LOCUS       sequence-419246-        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual-luciferase reporter plasmid backbone for insertion of your TRE 
            of interest driving minTK-luc2P expression in addition to 
            constitutive SV-40-driven Renilla luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  AUTHORS   Zahm AM, Owens WS, Himes SR, Fallon BS, Rondem KE, Gormick AN, Bloom
            JS, Kosuri S, Chan H, English JG
  TITLE     A massively parallel reporter assay library to screen short 
            synthetic promoters in mammalian cells.
  JOURNAL   Nat Commun. 2024 Nov 28;15(1):10353. doi: 
            10.1038/s41467-024-54502-9.
  PUBMED    39609378
REFERENCE   2  (bases 1 to 5879)
  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"; journalName: "Nat 
            Commun."; date: "2024-11-28"; volume: "15(1):10353. doi"; pages: " 
            10.1038/s41467-024-54502-9"
COMMENT     Sequence Label: pGL4.R_TRE_minTK
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    969..1017
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1173..1190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1344..1932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1424..1443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2132..2923,2924..2992))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3097..3145
                     /note="synthetic polyadenylation signal"
     misc_feature    3159..3250
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3199..3218
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3293..3355
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     regulatory      3356..3365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3362..5011
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             5015..5134
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    5186..5307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5197..5216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5251..5270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5501..5858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5501..5692
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5524..5544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5709..5844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5771..5790
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 agcgccacca tgaccagcaa ggtgtacgac cccgagcaga ggaagaggat gatcaccggc
       61 ccccagtggt gggccaggtg caagcagatg aacgtgctgg acagcttcat caactactac
      121 gacagcgaga agcacgccga gaacgccgtg atcttcctgc acggcaacgc cgctagcagc
      181 tacctgtgga ggcacgtggt gccccacatc gagcccgtgg ccaggtgcat catccccgat
      241 ctgatcggca tgggcaagag cggcaagagc ggcaacggca gctacaggct gctggaccac
      301 tacaagtacc tgaccgcctg gttcgagctc ctgaacctgc ccaagaagat catcttcgtg
      361 ggccacgact ggggcgcctg cctggccttc cactacagct acgagcacca ggacaagatc
      421 aaggccatcg tgcacgccga gagcgtggtg gacgtgatcg agagctggga cgagtggcca
      481 gacatcgagg aggacatcgc cctgatcaag agcgaggagg gcgagaagat ggtgctggag
      541 aacaacttct tcgtggagac catgctgccc agcaagatca tgagaaagct ggagcccgag
      601 gagttcgccg cctacctgga gcccttcaag gagaagggcg aggtgagaag acccaccctg
      661 agctggccca gagagatccc cctggtgaag ggcggcaagc ccgacgtggt gcagatcgtg
      721 agaaactaca acgcctacct gagagccagc gacgacctgc ccaagatgtt catcgagagc
      781 gaccccggct tcttcagcaa cgccatcgtg gagggcgcca agaagttccc caacaccgag
      841 ttcgtgaagg tgaagggcct gcacttcagc caggaggacg cccccgacga gatgggcaag
      901 tacatcaaga gcttcgtgga gagagtgctg aagaacgagc agtaaactct agaaccggtc
      961 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc
     1021 gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     1081 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     1141 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1201 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1741 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2041 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2101 agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga
     2161 tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca
     2221 ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc
     2281 gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa
     2341 gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag
     2401 taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg
     2461 tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg
     2521 tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg
     2581 tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc
     2641 ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt
     2701 tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca
     2761 ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga
     2821 aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca
     2881 gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc
     2941 aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc
     3001 tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa
     3061 gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta
     3121 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac
     3181 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     3241 acatttctct ggcctaactg gccggtacgg cgcgccaact cgagaaggta ccttcgcata
     3301 ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaagccac
     3361 catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg
     3421 gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat
     3481 cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag
     3541 cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt
     3601 gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt
     3661 ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat
     3721 cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca
     3781 aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg
     3841 cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga
     3901 cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg
     3961 cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag
     4021 tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt
     4081 ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt
     4141 tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta
     4201 taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct
     4261 catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag
     4321 caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta
     4381 cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg
     4441 cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa
     4501 gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg
     4561 ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag
     4621 cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag
     4681 cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca
     4741 acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct
     4801 gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga
     4861 ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga
     4921 cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat
     4981 taaggccaag aagggcggca agatcgccgt gaattctcac ggcttccctc ccgaggtgga
     5041 ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag gagagcggca tggatagaca
     5101 ccctgctgct tgcgccagcg ccaggatcaa cgtctaaggc cgcgactcta gagtcggggc
     5161 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     5221 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5281 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5341 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     5401 atcgataagg atccgtttgc gtattgggcg ctcttccgct gatctgcgca gcaccatggc
     5461 ctgaaataac ctctgaaaga ggaacttggt tagctacctt ctgaggcgga aagaaccagc
     5521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5761 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct tctgacact
//
