LOCUS       sequence-269960-        7120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The Reverse Complement of TMED3 3'UTR 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-Reverse-Complement
FEATURES             Location/Qualifiers
     source          1..7120
                     /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    1899..1947
                     /note="synthetic polyadenylation signal"
     promoter        1956..2707
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2777..2909
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2954..2972
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2981..2990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2987..4639
                     /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             3946..3954
                     /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    4664..4785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4675..4694)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4729..4748
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5023..5091,5092..5883)
                     /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(5241..5260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6054..6642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6543..6562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6771..7120,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6771..6962
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6794..6814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6979..7114
                     /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 gagtttgtga aggtgaaggg cctccacttc
     1141 agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg
     1201 ctgaagaacg agcagtaatt ctaggcgatc gctcgagtct ccaggacgct cacacttagt
     1261 ttttattagc cacagtttcc cacagttttc tacctcctag gaaatacaca gctcaccaag
     1321 ggcacccagt caccattctg tcctgcttgc atggctgaca ctgttgctca ccgagggtga
     1381 gcagatctgc aaagtcaccc agggcctggt ttcctcaggt acagagaacc ccaaagaaag
     1441 aagagccaga acttagagcc cctttcttct ccatatggga taggacaccc aagacaaatg
     1501 acccatgcat catgaaacag aggcagggcc ctaagctgcc caagaggcct gggcacttgg
     1561 agttcctgcc aacagccagg ccactgaacc attgcctgtc caccctccca cagtggctaa
     1621 tccctggcct agttgtaaat gtcttttcag cgctgcacac ttaatgcagt cggatgcctt
     1681 ctgattactg gtctctgaga ccttcagctg ctgaggtgca agaatcaagc atgggcatga
     1741 aaggcaaagg gctaccacag cagcatcgtt ctgcctccac cctgggctac gcgacccagc
     1801 ccagcaggct gtgacctagg agagctgctc cagcctgggg catgaggggc cctagagcag
     1861 gatgccgggg gtttaaacct agagcggccg ctggccgcaa taaaatatct ttattttcat
     1921 tacatctgtg tgttggtttt ttgtgtgagg atctaaatga gtcttcggac ctcgcggggg
     1981 ccgcttaagc ggtggttagg gtttgtctga cgcgggggga gggggaagga acgaaacact
     2041 ctcattcgga ggcggctcgg ggtttggtct tggtggccac gggcacgcag aagagcgccg
     2101 cgatcctctt aagcaccccc ccgccctccg tggaggcggg ggtttggtcg gcgggtggta
     2161 actggcgggc cgctgactcg ggcgggtcgc gcgccccaga gtgtgacctt ttcggtctgc
     2221 tcgcagaccc ccgggcggcg ccgccgcggc ggcgacgggc tcgctgggtc ctaggctcca
     2281 tggggaccgt atacgtggac aggctctgga gcatccgcac gactgcggtg atattaccgg
     2341 agaccttctg cgggacgagc cgggtcacgc ggctgacgcg gagcgtccgt tgggcgacaa
     2401 acaccaggac ggggcacagg tacactatct tgtcacccgg aggcgcgagg gactgcagga
     2461 gcttcaggga gtggcgcagc tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg
     2521 tgtccccgga agaaatatat ttgcatgtct ttagttctat gatgacacaa accccgccca
     2581 gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt
     2641 ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc
     2701 cgcttaaaag cttgattctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
     2761 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     2821 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     2881 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     2941 aggctagagt acttaatacg actcactata ggctagcaaa gccaccatgg ccgatgctaa
     3001 gaacattaag aagggccctg ctcccttcta ccctctggag gatggcaccg ctggcgagca
     3061 gctgcacaag gccatgaaga ggtatgccct ggtgcctggc accattgcct tcaccgatgc
     3121 ccacattgag gtggacatca cctatgccga gtacttcgag atgtctgtgc gcctggccga
     3181 ggccatgaag aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc
     3241 tctgcagttc ttcatgccag tgctgggcgc cctgttcatc ggagtggccg tggcccctgc
     3301 taacgacatt tacaacgagc gcgagctgct gaacagcatg ggcatttctc agcctaccgt
     3361 ggtgttcgtg tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga agctgcctat
     3421 catccagaag atcatcatca tggactctaa gaccgactac cagggcttcc agagcatgta
     3481 cacattcgtg acatctcatc tgcctcctgg cttcaacgag tacgacttcg tgccagagtc
     3541 tttcgacagg gacaaaacca ttgccctgat catgaacagc tctgggtcta ccggcctgcc
     3601 taagggcgtg gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc
     3661 tattttcggc aaccagatca tccccgacac cgctattctg agcgtggtgc cattccacca
     3721 cggcttcggc atgttcacca ccctgggcta cctgatttgc ggctttcggg tggtgctgat
     3781 gtaccgcttc gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc
     3841 cctgctggtg ccaaccctgt tcagcttctt cgctaagagc accctgatcg acaagtacga
     3901 cctgtctaac ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga
     3961 agccgtggcc aagcgctttc atctgccagg catccgccag ggctacggcc tgaccgagac
     4021 aaccagcgcc attctgatta ccccagaggg cgacgacaag cctggcgccg tgggcaaggt
     4081 ggtgccattc ttcgaggcca aggtggtgga cctggacacc ggcaagaccc tgggagtgaa
     4141 ccagcgcggc gagctgtgtg tgcgcggccc tatgattatg tccggctacg tgaataaccc
     4201 tgaggccaca aacgccctga tcgacaagga cggctggctg cactctggcg acattgccta
     4261 ctgggacgag gacgagcact tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa
     4321 gggctaccag gtggccccag ccgagctgga gtctatcctg ctgcagcacc ctaacatttt
     4381 cgacgccgga gtggccggcc tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt
     4441 cgtgctggaa cacggcaaga ccatgaccga gaaggagatc gtggactatg tggccagcca
     4501 ggtgacaacc gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg
     4561 cctgaccggc aagctggacg cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg
     4621 cggcaagatc gccgtgtaat gcggccgctt cgagcagaca tgataagata cattgatgag
     4681 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4741 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4801 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     4861 ctctacaaat gtggtaaaat cgataaggat ccaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5701 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatctgcgca
     6721 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     6781 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     7021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     7081 ttccagaagt agtgaggagg cttttttgga ggcctaggct
//
