LOCUS       sequence-450772-        6151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sUPRa is a dual promoter and two-color construct, which is designed 
            to report the global unfolded protein response (UPR) and affords 
            unbiased cell detection irrespective of UPR levels.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6151)
  AUTHORS   Chakrabarty A, Newey SE, Promi MM, Agbetiameh BK, Munro D, Brodersen
            PJN, Gothard G, Mahfooz K, Mengual JP, Vyazovskiy VV, Akerman CJ
  TITLE     sUPRa is a dual-color reporter for unbiased quantification of the 
            unfolded protein response with cellular resolution.
  JOURNAL   Sci Rep. 2024 Jul 1;14(1):14990. doi: 10.1038/s41598-024-65611-2.
  PUBMED    38951511
REFERENCE   2  (bases 1 to 6151)
  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"; journalName: "Sci Rep."; 
            date: "2024-07-1"; volume: "14(1):14990. doi"; pages: " 
            10.1038/s41598-024-65611-2"
COMMENT     Sequence Label: sUPRa
FEATURES             Location/Qualifiers
     source          1..6151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      338..347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             344..1051
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1186..1662
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1385..1404)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     promoter        1718..1929
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1942..2210
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      2238..2247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2244..2939
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2377..2395)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2645..2664
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3012..3032)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3414..3554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3414..3543
                     /label=AAV2 ITR
     rep_origin      3629..4084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3716..3735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3926..3947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4101..4120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4220..4242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4280..4298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4366..4470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4471..4539,4540..5331)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4689..4708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5502..6090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5991..6010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttagcaa tgacgtgagt tgcggaggag gccgcttcga
      181 atcggcagca gccagcttgg tggcatggac caatcagcgg cctccaacga gtagcgactt
      241 caccaatcgg aggcctccac gacggggctg tggggagggt atataaggcg agtcggcgac
      301 ggcgcgctcg atactggccg agacaacact gggtaccgcc accatggtga gcaagggcga
      361 ggaggataac atggcctctc tcccagcgac acatgagtta cacatctttg gctccatcaa
      421 cggtgtggac tttgacatgg tgggtcaggg caccggcaat ccaaatgatg gttatgagga
      481 gttaaacctg aagtccacca agggtgacct ccagttctcc ccctggattc tggtccctca
      541 tatcgggtat ggcttccatc agtacctgcc ctaccctgac gggatgtcgc ctttccaggc
      601 cgccatggta gatggctccg gataccaagt ccatcgcaca atgcagtttg aagatggtgc
      661 ctcccttact gttaactacc gctacaccta cgagggaagc cacatcaaag gagaggccca
      721 ggtgaagggg actggtttcc ctgctgacgg tcctgtgatg accaactcgc tgaccgctgc
      781 ggactggtgc aggtcgaaga agacttaccc caacgacaaa accatcatca gtacctttaa
      841 gtggagttac accactggaa atggcaagcg ctaccggagc actgcgcgga ccacctacac
      901 ctttgccaag ccaatggcgg ctaactatct gaagaaccag ccgatgtacg tgttccgtaa
      961 gacggagctc aagcactcca agaccgagct caacttcaag gagtggcaaa aggcctttac
     1021 cgatgtgatg ggcatggacg agctgtacaa gcatggcttc ccgccggagg tggaggagca
     1081 ggatgatggc acgctgccca tgtcttgtgc ccaggagagc gggatggacc gtcaccctgc
     1141 agcctgtgct tctgctagga tcaatgtgta gctcgagaga tctacgggtg gcatccctgt
     1201 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     1261 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     1321 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     1381 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     1441 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     1501 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     1561 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     1621 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     1681 acgtgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     1741 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc
     1801 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     1861 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     1921 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc
     1981 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     2041 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     2101 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     2161 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga
     2221 ccggcgccta cgtcgaggcc accatggtga gcaagggcga ggcagtgatc aaggagttca
     2281 tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     2341 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     2401 gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct
     2461 tcatcaagca ccccgccgac atccccgact actataagca gtccttcccc gagggcttca
     2521 agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc caggacacct
     2581 ccctggagga cggcaccctg atctacaagg tgaagctccg cggcaccaac ttccctcctg
     2641 acggccccgt aatgcagaag aagacaatgg gctgggaagc gtccaccgag cggttgtacc
     2701 ccgaggacgg cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc
     2761 gctacctggc ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg
     2821 cctacaacgt cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg
     2881 aacagtacga acgctccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     2941 aagaattccg ctcgagataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3001 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3061 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     3121 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3181 ttgggcactg acaattccgt ggtgtttatt tgtgaaattt gtgatgctat tgctttattt
     3241 gtaaccatct agctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3301 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3361 ggagatgtgg gaggtttttt aaagcggccg ccacgtgcgg accgagcggc cgcaggaacc
     3421 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3481 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3541 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3601 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     3721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3841 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3961 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4021 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4081 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4141 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4201 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4261 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4321 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4381 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4441 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4501 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4561 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4621 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4681 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4741 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4801 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4861 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4921 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4981 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5041 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5101 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5161 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5221 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5281 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5341 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5401 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5461 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5521 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5581 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5641 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5701 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5761 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5821 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5881 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5941 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6001 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6061 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6121 ctggcctttt gctggccttt tgctcacatg t
//
