LOCUS       sequence-389052-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual-Luciferase Reporter vector carrying a 425bp-long Influenza 
            virus NP sequence in forward direction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  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 6733)
  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-NP
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      4..13
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10..945
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     complement(1432..1448)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1432..1448)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    1464..1512
                     /note="synthetic polyadenylation signal"
     promoter        1521..2272
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2302..2311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2308..3960
                     /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             3267..3275
                     /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    4004..4125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4015..4034)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4069..4088
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4258..4362
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4363..4431,4432..5223)
                     /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(4581..4600)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5394..5982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5883..5902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6111..6468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6111..6302
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6134..6154)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6319..6454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          6538..6670
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6653..6672
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        6715..6733
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctagccacca tggcttccaa ggtgtacgac cccgagcaac gcaaacgcat gatcactggg
       61 cctcagtggt gggctcgctg caagcaaatg aacgtgctgg actccttcat caactactat
      121 gattccgaga agcacgccga gaacgccgtg atttttctgc atggtaacgc tgcctccagc
      181 tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg ctagatgcat catccctgat
      241 ctgatcggaa tgggtaagtc cggcaagagc gggaatggct catatcgcct cctggatcac
      301 tacaagtacc tcaccgcttg gttcgagctg ctgaaccttc caaagaaaat catctttgtg
      361 ggccacgact ggggggcttg tctggccttt cactactcct acgagcacca agacaagatc
      421 aaggccatcg tccatgctga gagtgtcgtg gacgtgatcg agtcctggga cgagtggcct
      481 gacatcgagg aggatatcgc cctgatcaag agcgaagagg gcgagaaaat ggtgcttgag
      541 aataacttct tcgtcgagac catgctccca agcaagatca tgcggaaact ggagcctgag
      601 gagttcgctg cctacctgga gccattcaag gagaagggcg aggttagacg gcctaccctc
      661 tcctggcctc gcgagatccc tctcgttaag ggaggcaagc ccgacgtcgt ccagattgtc
      721 cgcaactaca acgcctacct tcgggccagc gacgatctgc ctaagatgtt catcgagtcc
      781 gaccctgggt tcttttccaa cgctattgtc gagggagcta agaagttccc taacaccgag
      841 ttcgtgaagg tgaagggcct ccacttcagc caggaggacg ctccagatga aatgggtaag
      901 tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc agtaattcta ggcgatcgct
      961 cgacggtatc gataagcttg atatcgaatt cctgcagccc tagtagaaac aagggtattt
     1021 ttctttaatt gtcatactcc tctgcattgt ctccgaagaa ataagatccc tcattactca
     1081 tgtcaaagga aggcacgatc gggttcgttg ccttttcgtc cgagagctcg aagactcccc
     1141 gcccctggaa ggacacatct tctggtctgg cactttccat cattcttata atttcagtcc
     1201 tcatgtcaga tgttctgcct tcagtgttcc ctgtaaatgc tgccatgatg gtcactctct
     1261 cgaaaggaag atctctctgt actgagaaag taggttgtac actgatttgc cctgcagatg
     1321 ctctctgttg attggtattt cctccgctcc tggttcttat agcccagtat ttgcttctca
     1381 gttcaagagt aatagagtcc attgtttcca tgttttcatt tgaagcaggg ggatccacta
     1441 gttctagagc ggccgctggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     1501 gttttttgtg tgaggatcta aatgagtctt cggacctcgc gggggccgct taagcggtgg
     1561 ttagggtttg tctgacgcgg ggggaggggg aaggaacgaa acactctcat tcggaggcgg
     1621 ctcggggttt ggtcttggtg gccacgggca cgcagaagag cgccgcgatc ctcttaagca
     1681 cccccccgcc ctccgtggag gcgggggttt ggtcggcggg tggtaactgg cgggccgctg
     1741 actcgggcgg gtcgcgcgcc ccagagtgtg accttttcgg tctgctcgca gacccccggg
     1801 cggcgccgcc gcggcggcga cgggctcgct gggtcctagg ctccatgggg accgtatacg
     1861 tggacaggct ctggagcatc cgcacgactg cggtgatatt accggagacc ttctgcggga
     1921 cgagccgggt cacgcggctg acgcggagcg tccgttgggc gacaaacacc aggacggggc
     1981 acaggtacac tatcttgtca cccggaggcg cgagggactg caggagcttc agggagtggc
     2041 gcagctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa
     2101 tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg
     2161 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta
     2221 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aaaagcttgg
     2281 cattccggta ctgttggtaa agccaccatg gccgatgcta agaacattaa gaagggccct
     2341 gctcccttct accctctgga ggatggcacc gctggcgagc agctgcacaa ggccatgaag
     2401 aggtatgccc tggtgcctgg caccattgcc ttcaccgatg cccacattga ggtggacatc
     2461 acctatgccg agtacttcga gatgtctgtg cgcctggccg aggccatgaa gaggtacggc
     2521 ctgaacacca accaccgcat cgtggtgtgc tctgagaact ctctgcagtt cttcatgcca
     2581 gtgctgggcg ccctgttcat cggagtggcc gtggcccctg ctaacgacat ttacaacgag
     2641 cgcgagctgc tgaacagcat gggcatttct cagcctaccg tggtgttcgt gtctaagaag
     2701 ggcctgcaga agatcctgaa cgtgcagaag aagctgccta tcatccagaa gatcatcatc
     2761 atggactcta agaccgacta ccagggcttc cagagcatgt acacattcgt gacatctcat
     2821 ctgcctcctg gcttcaacga gtacgacttc gtgccagagt ctttcgacag ggacaaaacc
     2881 attgccctga tcatgaacag ctctgggtct accggcctgc ctaagggcgt ggccctgcct
     2941 catcgcaccg cctgtgtgcg cttctctcac gcccgcgacc ctattttcgg caaccagatc
     3001 atccccgaca ccgctattct gagcgtggtg ccattccacc acggcttcgg catgttcacc
     3061 accctgggct acctgatttg cggctttcgg gtggtgctga tgtaccgctt cgaggaggag
     3121 ctgttcctgc gcagcctgca agactacaaa attcagtctg ccctgctggt gccaaccctg
     3181 ttcagcttct tcgctaagag caccctgatc gacaagtacg acctgtctaa cctgcacgag
     3241 attgcctctg gcggcgcccc actgtctaag gaggtgggcg aagccgtggc caagcgcttt
     3301 catctgccag gcatccgcca gggctacggc ctgaccgaga caaccagcgc cattctgatt
     3361 accccagagg gcgacgacaa gcctggcgcc gtgggcaagg tggtgccatt cttcgaggcc
     3421 aaggtggtgg acctggacac cggcaagacc ctgggagtga accagcgcgg cgagctgtgt
     3481 gtgcgcggcc ctatgattat gtccggctac gtgaataacc ctgaggccac aaacgccctg
     3541 atcgacaagg acggctggct gcactctggc gacattgcct actgggacga ggacgagcac
     3601 ttcttcatcg tggaccgcct gaagtctctg atcaagtaca agggctacca ggtggcccca
     3661 gccgagctgg agtctatcct gctgcagcac cctaacattt tcgacgccgg agtggccggc
     3721 ctgcccgacg acgatgccgg cgagctgcct gccgccgtcg tcgtgctgga acacggcaag
     3781 accatgaccg agaaggagat cgtggactat gtggccagcc aggtgacaac cgccaagaag
     3841 ctgcgcggcg gagtggtgtt cgtggacgag gtgcccaagg gcctgaccgg caagctggac
     3901 gcccgcaaga tccgcgagat cctgatcaag gctaagaaag gcggcaagat cgccgtgtaa
     3961 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     4021 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4081 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4141 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     4201 ctctacaaat gtggtaaaat cgataaggat ccaggtggca cttttcgggg aaatgtgcgc
     4261 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4321 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4381 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4441 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4501 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4561 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4621 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4681 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4741 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4801 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4861 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4921 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4981 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5041 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5101 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5161 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5221 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5281 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5341 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5401 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5461 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5521 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5581 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5641 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5701 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5761 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5821 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5881 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5941 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6001 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatctgcgca
     6061 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     6121 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
     6481 tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac tgggcaggta
     6541 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     6601 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     6661 ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac
     6721 gactcactat agg
//
