LOCUS       sequence-449375-        5478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cDELE1 (corresponding to mitochondrially imported, cleaved DELE1, 
            residues 142-515) fused to sumo used for in-vitro translation by 
            wheat germ extract in in-vitro ubiquitylation assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5478)
  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 5478)
  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)-sumo
FEATURES             Location/Qualifiers
     source          1..5478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..55)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(39..55)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(52..74)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    63..79
                     /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)."
     promoter        complement(join(87..93,94..111,112..117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    132..153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(270..287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(441..1029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(521..540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1200..1991,1992..2060))
                     /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     1823..1842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2061..2165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2191..2646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2328..2349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2540..2559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2717..3701
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3736..3754
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3736..3753
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4911..5198
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLTFLYDGIEIQADQTPEDLDMEDNDIIEAHREQIGG"
     polyA_signal    5231..5365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5255..5274)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5309..5328
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(join(5478,1..20))
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(join(5478,1..18))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
       61 aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc
      121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2161 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     2221 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2281 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2341 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2401 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2461 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2521 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2581 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2641 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2701 cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt
     2761 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
     2821 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
     2881 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
     2941 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
     3001 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
     3061 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
     3121 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
     3181 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
     3241 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
     3301 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
     3361 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
     3421 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
     3481 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
     3541 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     3601 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     3661 cctaatggga ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac
     3721 gtcggagcaa gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag
     3781 gatccatgca catcctcccc agccccgatg gcccagctcc caggcacact ggcctcaggg
     3841 aacccaggct tggccaggaa gaagcctcag ctcagccccg gaacttctca cacaactctt
     3901 tgagaggagc tcgtcctcag gacccctctg aggaaggtcc cggtgatttt ggcttcctgc
     3961 atgccagtag tagcatcgag tccgaggcaa aaccagccca gcctcagccc actggtgaaa
     4021 aggaacaaga taaatcaaaa actctttccc ttgaggaggc tgtgacttcc attcagcagc
     4081 tcttccagct cagtgtttcc atcactttca acttcctggg aacagagaac atgaagagtg
     4141 gcgaccacac ggcagccttt tcttacttcc agaaagctgc agcccgcggc tacagcaaag
     4201 cgcagtacaa tgcgggcttg tgtcatgagc atggcagagg cacccccagg gacattagca
     4261 aggcggtcct ttattatcag ttggctgcca gccagggcca cagcctggct cagtaccgct
     4321 atgccaggtg cctactacga gacccagcct cttcgtggaa ccctgagcgg cagagggcag
     4381 tgtccttgct gaagcaggct gcagactcag gcttgagaga ggcccaagct ttcctcgggg
     4441 tgcttttcac caaggagccc tacctggatg agcagagagc tgtgaaatat ctttggcttg
     4501 cagccaacaa tggggactca cagagcaggt accaccttgg aatttgctat gagaaaggcc
     4561 ttggtgtgca gaggaatctg ggagaggcct tgagatgtta ccagcagtca gccgctctgg
     4621 gaaatgaggc cgcccaggag aggctgcgag ccctcttttc catgggggct gcagccccgg
     4681 ggcccagcga cctgacagtt acaggactga agtctttctc cagcccctcc ctctgcagct
     4741 tgaacaccct gctagcagga acctcacgcc taccacatgc ctcgagcaca ggcaaccttg
     4801 gcctcctctg cagaagtggg catctcggag ccagcctgga agcctccagc agggctattc
     4861 ccccacaccc ctacccactg gaaaggagtg ttgtaagact aggttttggc gactcagaag
     4921 tcaatcaaga agctaagcca gaggtcaagc cagaagtcaa gcctgagact cacatcaatt
     4981 taaaggtgtc cgatggatct tcagagatct tcttcaagat caaaaagacc actcctttaa
     5041 gaaggctgat ggaagcgttc gctaaaagac agggtaagga aatggactcc ttaacattct
     5101 tgtacgacgg tattgaaatt caagctgatc agacccctga agatttggac atggaggata
     5161 acgatattat tgaggctcac cgcgaacaga ttggaggtta gtctagaact atagtgagtc
     5221 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5281 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5341 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     5401 tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg
     5461 gtacccagct tttgttcc
//
