LOCUS       sequence-389051-        5396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from pGLO (BioRad) by the insertion of a 6xHis Tag and a 
            BglII site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5396)
  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 5396)
  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-6xHis-BglII
FEATURES             Location/Qualifiers
     source          1..5396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3..716
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(30..58)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     653..673
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    718..774
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(807..824)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      977..1063
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1155..1182
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1201..1292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1293..1361,1362..2153)
                     /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(1511..1530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2195..2650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2282..2301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2492..2513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2761..3349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3250..3269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3503..3520
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3535..3675
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3601..3620)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3761..3783
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3921..3940
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4120..4998)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5025..5309
                     /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     5232..5251
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             5336..5358
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5375..5392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctagcaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
       61 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgct acatacggaa
      121 agcttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
      181 tcactacttt ctcttatggt gttcaatgct tttcccgtta tccggatcat atgaaacggc
      241 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact atatctttca
      301 aagatgacgg gaactacaag acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta
      361 atcgtatcga gttaaaaggt attgatttta aagaagatgg aaacattctc ggacacaaac
      421 tcgagtacaa ctataactca cacaatgtat acatcacggc agacaaacaa aagaatggaa
      481 tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg atccgttcaa ctagcagacc
      541 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
      601 tgtcgacaca atctgccctt tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc
      661 ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac aaataatgaa
      721 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggctgt
      781 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
      841 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
      901 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
      961 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1021 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1081 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1141 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1261 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1321 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1381 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1441 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1501 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1561 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1621 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1681 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1741 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1801 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     1861 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1921 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1981 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2041 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2101 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2161 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     2221 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2281 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2341 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2401 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2461 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2521 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2581 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2641 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2881 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3001 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3061 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3421 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3481 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3541 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     3601 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3661 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3721 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3781 aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc caacagtccc
     3841 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     3901 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     3961 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg tttgacagct
     4021 tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg
     4081 ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac
     4141 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     4201 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     4261 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     4321 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     4381 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     4441 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     4501 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     4561 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     4621 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     4681 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     4741 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     4801 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     4861 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     4921 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     4981 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     5041 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     5101 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     5161 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     5221 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     5281 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagaa ataattttgt
     5341 ttaactttaa gaaggagata tacatatggc tagtcatcac catcaccatc acagat
//
