LOCUS       sequence-393445-        6144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  siRNA vector expressing TMV P19 and a 425bp-long NP sequence as 
            inverted repeat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  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 6144)
  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: pBAD-P19-NP
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(226..244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        312..416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(417..485,486..1277)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(635..654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1319..1774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1406..1425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1616..1637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1885..2473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2374..2393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2627..2644
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2659..2799
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2725..2744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3045..3064
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3244..4122)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4149..4433
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4356..4375
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4460..4482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4499..4516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5039..5056
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     5063..5082
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5063..5081
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6126..6142)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6126..6142)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
       61 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
      121 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
      181 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
      241 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      301 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      361 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      421 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      481 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      541 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      601 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      661 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      721 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      781 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      841 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      901 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      961 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1021 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1081 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1141 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1201 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1261 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac
     1321 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1381 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1441 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1501 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     1561 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1621 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa
     1681 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1741 gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt
     1801 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1861 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1921 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1981 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2041 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2101 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2161 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2221 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2281 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2341 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2401 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2461 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2521 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2581 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2641 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2701 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2761 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2821 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2881 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2941 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     3001 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     3061 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     3121 aggatctgct catgtttgac agcttatcat cgatgcataa tgtgcctgtc aaatggacga
     3181 agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac
     3241 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
     3301 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
     3361 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
     3421 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     3481 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     3541 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     3601 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     3661 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     3721 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
     3781 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     3841 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     3901 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     3961 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     4021 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     4081 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     4141 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
     4201 ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag
     4261 cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt
     4321 ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag
     4381 attagcggat cctacctgac gctttttatc gcaactctct actgtttctc catacccgtt
     4441 tttttgggct agaaataatt ttgtttaact ttaagaagga gatatacata tggctagtca
     4501 tcaccatcac catcacagat ccatggaacg agctatacaa ggaaacgacg ctagggaaca
     4561 agctaacagt gaacgttggg atggaggatc aggaggtacc acttctccct tcaaacttcc
     4621 tgacgaaagt ccgagttgga ctgagtggcg gctacataac gatgagacga attcgaatca
     4681 agataatccc cttggtttca aggaaagctg gggtttcggg aaagttgtat ttaagagata
     4741 tctcagatac gacaggacgg aagcttcact gcacagagtc cttggatctt ggacgggaga
     4801 ttcggttaac tatgcagcat ctcgattttt cggtttcgac cagatcggat gtacctatag
     4861 tattcggttt cgaggagtta gtatcaccgt ttctggaggg tcgcgaactc ttcagcatct
     4921 ctgtgagatg gcaattcggt ctaagcaaga actgctacag cttgccccaa tcgaagtgga
     4981 aagtaatgta tcaagaggat gccctgaagg tactcaaacc ttcgaaaaag aaagcgagca
     5041 ccaccaccac caccactgaa tctaatacga ctcactatag gggagctact agtggatccc
     5101 cctgcttcaa atgaaaacat ggaaacaatg gactctatta ctcttgaact gagaagcaaa
     5161 tactgggcta taagaaccag gagcggagga aataccaatc aacagagagc atctgcaggg
     5221 caaatcagtg tacaacctac tttctcagta cagagagatc ttcctttcga gagagtgacc
     5281 atcatggcag catttacagg gaacactgaa ggcagaacat ctgacatgag gactgaaatt
     5341 ataagaatga tggaaagtgc cagaccagaa gatgtgtcct tccaggggcg gggagtcttc
     5401 gagctctcgg acgaaaaggc aacgaacccg atcgtgcctt cctttgacat gagtaatgag
     5461 ggatcttatt tcttcggaga caatgcagag gagtatgaca attaaagaaa aatacccttg
     5521 tttctactag ggctgcagga attcgatatc aagcttatcg gtcgatgaga tatttatgcc
     5581 agccagccag acgcagacgc gccgagacag aactttcgac cggtgctctc gtgtataaac
     5641 tctctgtcta gactcgacgg tatcgataag cttgatatcg aattcctgca gccctagtag
     5701 aaacaagggt atttttcttt aattgtcata ctcctctgca ttgtctccga agaaataaga
     5761 tccctcatta ctcatgtcaa aggaaggcac gatcgggttc gttgcctttt cgtccgagag
     5821 ctcgaagact ccccgcccct ggaaggacac atcttctggt ctggcacttt ccatcattct
     5881 tataatttca gtcctcatgt cagatgttct gccttcagtg ttccctgtaa atgctgccat
     5941 gatggtcact ctctcgaaag gaagatctct ctgtactgag aaagtaggtt gtacactgat
     6001 ttgccctgca gatgctctct gttgattggt atttcctccg ctcctggttc ttatagccca
     6061 gtatttgctt ctcagttcaa gagtaataga gtccattgtt tccatgtttt catttgaagc
     6121 agggggatcc actagttcta gagc
//
