LOCUS       sequence-448403-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cDELE1 (corresponding to mitochondrially imported, cleaved DELE1, 
            residues 142-515) stability reporter construct for transient 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  AUTHORS   Haakonsen DL, Heider M, Ingersoll AJ, Vodehnal K, Witus SR, Uenaka 
            T, Wernig M, Rape M
  TITLE     Stress response silencing by an E3 ligase mutated in 
            neurodegeneration.
  JOURNAL   Nature. 2024 Feb;626(8000):874-880. doi: 10.1038/s41586-023-06985-7.
            Epub 2024 Jan 31.
  PUBMED    38297121
REFERENCE   2  (bases 1 to 7227)
  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/s41586-023-06985-7"; journalName: "Nature"; date: 
            "2024-02"; volume: "626"; issue: "8000"; pages: "874-880"
COMMENT     Sequence Label: pcs2-cDELE1(142-515)-GFP-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..7227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     248..265
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(440..1159)
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     promoter        complement(2337..2355)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2338..2355)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2390..3374)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      3445..3900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3532..3551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3742..3763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3926..4030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4031..4099,4100..4891)
                     /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(4249..4268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5062..5650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5551..5570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5804..5821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5938..5959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5974..5979,5980..5997,5998..6004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6012..6028
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6017..6039
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6036..6052
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6036..6052
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6071..6091
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6073..6091
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    6204..6338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6241..6260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6295..6314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6368..7078)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     6915..6933
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    join(7087..7227,1..432)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
ORIGIN
        1 tccttcagcc ccttgttgaa tacgcttgag gagagccatt tgactctttc cacaactatc
       61 caactcacaa cgtggcactg gggttgtgcc gcctttgcag gtgtatctta tacacgtggc
      121 ttttggccgc agaggcacct gtcgccaggt ggggggttcc gctgcctgca aagggtcgct
      181 acagacgttg tttgtcttca agaagcttcc agaggaactg cttccttcac gacattcaac
      241 agaccttgca ttcctttggc gagaggggaa agacccctag gaatgctcgt caagaagaca
      301 gggccaggtt tccgggccct cacattgcca aaagacggca atatggtgga aaataacata
      361 tagacaaacg cacaccggcc ttattccaag cggcttcggc cagtaacgtt aggggggggg
      421 gagggagagg ggctctagat cacttgtaca gctcatccat gccgtgggtg atggcggcgg
      481 cggtcacgaa gccgaagtag atcatgtgat cgcgcttctc gttggggtcc ttggacaggg
      541 cgctctgggt ggacaggtag tggttatcgg gcagcagcac agggccatcg ccgatggggg
      601 tattctgctg gtagtggtcg gccagctgca cgctgccatc ctcgatgttg tggcggatct
      661 tgaagttcac cttgatgcca ttcttggcct tgtcggtcat gatgtacaca ttgtgggcgt
      721 tgtagttgta ctccatctta ttgcccagga tgttgccatc ctccttgaaa tcggtgccgg
      781 tcagctcgat gcgattcacc agggtatcgc cctcgaactt cacctcggcg cgcgacttgt
      841 agttgccgtc atcctcgaag aagatggtgc gctcctggat gtagccctca ggcatggcgc
      901 tcttgaagaa gtcgtgctgc ttcatgtgat cggggtagcg tgagaagcac tgcacgccgt
      961 agctcagggt ggtcaccagg gtgggccagg gcacaggcag cttgccggtg gtgcagatga
     1021 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccgctc acgctgaact
     1081 tgtggccatt cacatcgcca ttcagctcga tcaggatggg cacgatgccg gtgaacagct
     1141 cggcgccctt gctcaccatg ctgcctccgc ctccgccaaa acctagtctt acaacactcc
     1201 tttccagtgg gtaggggtgt gggggaatag ccctgctgga ggcttccagg ctggctccga
     1261 gatgcccact tctgcagagg aggccaaggt tgcctgtgct cgaggcatgt ggtaggcgtg
     1321 aggttcctgc tagcagggtg ttcaagctgc agagggaggg gctggagaaa gacttcagtc
     1381 ctgtaactgt caggtcgctg ggccccgggg ctgcagcccc catggaaaag agggctcgca
     1441 gcctctcctg ggcggcctca tttcccagag cggctgactg ctggtaacat ctcaaggcct
     1501 ctcccagatt cctctgcaca ccaaggcctt tctcatagca aattccaagg tggtacctgc
     1561 tctgtgagtc cccattgttg gctgcaagcc aaagatattt cacagctctc tgctcatcca
     1621 ggtagggctc cttggtgaaa agcaccccga ggaaagcttg ggcctctctc aagcctgagt
     1681 ctgcagcctg cttcagcaag gacactgccc tctgccgctc agggttccac gaagaggctg
     1741 ggtctcgtag taggcacctg gcatagcggt actgagccag gctgtggccc tggctggcag
     1801 ccaactgata ataaaggacc gccttgctaa tgtccctggg ggtgcctctg ccatgctcat
     1861 gacacaagcc cgcattgtac tgcgctttgc tgtagccgcg ggctgcagct ttctggaagt
     1921 aagaaaaggc tgccgtgtgg tcgccactct tcatgttctc tgttcccagg aagttgaaag
     1981 tgatggaaac actgagctgg aagagctgct gaatggaagt cacagcctcc tcaagggaaa
     2041 gagtttttga tttatcttgt tccttttcac cagtgggctg aggctgggct ggttttgcct
     2101 cggactcgat gctactactg gcatgcagga agccaaaatc accgggacct tcctcagagg
     2161 ggtcctgagg acgagctcct ctcaaagagt tgtgtgagaa gttccggggc tgagctgagg
     2221 cttcttcctg gccaagcctg ggttccctga ggccagtgtg cctgggagct gggccatcgg
     2281 ggctggggag gatgtgcatg gtggcgggat ccgcaaaaag aacaagtagc ttgtattcta
     2341 tagtgtcacc taaatcaagc ttgctccgac gtccccaggc agaatggcgg ttccctaaac
     2401 gagcattgct tatatagacc tcccattagg cacgcctacc gcccatttac gtcaatggaa
     2461 cgcccatttg cgtcattgcc cctccccatt gacgtcaatg gggatgtact tggcagccat
     2521 cgcgggccat ttaccgccat tgacgtcaat gggagtactg ccaatgtacc ctggcgtact
     2581 tccaatagta atgtacttgc caagttacta ttaatagata ttgatgtact gccaagtggg
     2641 ccatttaccg tcattgacgt caataggggg cgtgagaacg gatatgaatg ggcaatgagc
     2701 catcccattg acgtcaatgg tgggtggtcc tattgacgtc aatgggcatt gagccaggcg
     2761 ggccatttac cgtaattgac gtcaatgggg gaggcgccat atacgtcaat aggaccgccc
     2821 atatgacgtc aataggaaag accatatata gagaccattg acgtcaatgg gggagtggct
     2881 atgggcggta ttaggaagcc ccatatatgg tatatgggac cgcccattgg gaggggctat
     2941 ctacgtcaat aggaaaaccc atatatggaa tactatatgg catagggcca atacatagta
     3001 ttgaacctgg ccaatagcca tattggcata gggccatatt ggatattgcc tatatattga
     3061 tcctggcata tagccaatat ggccgccatt attggcacca tgccaattca atatggcgga
     3121 cttggcacag ggccaagacc cctcctcaaa cttggcaccg tgccaagtag acccctcccc
     3181 agttggcaca gtgccaggtc cgccatattg aattggcatg gtgccaagtc cgccatattg
     3241 aattggctgg ggtccagatc caccatattg aattggcacg agtccatata cgccatattg
     3301 aattggcaca ggtccagatc caccatattg aattggcatg agtccatata cgccatattg
     3361 aattggcggt cgactggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3421 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3481 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3541 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3601 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3661 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3721 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3781 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3841 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3901 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3961 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4021 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4081 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4141 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4201 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4261 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4321 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4381 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4441 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4501 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4561 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4621 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4681 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4741 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4801 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4861 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5761 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5821 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5881 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5941 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgctcc cggctcgtat
     6001 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6061 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctgggtaccg ggcccaatgc
     6121 attggcgccg cggccgcgaa ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6181 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6301 tgtccaaact catcaatgta tcttatcatg tctggatcta cgtaatacga ctcactatag
     6361 ttctagatca cttgtacagc tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt
     6421 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga
     6481 cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcag
     6541 cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc
     6601 cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggttttc ttctgcatta
     6661 cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt
     6721 cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc
     6781 gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt
     6841 gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg
     6901 gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     6961 cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct
     7021 tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccattc
     7081 cggaggtatt atcgtgtttt tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga
     7141 cgttttttta acctcgacta aacacatgta aagcatgtgc accgaggccc cagatcagat
     7201 cccatacaat ggggtacctt ctgggca
//
