LOCUS       sequence-381509-        7018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual-Luciferase Reporter vector carrying an EGFP direct sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  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 7018)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..7018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    19..67
                     /note="synthetic polyadenylation signal"
     promoter        76..827
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      857..866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             863..2515
                     /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             1822..1830
                     /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    2559..2680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2570..2589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2624..2643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2813..2917
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2918..2986,2987..3778)
                     /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(3136..3155)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3949..4537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4438..4457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4666..5023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4666..4857
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4689..4709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4874..5009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5093..5225
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5208..5227
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5270..5288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5292..5301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5298..6233
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             join(6260..6262,6263..6265,6266..6979)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6260..6262,6263..6265,6266..6976)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6305..6326)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6566..6585)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6913..6934
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tcgacggccg ctggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
       61 ttgtgtgagg atctaaatga gtcttcggac ctcgcggggg ccgcttaagc ggtggttagg
      121 gtttgtctga cgcgggggga gggggaagga acgaaacact ctcattcgga ggcggctcgg
      181 ggtttggtct tggtggccac gggcacgcag aagagcgccg cgatcctctt aagcaccccc
      241 ccgccctccg tggaggcggg ggtttggtcg gcgggtggta actggcgggc cgctgactcg
      301 ggcgggtcgc gcgccccaga gtgtgacctt ttcggtctgc tcgcagaccc ccgggcggcg
      361 ccgccgcggc ggcgacgggc tcgctgggtc ctaggctcca tggggaccgt atacgtggac
      421 aggctctgga gcatccgcac gactgcggtg atattaccgg agaccttctg cgggacgagc
      481 cgggtcacgc ggctgacgcg gagcgtccgt tgggcgacaa acaccaggac ggggcacagg
      541 tacactatct tgtcacccgg aggcgcgagg gactgcagga gcttcaggga gtggcgcagc
      601 tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat
      661 ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc attggcgaat
      721 tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg
      781 acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaaaag cttggcattc
      841 cggtactgtt ggtaaagcca ccatggccga tgctaagaac attaagaagg gccctgctcc
      901 cttctaccct ctggaggatg gcaccgctgg cgagcagctg cacaaggcca tgaagaggta
      961 tgccctggtg cctggcacca ttgccttcac cgatgcccac attgaggtgg acatcaccta
     1021 tgccgagtac ttcgagatgt ctgtgcgcct ggccgaggcc atgaagaggt acggcctgaa
     1081 caccaaccac cgcatcgtgg tgtgctctga gaactctctg cagttcttca tgccagtgct
     1141 gggcgccctg ttcatcggag tggccgtggc ccctgctaac gacatttaca acgagcgcga
     1201 gctgctgaac agcatgggca tttctcagcc taccgtggtg ttcgtgtcta agaagggcct
     1261 gcagaagatc ctgaacgtgc agaagaagct gcctatcatc cagaagatca tcatcatgga
     1321 ctctaagacc gactaccagg gcttccagag catgtacaca ttcgtgacat ctcatctgcc
     1381 tcctggcttc aacgagtacg acttcgtgcc agagtctttc gacagggaca aaaccattgc
     1441 cctgatcatg aacagctctg ggtctaccgg cctgcctaag ggcgtggccc tgcctcatcg
     1501 caccgcctgt gtgcgcttct ctcacgcccg cgaccctatt ttcggcaacc agatcatccc
     1561 cgacaccgct attctgagcg tggtgccatt ccaccacggc ttcggcatgt tcaccaccct
     1621 gggctacctg atttgcggct ttcgggtggt gctgatgtac cgcttcgagg aggagctgtt
     1681 cctgcgcagc ctgcaagact acaaaattca gtctgccctg ctggtgccaa ccctgttcag
     1741 cttcttcgct aagagcaccc tgatcgacaa gtacgacctg tctaacctgc acgagattgc
     1801 ctctggcggc gccccactgt ctaaggaggt gggcgaagcc gtggccaagc gctttcatct
     1861 gccaggcatc cgccagggct acggcctgac cgagacaacc agcgccattc tgattacccc
     1921 agagggcgac gacaagcctg gcgccgtggg caaggtggtg ccattcttcg aggccaaggt
     1981 ggtggacctg gacaccggca agaccctggg agtgaaccag cgcggcgagc tgtgtgtgcg
     2041 cggccctatg attatgtccg gctacgtgaa taaccctgag gccacaaacg ccctgatcga
     2101 caaggacggc tggctgcact ctggcgacat tgcctactgg gacgaggacg agcacttctt
     2161 catcgtggac cgcctgaagt ctctgatcaa gtacaagggc taccaggtgg ccccagccga
     2221 gctggagtct atcctgctgc agcaccctaa cattttcgac gccggagtgg ccggcctgcc
     2281 cgacgacgat gccggcgagc tgcctgccgc cgtcgtcgtg ctggaacacg gcaagaccat
     2341 gaccgagaag gagatcgtgg actatgtggc cagccaggtg acaaccgcca agaagctgcg
     2401 cggcggagtg gtgttcgtgg acgaggtgcc caagggcctg accggcaagc tggacgcccg
     2461 caagatccgc gagatcctga tcaaggctaa gaaaggcggc aagatcgccg tgtaataatt
     2521 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2581 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2641 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2701 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2761 caaatgtggt aaaatcgata aggatccagg tggcactttt cggggaaatg tgcgcggaac
     2821 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2881 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2941 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3001 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3061 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3121 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3181 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3241 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3301 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3361 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3421 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3481 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3541 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3601 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3661 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3721 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3781 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4081 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4561 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct gcgcagcacc
     4621 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     4681 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4741 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4861 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4921 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4981 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga
     5041 cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat
     5101 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     5161 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     5221 cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc
     5281 actataggct agccaccatg gcttccaagg tgtacgaccc cgagcaacgc aaacgcatga
     5341 tcactgggcc tcagtggtgg gctcgctgca agcaaatgaa cgtgctggac tccttcatca
     5401 actactatga ttccgagaag cacgccgaga acgccgtgat ttttctgcat ggtaacgctg
     5461 cctccagcta cctgtggagg cacgtcgtgc ctcacatcga gcccgtggct agatgcatca
     5521 tccctgatct gatcggaatg ggtaagtccg gcaagagcgg gaatggctca tatcgcctcc
     5581 tggatcacta caagtacctc accgcttggt tcgagctgct gaaccttcca aagaaaatca
     5641 tctttgtggg ccacgactgg ggggcttgtc tggcctttca ctactcctac gagcaccaag
     5701 acaagatcaa ggccatcgtc catgctgaga gtgtcgtgga cgtgatcgag tcctgggacg
     5761 agtggcctga catcgaggag gatatcgccc tgatcaagag cgaagagggc gagaaaatgg
     5821 tgcttgagaa taacttcttc gtcgagacca tgctcccaag caagatcatg cggaaactgg
     5881 agcctgagga gttcgctgcc tacctggagc cattcaagga gaagggcgag gttagacggc
     5941 ctaccctctc ctggcctcgc gagatccctc tcgttaaggg aggcaagccc gacgtcgtcc
     6001 agattgtccg caactacaac gcctaccttc gggccagcga cgatctgcct aagatgttca
     6061 tcgagtccga ccctgggttc ttttccaacg ctattgtcga gggagctaag aagttcccta
     6121 acaccgagtt cgtgaaggtg aagggcctcc acttcagcca ggaggacgct ccagatgaaa
     6181 tgggtaagta catcaagagc ttcgtggagc gcgtgctgaa gaacgagcag taattctagg
     6241 cgatcgctcg agcggccgca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     6301 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     6361 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     6421 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     6481 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     6541 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     6601 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     6661 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     6721 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     6781 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     6841 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     6901 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     6961 ggacgagctg tacaagtagn nnnnngctga tcagcgggtt taaacgggcc ctctagac
//
