LOCUS       sequence-269575-        7034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SDHAF3 3'UTR (NM_020186.2) inserted into the 3'UTR of Renilla 
            luciferase in the psiCHECK3 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  AUTHORS   Fischer JW, Busa VF, Shao Y, Leung AKL
  TITLE     Structure-Mediated RNA Decay by UPF1 and G3BP1.
  JOURNAL   Mol Cell. 2020 Apr 2;78(1):70-84.e6. doi: 
            10.1016/j.molcel.2020.01.021. Epub 2020 Feb 3.
  PUBMED    32017897
REFERENCE   2  (bases 1 to 7034)
  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: 
            "10.1016/j.molcel.2020.01.021"; journalName: "Mol Cell"; date: 
            "2020-04-2- 2"; volume: "78"; issue: "1"; pages: "70-84.e6"
COMMENT     Sequence Label: psiCHECK3-SDHAF3
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          78..210
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        255..273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      277..286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             283..1218
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1813..1861
                     /note="synthetic polyadenylation signal"
     promoter        1870..2621
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2691..2823
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2868..2886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2895..2904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2901..4553
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3860..3868
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4578..4699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4589..4608)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4643..4662
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4832..4936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4937..5005,5006..5797)
                     /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(5155..5174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5968..6556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6457..6476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6685..7034,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6685..6876
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6708..6728)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6893..7028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt
       61 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
      121 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
      181 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt
      241 aaggctagag tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac
      301 gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa
      361 atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc
      421 gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac
      481 atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag
      541 agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag
      601 ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc
      661 tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc
      721 gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc
      781 aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc
      841 ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc
      901 aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt
      961 aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc
     1021 agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt
     1081 gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc
     1141 agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg
     1201 ctgaagaacg agcagtaatt ctaggcgatc gctcgagtct atacaacaaa gcttaataag
     1261 acatgcaaaa atttagaacc cctactttaa ctgtcattgg tttttgaaat atatttaagc
     1321 tttgaaaaca cctgttatta atgaaatact cttttatttt ggatattatg attgcagtat
     1381 atggatcaag atcactagtg acaattgaaa aaaactattg gaataatagc acttgtataa
     1441 aattcagttt tggaactaaa cagcaaattt ctagaatttt gctgaaaatg ttttaaaatg
     1501 ctattctcat ccagccatat tagtcttctg gcttttcttt agcttcatca aataagcatg
     1561 ttgtgataat gatagatgta caattccaac aaggttatta ttttttaaat acattgtcat
     1621 tctgaacatt ttatcacttc tagtttaata atacatacat gatttttctt ctgaatgtct
     1681 cttctccctg catcactgtt cattcacaat gaaaggttag gaagaagctt taaaattcac
     1741 tattttacta tcaatcattt gtataataaa ctatacaaag tatagtttaa acctagagcg
     1801 gccgctggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     1861 gaggatctaa atgagtcttc ggacctcgcg ggggccgctt aagcggtggt tagggtttgt
     1921 ctgacgcggg gggaggggga aggaacgaaa cactctcatt cggaggcggc tcggggtttg
     1981 gtcttggtgg ccacgggcac gcagaagagc gccgcgatcc tcttaagcac ccccccgccc
     2041 tccgtggagg cgggggtttg gtcggcgggt ggtaactggc gggccgctga ctcgggcggg
     2101 tcgcgcgccc cagagtgtga ccttttcggt ctgctcgcag acccccgggc ggcgccgccg
     2161 cggcggcgac gggctcgctg ggtcctaggc tccatgggga ccgtatacgt ggacaggctc
     2221 tggagcatcc gcacgactgc ggtgatatta ccggagacct tctgcgggac gagccgggtc
     2281 acgcggctga cgcggagcgt ccgttgggcg acaaacacca ggacggggca caggtacact
     2341 atcttgtcac ccggaggcgc gagggactgc aggagcttca gggagtggcg cagctgcttc
     2401 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     2461 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     2521 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     2581 gtggcctcga acaccgagcg accctgcagc gacccgctta aaagcttgat tcttctgaca
     2641 caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
     2701 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     2761 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     2821 caggtgtcca ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac
     2881 tataggctag caaagccacc atggccgatg ctaagaacat taagaagggc cctgctccct
     2941 tctaccctct ggaggatggc accgctggcg agcagctgca caaggccatg aagaggtatg
     3001 ccctggtgcc tggcaccatt gccttcaccg atgcccacat tgaggtggac atcacctatg
     3061 ccgagtactt cgagatgtct gtgcgcctgg ccgaggccat gaagaggtac ggcctgaaca
     3121 ccaaccaccg catcgtggtg tgctctgaga actctctgca gttcttcatg ccagtgctgg
     3181 gcgccctgtt catcggagtg gccgtggccc ctgctaacga catttacaac gagcgcgagc
     3241 tgctgaacag catgggcatt tctcagccta ccgtggtgtt cgtgtctaag aagggcctgc
     3301 agaagatcct gaacgtgcag aagaagctgc ctatcatcca gaagatcatc atcatggact
     3361 ctaagaccga ctaccagggc ttccagagca tgtacacatt cgtgacatct catctgcctc
     3421 ctggcttcaa cgagtacgac ttcgtgccag agtctttcga cagggacaaa accattgccc
     3481 tgatcatgaa cagctctggg tctaccggcc tgcctaaggg cgtggccctg cctcatcgca
     3541 ccgcctgtgt gcgcttctct cacgcccgcg accctatttt cggcaaccag atcatccccg
     3601 acaccgctat tctgagcgtg gtgccattcc accacggctt cggcatgttc accaccctgg
     3661 gctacctgat ttgcggcttt cgggtggtgc tgatgtaccg cttcgaggag gagctgttcc
     3721 tgcgcagcct gcaagactac aaaattcagt ctgccctgct ggtgccaacc ctgttcagct
     3781 tcttcgctaa gagcaccctg atcgacaagt acgacctgtc taacctgcac gagattgcct
     3841 ctggcggcgc cccactgtct aaggaggtgg gcgaagccgt ggccaagcgc tttcatctgc
     3901 caggcatccg ccagggctac ggcctgaccg agacaaccag cgccattctg attaccccag
     3961 agggcgacga caagcctggc gccgtgggca aggtggtgcc attcttcgag gccaaggtgg
     4021 tggacctgga caccggcaag accctgggag tgaaccagcg cggcgagctg tgtgtgcgcg
     4081 gccctatgat tatgtccggc tacgtgaata accctgaggc cacaaacgcc ctgatcgaca
     4141 aggacggctg gctgcactct ggcgacattg cctactggga cgaggacgag cacttcttca
     4201 tcgtggaccg cctgaagtct ctgatcaagt acaagggcta ccaggtggcc ccagccgagc
     4261 tggagtctat cctgctgcag caccctaaca ttttcgacgc cggagtggcc ggcctgcccg
     4321 acgacgatgc cggcgagctg cctgccgccg tcgtcgtgct ggaacacggc aagaccatga
     4381 ccgagaagga gatcgtggac tatgtggcca gccaggtgac aaccgccaag aagctgcgcg
     4441 gcggagtggt gttcgtggac gaggtgccca agggcctgac cggcaagctg gacgcccgca
     4501 agatccgcga gatcctgatc aaggctaaga aaggcggcaa gatcgccgtg taatgcggcc
     4561 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4621 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4681 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4741 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     4801 ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4921 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4981 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5041 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5101 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5161 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5221 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5281 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5341 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5401 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5461 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5521 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5581 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5641 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5701 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5761 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5941 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6001 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6061 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6541 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6601 gccttttgct cacatggctc gacagatctg cgcagcacca tggcctgaaa taacctctga
     6661 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     6721 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6781 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6841 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6901 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6961 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7021 tggaggccta ggct
//
