LOCUS       sequence-493245-        7112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acyclovir-regulated Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7112)
  AUTHORS   Hou Q, Oleynikov M, Mei X, Dong L, Hagen T, Jaffrey SR
  TITLE     A portable poison exon for small-molecule control of mammalian gene 
            expression.
  JOURNAL   Nat Methods. 2025 Nov 3. doi: 10.1038/s41592-025-02860-7.
  PUBMED    41184552
REFERENCE   2  (bases 1 to 7112)
  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 
            Methods. 2025 Nov 3. doi: 10.1038/s41592-025-02860-7."
COMMENT     Sequence Label: psiCHECK-2 SV40 Cyclone-RLuc
FEATURES             Location/Qualifiers
     source          1..7112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2527..2575
                     /note="synthetic polyadenylation signal"
     promoter        2584..3335
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3365..3374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3371..5023
                     /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             4330..4338
                     /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    5067..5188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5078..5097)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5132..5151
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5321..5425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5426..5494,5495..6286)
                     /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(5644..5663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6457..7045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6946..6965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggta
     1561 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     1621 agaagactct tgcgtttctg ataggcacct attggtctga tcatattttg ttgaataaaa
     1681 taagtaaaat gtcttgtgaa acaaaatgct ttttaacatc catataaagc tatctatata
     1741 tagctatcta tgtctatata gctatttttt ttaacttcct ttattttcct tacagggttt
     1801 cagacaaaat caaaaagaag gaaggtgctc acagccaagc aattaaggag taagtgtaat
     1861 aaccgaagta gtttatgggc taccgaaata cacttacgcc agcattatga aagtgaatct
     1921 tacttttgta aaactttatg gtttgtggaa aacaaatgtt tttgaacatt taaaaagttc
     1981 agatgttaaa aagttgaaag gttaatgtaa aacaatcaat attaaagaat tttgatgcca
     2041 aaactattag ataaaaggtt aatctacatc cctactagaa ttctcatact taactggttg
     2101 gttatgtgga agaaacatac tttcacaata aagagcttta ggatatgatg ccattttata
     2161 tcactagtag gcagaccagc agactttttt ttattgtgat atgggataac ctaggcatac
     2221 tgcactgtac actctgacat atgaagtgct ctagtcaagt ttaactggtg tccacagagg
     2281 acatggttta actggaattc gtcaagcctc tggttctaat ttctcatttg caggaggacg
     2341 ctccagatga aatgggtaag tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc
     2401 agtaattcta ggcgatcgct cgagcccggg aattcgttta aacctagagc gtccgctggc
     2461 cgcaataaaa tatctttatt ttcattacac agctcactga gccatgcgct atgtcccgct
     2521 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat
     2581 ctaaatgagt cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg
     2641 cggggggagg gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg
     2701 gtggccacgg gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg
     2761 gaggcggggg tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc
     2821 gccccagagt gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg
     2881 cgacgggctc gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc
     2941 atccgcacga ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg
     3001 ctgacgcgga gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg
     3061 tcacccggag gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc
     3121 gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt
     3181 agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag
     3241 atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc
     3301 tcgaacaccg agcgaccctg cagcgacccg cttaaaagct tggcattccg gtactgttgg
     3361 taaagccacc atggccgatg ctaagaacat taagaagggc cctgctccct tctaccctct
     3421 ggaggatggc accgctggcg agcagctgca caaggccatg aagaggtatg ccctggtgcc
     3481 tggcaccatt gccttcaccg atgcccacat tgaggtggac atcacctatg ccgagtactt
     3541 cgagatgtct gtgcgcctgg ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg
     3601 catcgtggtg tgctctgaga actctctgca gttcttcatg ccagtgctgg gcgccctgtt
     3661 catcggagtg gccgtggccc ctgctaacga catttacaac gagcgcgagc tgctgaacag
     3721 catgggcatt tctcagccta ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct
     3781 gaacgtgcag aagaagctgc ctatcatcca gaagatcatc atcatggact ctaagaccga
     3841 ctaccagggc ttccagagca tgtacacatt cgtgacatct catctgcctc ctggcttcaa
     3901 cgagtacgac ttcgtgccag agtctttcga cagggacaaa accattgccc tgatcatgaa
     3961 cagctctggg tctaccggcc tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt
     4021 gcgcttctct cacgcccgcg accctatttt cggcaaccag atcatccccg acaccgctat
     4081 tctgagcgtg gtgccattcc accacggctt cggcatgttc accaccctgg gctacctgat
     4141 ttgcggcttt cgggtggtgc tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct
     4201 gcaagactac aaaattcagt ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa
     4261 gagcaccctg atcgacaagt acgacctgtc taacctgcac gagattgcct ctggcggcgc
     4321 cccactgtct aaggaggtgg gcgaagccgt ggccaagcgc tttcatctgc caggcatccg
     4381 ccagggctac ggcctgaccg agacaaccag cgccattctg attaccccag agggcgacga
     4441 caagcctggc gccgtgggca aggtggtgcc attcttcgag gccaaggtgg tggacctgga
     4501 caccggcaag accctgggag tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat
     4561 tatgtccggc tacgtgaata accctgaggc cacaaacgcc ctgatcgaca aggacggctg
     4621 gctgcactct ggcgacattg cctactggga cgaggacgag cacttcttca tcgtggaccg
     4681 cctgaagtct ctgatcaagt acaagggcta ccaggtggcc ccagccgagc tggagtctat
     4741 cctgctgcag caccctaaca ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc
     4801 cggcgagctg cctgccgccg tcgtcgtgct ggaacacggc aagaccatga ccgagaagga
     4861 gatcgtggac tatgtggcca gccaggtgac aaccgccaag aagctgcgcg gcggagtggt
     4921 gttcgtggac gaggtgccca agggcctgac cggcaagctg gacgcccgca agatccgcga
     4981 gatcctgatc aaggctaaga aaggcggcaa gatcgccgtg taataattct agagtcgggg
     5041 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     5101 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     5161 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     5221 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     5281 aatcgataag gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5341 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5401 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5461 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5521 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5581 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5641 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5701 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5761 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5821 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5881 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5941 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6001 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6061 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6121 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6181 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6241 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6301 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7081 cttttgctgg ccttttgctc acatggctcg ac
//
