      301 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      361 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      421 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      481 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      541 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      601 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      661 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      721 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      781 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
      841 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      901 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      961 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     1021 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     1081 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     1141 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     1201 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     1261 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     1321 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     1381 cagatccgct agcatggata gcaccgaggc agtgatcaag gagttcatgc ggttcaaggt
     1441 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1501 cccctacgag ggcacccaga ccgccaagct gagggtgacc aagggtggcc ccctgccctt
     1561 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca cgaagcaccc
     1621 cgccgacatc cccgactact ggaagcagtc cttccccgag ggcttcaagt gggagcgcgt
     1681 gatgaacttc gaggacggcg gcgccgtgtc cgtggcccag gacacctccc tggaggacgg
     1741 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
     1801 gcagaagaag acaatgggct gggaagcatc caccgagcgg ttgtaccccg aggacgtcgt
     1861 gctgaagggc gacattaagc atgccctgcg cctgaaggac ggcggccggt acctggcgga
     1921 cttcaagacc acctacaggg ccaagaagcc cgtgcagatg cccggcgcct tcaacatcga
     1981 ccgcaagttg gacatcacat cccacaacga ggactacacc gtggtggaac agtacgaacg
     2041 ctccgtggcc cgccactcca ccggcggctc cggtggctcc ccaccggtcg ccaccatgcc
     2101 tggtccgacc cccagtggca ctaacgtggg atcctcaggg cgctctccca gcaaagcagt
     2161 ggccgcccgg gcggcgggat ccactgtccg gcagaggaaa aatgccagct gtgggacaag
     2221 gagtgcaggc cgcacaacct cggcaggcac cggggggatg tggcgattct acacagaaga
     2281 ttcacctggg ctcaaagttg gccctgttcc agtattggtt atgagtcttc tgttcatcgc
     2341 ttctgtattt atgttgcaca tttggggcaa gtacactcgt tcgtagagcg gccgcgactc
     2401 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2461 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2521 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2581 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     2641 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2701 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2761 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2821 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2881 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2941 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3001 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3061 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3121 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3181 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3241 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3301 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3361 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3421 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3481 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3541 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3601 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3661 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3721 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3781 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3841 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3901 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3961 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4021 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4081 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4141 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4201 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4261 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4321 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4381 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4441 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4501 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4561 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4621 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4681 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4741 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4801 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4861 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4921 gtga
//
