LOCUS       sequence-269577-        7120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TMED3 3'UTR (NM_007364.3) 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 7120)
  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 7120)
  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-TMED3
FEATURES             Location/Qualifiers
     source          1..7120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        66..257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(89..109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      274..409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          493..625
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        670..688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             698..1633
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2314..2362
                     /note="synthetic polyadenylation signal"
     promoter        2371..3122
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3192..3324
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        3369..3387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3396..3405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3402..5054
                     /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             4361..4369
                     /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    5079..5200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5090..5109)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5144..5163
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5333..5437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5438..5506,5507..6298)
                     /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(5656..5675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6469..7057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6958..6977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgacagatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
       61 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      421 aaaaagcttg attcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga
      481 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      541 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      601 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      661 tagagtactt aatacgactc actataggct agccaccatg gcttccaagg tgtacgaccc
      721 cgagcaacgc aaacgcatga tcactgggcc tcagtggtgg gctcgctgca agcaaatgaa
      781 cgtgctggac tccttcatca actactatga ttccgagaag cacgccgaga acgccgtgat
      841 ttttctgcat ggtaacgctg cctccagcta cctgtggagg cacgtcgtgc ctcacatcga
      901 gcccgtggct agatgcatca tccctgatct gatcggaatg ggtaagtccg gcaagagcgg
      961 gaatggctca tatcgcctcc tggatcacta caagtacctc accgcttggt tcgagctgct
     1021 gaaccttcca aagaaaatca tctttgtggg ccacgactgg ggggcttgtc tggcctttca
     1081 ctactcctac gagcaccaag acaagatcaa ggccatcgtc catgctgaga gtgtcgtgga
     1141 cgtgatcgag tcctgggacg agtggcctga catcgaggag gatatcgccc tgatcaagag
     1201 cgaagagggc gagaaaatgg tgcttgagaa taacttcttc gtcgagacca tgctcccaag
     1261 caagatcatg cggaaactgg agcctgagga gttcgctgcc tacctggagc cattcaagga
     1321 gaagggcgag gttagacggc ctaccctctc ctggcctcgc gagatccctc tcgttaaggg
     1381 aggcaagccc gacgtcgtcc agattgtccg caactacaac gcctaccttc gggccagcga
     1441 cgatctgcct aagatgttca tcgagtccga ccctgggttc ttttccaacg ctattgtcga
     1501 gggagctaag aagttcccta acaccgagtt cgtgaaggtg aagggcctcc acttcagcca
     1561 ggaggacgct ccagatgaaa tgggtaagta catcaagagc ttcgtggagc gcgtgctgaa
     1621 gaacgagcag taattctagg cgatcgctcg agccccggca tcctgctcta gggcccctca
     1681 tgccccaggc tggagcagct ctcctaggtc acagcctgct gggctgggtc gcgtagccca
     1741 gggtggaggc agaacgatgc tgctgtggta gccctttgcc tttcatgccc atgcttgatt
     1801 cttgcacctc agcagctgaa ggtctcagag accagtaatc agaaggcatc cgactgcatt
     1861 aagtgtgcag cgctgaaaag acatttacaa ctaggccagg gattagccac tgtgggaggg
     1921 tggacaggca atggttcagt ggcctggctg ttggcaggaa ctccaagtgc ccaggcctct
     1981 tgggcagctt agggccctgc ctctgtttca tgatgcatgg gtcatttgtc ttgggtgtcc
     2041 tatcccatat ggagaagaaa ggggctctaa gttctggctc ttctttcttt ggggttctct
     2101 gtacctgagg aaaccaggcc ctgggtgact ttgcagatct gctcaccctc ggtgagcaac
     2161 agtgtcagcc atgcaagcag gacagaatgg tgactgggtg cccttggtga gctgtgtatt
     2221 tcctaggagg tagaaaactg tgggaaactg tggctaataa aaactaagtg tgagcgtcct
     2281 ggagagttta aacctagagc ggccgctggc cgcaataaaa tatctttatt ttcattacat
     2341 ctgtgtgttg gttttttgtg tgaggatcta aatgagtctt cggacctcgc gggggccgct
     2401 taagcggtgg ttagggtttg tctgacgcgg ggggaggggg aaggaacgaa acactctcat
     2461 tcggaggcgg ctcggggttt ggtcttggtg gccacgggca cgcagaagag cgccgcgatc
     2521 ctcttaagca cccccccgcc ctccgtggag gcgggggttt ggtcggcggg tggtaactgg
     2581 cgggccgctg actcgggcgg gtcgcgcgcc ccagagtgtg accttttcgg tctgctcgca
     2641 gacccccggg cggcgccgcc gcggcggcga cgggctcgct gggtcctagg ctccatgggg
     2701 accgtatacg tggacaggct ctggagcatc cgcacgactg cggtgatatt accggagacc
     2761 ttctgcggga cgagccgggt cacgcggctg acgcggagcg tccgttgggc gacaaacacc
     2821 aggacggggc acaggtacac tatcttgtca cccggaggcg cgagggactg caggagcttc
     2881 agggagtggc gcagctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc
     2941 ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc
     3001 ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact
     3061 tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt
     3121 aaaagcttga ttcttctgac acaacagtct cgaacttaag ctgcagaagt tggtcgtgag
     3181 gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     3241 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     3301 tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc tcttaaggct
     3361 agagtactta atacgactca ctataggcta gcaaagccac catggccgat gctaagaaca
     3421 ttaagaaggg ccctgctccc ttctaccctc tggaggatgg caccgctggc gagcagctgc
     3481 acaaggccat gaagaggtat gccctggtgc ctggcaccat tgccttcacc gatgcccaca
     3541 ttgaggtgga catcacctat gccgagtact tcgagatgtc tgtgcgcctg gccgaggcca
     3601 tgaagaggta cggcctgaac accaaccacc gcatcgtggt gtgctctgag aactctctgc
     3661 agttcttcat gccagtgctg ggcgccctgt tcatcggagt ggccgtggcc cctgctaacg
     3721 acatttacaa cgagcgcgag ctgctgaaca gcatgggcat ttctcagcct accgtggtgt
     3781 tcgtgtctaa gaagggcctg cagaagatcc tgaacgtgca gaagaagctg cctatcatcc
     3841 agaagatcat catcatggac tctaagaccg actaccaggg cttccagagc atgtacacat
     3901 tcgtgacatc tcatctgcct cctggcttca acgagtacga cttcgtgcca gagtctttcg
     3961 acagggacaa aaccattgcc ctgatcatga acagctctgg gtctaccggc ctgcctaagg
     4021 gcgtggccct gcctcatcgc accgcctgtg tgcgcttctc tcacgcccgc gaccctattt
     4081 tcggcaacca gatcatcccc gacaccgcta ttctgagcgt ggtgccattc caccacggct
     4141 tcggcatgtt caccaccctg ggctacctga tttgcggctt tcgggtggtg ctgatgtacc
     4201 gcttcgagga ggagctgttc ctgcgcagcc tgcaagacta caaaattcag tctgccctgc
     4261 tggtgccaac cctgttcagc ttcttcgcta agagcaccct gatcgacaag tacgacctgt
     4321 ctaacctgca cgagattgcc tctggcggcg ccccactgtc taaggaggtg ggcgaagccg
     4381 tggccaagcg ctttcatctg ccaggcatcc gccagggcta cggcctgacc gagacaacca
     4441 gcgccattct gattacccca gagggcgacg acaagcctgg cgccgtgggc aaggtggtgc
     4501 cattcttcga ggccaaggtg gtggacctgg acaccggcaa gaccctggga gtgaaccagc
     4561 gcggcgagct gtgtgtgcgc ggccctatga ttatgtccgg ctacgtgaat aaccctgagg
     4621 ccacaaacgc cctgatcgac aaggacggct ggctgcactc tggcgacatt gcctactggg
     4681 acgaggacga gcacttcttc atcgtggacc gcctgaagtc tctgatcaag tacaagggct
     4741 accaggtggc cccagccgag ctggagtcta tcctgctgca gcaccctaac attttcgacg
     4801 ccggagtggc cggcctgccc gacgacgatg ccggcgagct gcctgccgcc gtcgtcgtgc
     4861 tggaacacgg caagaccatg accgagaagg agatcgtgga ctatgtggcc agccaggtga
     4921 caaccgccaa gaagctgcgc ggcggagtgg tgttcgtgga cgaggtgccc aagggcctga
     4981 ccggcaagct ggacgcccgc aagatccgcg agatcctgat caaggctaag aaaggcggca
     5041 agatcgccgt gtaatgcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     5101 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5161 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5221 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5281 caaatgtggt aaaatcgata aggatccagg tggcactttt cggggaaatg tgcgcggaac
     5341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7081 acggttcctg gccttttgct ggccttttgc tcacatggct
//
