LOCUS       sequence-263632-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ER-targeted Ribosome non-stalling reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  AUTHORS   Wang L, Xu Y, Rogers H, Saidi L, Noguchi CT, Li H, Yewdell JW, 
            Guydosh NR, Ye Y
  TITLE     UFMylation of RPL26 links translocation-associated quality control 
            to endoplasmic reticulum protein homeostasis.
  JOURNAL   Cell Res. 2020 Jan;30(1):5-20. doi: 10.1038/s41422-019-0236-6. Epub 
            2019 Oct 8.
  PUBMED    31595041
REFERENCE   2  (bases 1 to 5949)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41422-019-0236-6"; journalName: "Cell Res"; date: 
            "2020-01"; volume: "30"; issue: "1"; pages: "5-20"
COMMENT     Sequence Label: ER_K0
FEATURES             Location/Qualifiers
     source          1..5949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..338
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        339..542
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     493..513
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(704..706,707..1417)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(746..767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1007..1026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1424..1489
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1871..2578
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2013..2031)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2281..2300
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2711..2832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2748..2767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2802..2821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2839..3294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2976..2997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3188..3207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3321..3425
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3427..3784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3427..3618
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3450..3470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3635..3770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3819..4613
                     /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             3819..4613
                     /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(3873..3892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4483..4502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4801..4820)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4845..4892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5221..5809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5710..5729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
       61 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      121 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      181 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      241 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      301 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      361 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      421 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      481 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      541 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg taccgtgcac
      601 gctgctcctg ctgttggcgg ccgccctggc tccgactcag acccgcgccg gctctcactc
      661 ccaagatctt ccaggaaacg ataacagtac agcaggaggt agtgtgagca agggcgagga
      721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      781 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc
     1381 cgccgggatc actctcggca tggacgagct gtacaagtcc ggagactaca aagaccatga
     1441 cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg ccggatctcc
     1501 tggtccgacc cccagtggca ctaacgtggg atcctcactc tctgacgagg acttcaaggc
     1561 tgtttttggc atgacccgct ctgcctttgc caacttgccc ttgtggaaac agcagaacct
     1621 caagaaggag aaaggactct tcggatcagg gcgctctccc agcaaagcag tggccacccg
     1681 ggcggcggga tccactgtcc ggcagaggaa aaatgccagc tgtgggacaa ggagtgcagg
     1741 ccgcacaacc tcggcaggca ccggggggat gtggcgattc tacacagaag attcacctgg
     1801 gctcaaagtt ggccctaggt cgaccgccat ggcgaccccc gggggatccg gtaccggaag
     1861 cggactcgag gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     1921 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1981 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2041 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2101 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2161 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2221 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2281 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2341 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2401 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2461 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2521 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaac
     2581 tggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     2641 acttgcttta aaaaacctcc cacacctccc ccggaacctg aaacataaaa tgaatgcaat
     2701 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2761 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2821 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2881 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2941 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3001 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3061 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3121 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3181 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3241 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3301 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3361 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3421 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3781 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4321 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4801 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4861 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4921 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4981 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5041 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5881 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     5941 caattacgg
//
