LOCUS       sequence-389054-        6263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promega's psiCHECK-2 Dual-Luciferase Reporter vector modified to 
            allow the insertion of the target in opposite directions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  AUTHORS   Villa R, Renzi S, Dotti S, Lucchini F
  TITLE     siRNAs pools generated in Escherichia coli exhibit strong 
            RNA-interference activity against influenza virus genomic sequences.
  JOURNAL   Virology. 2022 Dec 31;579:38-45. doi: 10.1016/j.virol.2022.12.013.
  PUBMED    36599198
REFERENCE   2  (bases 1 to 6263)
  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: "Virology.";
            date: "2022-12-31"; volume: "579:38-45. doi"; pages: " 
            10.1016/j.virol.2022.12.013"
COMMENT     Sequence Label: psiCHECK-2-XNS
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        61..252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          488..620
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     603..622
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        665..683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1628
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1677..1725
                     /note="synthetic polyadenylation signal"
     promoter        1734..2485
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2515..2524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2521..4173
                     /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             3480..3488
                     /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    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4282..4301
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4471..4575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4576..4644,4645..5436)
                     /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(4794..4813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5607..6195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6096..6115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
       61 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      481 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
      541 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
      601 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
      661 tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac gaccccgagc
      721 aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc
      781 tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc
      841 tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg
      901 tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg
      961 gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc
     1021 ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact
     1081 cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga
     1141 tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag
     1201 agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga
     1261 tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg
     1321 gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca
     1381 agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc
     1441 tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag
     1501 ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg
     1561 acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg
     1621 agcagtaatt ctaggcgatc gctcgagcgg ccgcatggtc gacggccgct ggccgcaata
     1681 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt
     1741 cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg
     1801 gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg
     1861 gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg
     1921 tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt
     1981 gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc
     2041 gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga
     2101 ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga
     2161 gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag
     2221 gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt
     2281 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
     2341 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
     2401 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
     2461 agcgaccctg cagcgacccg cttaaaagct tggcattccg gtactgttgg taaagccacc
     2521 atggccgatg ctaagaacat taagaagggc cctgctccct tctaccctct ggaggatggc
     2581 accgctggcg agcagctgca caaggccatg aagaggtatg ccctggtgcc tggcaccatt
     2641 gccttcaccg atgcccacat tgaggtggac atcacctatg ccgagtactt cgagatgtct
     2701 gtgcgcctgg ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg
     2761 tgctctgaga actctctgca gttcttcatg ccagtgctgg gcgccctgtt catcggagtg
     2821 gccgtggccc ctgctaacga catttacaac gagcgcgagc tgctgaacag catgggcatt
     2881 tctcagccta ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct gaacgtgcag
     2941 aagaagctgc ctatcatcca gaagatcatc atcatggact ctaagaccga ctaccagggc
     3001 ttccagagca tgtacacatt cgtgacatct catctgcctc ctggcttcaa cgagtacgac
     3061 ttcgtgccag agtctttcga cagggacaaa accattgccc tgatcatgaa cagctctggg
     3121 tctaccggcc tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct
     3181 cacgcccgcg accctatttt cggcaaccag atcatccccg acaccgctat tctgagcgtg
     3241 gtgccattcc accacggctt cggcatgttc accaccctgg gctacctgat ttgcggcttt
     3301 cgggtggtgc tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct gcaagactac
     3361 aaaattcagt ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg
     3421 atcgacaagt acgacctgtc taacctgcac gagattgcct ctggcggcgc cccactgtct
     3481 aaggaggtgg gcgaagccgt ggccaagcgc tttcatctgc caggcatccg ccagggctac
     3541 ggcctgaccg agacaaccag cgccattctg attaccccag agggcgacga caagcctggc
     3601 gccgtgggca aggtggtgcc attcttcgag gccaaggtgg tggacctgga caccggcaag
     3661 accctgggag tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc
     3721 tacgtgaata accctgaggc cacaaacgcc ctgatcgaca aggacggctg gctgcactct
     3781 ggcgacattg cctactggga cgaggacgag cacttcttca tcgtggaccg cctgaagtct
     3841 ctgatcaagt acaagggcta ccaggtggcc ccagccgagc tggagtctat cctgctgcag
     3901 caccctaaca ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc cggcgagctg
     3961 cctgccgccg tcgtcgtgct ggaacacggc aagaccatga ccgagaagga gatcgtggac
     4021 tatgtggcca gccaggtgac aaccgccaag aagctgcgcg gcggagtggt gttcgtggac
     4081 gaggtgccca agggcctgac cggcaagctg gacgcccgca agatccgcga gatcctgatc
     4141 aaggctaaga aaggcggcaa gatcgccgtg taataattct agagtcgggg cggccggccg
     4201 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4261 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4321 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4381 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4441 gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4561 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4681 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4741 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4801 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4861 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4981 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5101 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5161 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5281 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5401 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6241 ccttttgctc acatggctcg aca
//
