LOCUS       sequence-419254-        5875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual-luciferase reporter plasmid backbone for insertion of your TRE 
            of interest driving minCMV-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 5875)
  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 5875)
  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_minCMV
FEATURES             Location/Qualifiers
     source          1..5875
                     /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        3291..3328
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3325..3349
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3352..3361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3358..5007
                     /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             5011..5130
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    5182..5303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5193..5212)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5247..5266
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5497..5854
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5497..5688
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5520..5540)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5705..5840
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5767..5786
                     /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 gccggtacac gcgtaactcg agaaggtacc gtaggcgtgt
     3301 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcc agccaccatg
     3361 gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga agacgggacc
     3421 gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg caccatcgcc
     3481 tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga gatgagcgtt
     3541 cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat cgtggtgtgc
     3601 agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat cggtgtggct
     3661 gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat gggcatcagc
     3721 cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag
     3781 aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta ccagggcttc
     3841 caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga gtacgacttc
     3901 gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag tagtggcagt
     3961 accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg attcagtcat
     4021 gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct cagcgtggtg
     4081 ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg cggctttcgg
     4141 gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca agactataag
     4201 attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag cactctcatc
     4261 gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc gctcagcaag
     4321 gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca gggctacggc
     4381 ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa gcctggcgca
     4441 gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac cggtaagaca
     4501 ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat gagcggctac
     4561 gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct gcacagcggc
     4621 gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct gaagagcctg
     4681 atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct gctgcaacac
     4741 cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc
     4801 gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat cgtggactat
     4861 gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag
     4921 gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat tctcattaag
     4981 gccaagaagg gcggcaagat cgccgtgaat tctcacggct tccctcccga ggtggaggag
     5041 caggccgccg gcaccctgcc catgagctgc gcccaggaga gcggcatgga tagacaccct
     5101 gctgcttgcg ccagcgccag gatcaacgtc taaggccgcg actctagagt cggggcggcc
     5161 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5221 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5281 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     5341 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     5401 ataaggatcc gtttgcgtat tgggcgctct tccgctgatc tgcgcagcac catggcctga
     5461 aataacctct gaaagaggaa cttggttagc taccttctga ggcggaaaga accagctgtg
     5521 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5581 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     5641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5701 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5761 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5821 aggaggcttt tttggaggcc taggcttttg caaaaagctc gattcttctg acact
//
