LOCUS       sequence-473233-        3396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding Oropouche virus (OROV) nucleoprotein (N) with an 
            N-terminal His6 and biotin acceptor peptide (BAP, a.k.a. AviTag) for
            expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3396)
  AUTHORS   Merchant MK, de Paula Souza J, Abdelkarim S, Chakraborty S, Nasser 
            Neto TA, Gutierrez Manchay KI, de Melo Jorge DM, do Nascimento VA, 
            Sun Y, Caroe ER, Eyssen LEA, Rudd JS, Ribeiro Piauilino IC, 
            Damasceno Pinto S, Pereira da Silva MH, Rocha do Nascimento F, 
            Naveca FG, Proenca-Modena JL, da Silva LLP, Pinto de Figueiredo RM, 
            Owens RJ, Arruda E, Graham SC
  TITLE     Protein-based tools for the detection and characterisation of 
            Oropouche virus infection.
  JOURNAL   EMBO Mol Med. 2025 Aug 11. doi: 10.1038/s44321-025-00291-7.
  PUBMED    40790101
REFERENCE   2  (bases 1 to 3396)
  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: "EMBO Mol 
            Med. 2025 Aug 11. doi: 10.1038/s44321-025-00291-7."
COMMENT     Sequence Label: pMWCAVI_H6BAP-OROV_N
FEATURES             Location/Qualifiers
     source          1..3396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     347..364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    379..518
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(444..463)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     604..626
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     679..698
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        679..697
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             729..751
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             759..776
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             777..800
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             804..848
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1572..1607
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     primer_bind     complement(1673..1691)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1687..1734
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1779..1797)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1865..1969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1970..2038,2039..2830)
                     /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(2188..2207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3001..3396,1..193)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
       61 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      121 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      181 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      241 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      301 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      361 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      421 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
      481 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
      541 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
      601 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagg
      661 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
      721 gaaataattt tgtttaactt taagaaggag atataatgca tcaccatcac caccatctgg
      781 aagtgctgtt tcagggcccg agcggcctga acgatatttt tgaagcgcag aaaattgaat
      841 ggcatgaaca tatgagcgag ttcatcttca acgacgtgcc ccagcggacc accagcacct
      901 ttgatcctga agccgcctac gtggccttcg aggccagata tggacaggtg ctgaatgccg
      961 gcgttgtgcg ggtgttcttc ctgaaccaga aaaaggccaa ggacgtgctg agaaagacca
     1021 gcagacccat ggtggacctg acctttggcg gagtgcagtt cgccatggtc aacaatcact
     1081 tccctcagtt ccagagcaac cccgtgcctg acaatggcct gacactgcac agactgagcg
     1141 gctaccttgc cagatgggcc ttcacacaga tgcggagccc tatcaagcag gccgagttta
     1201 gagccaccgt ggtggtgcct ctggccgaag tgaaaggctg cacctggaat gatggcgacg
     1261 ccatgtatct gggctttgcc gctggcgccg agatgttcct gcagaccttc accttctttc
     1321 cactggtcat cgagatgcac cgggtgctga aggacggcat ggacgtgaac tttatgaaga
     1381 aggtgctgcg gcagcgctac ggccagaaaa cagctgagca gtggatgcgg gaagagatcg
     1441 tggccgttag agccgccttt gaggctgtgg gaacactggc ctgggccaga acaggcttta
     1501 gccctgccgc cagagacttc ctgagacagt tcggcatcga catctgagga tccccaggaa
     1561 ttgcgagcat gccggatcgc aaagcggcgg tgagccattg gcagcagcgc agctaagaat
     1621 tccgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1681 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1741 ggaggaacta tatccggata attcttgaag acgaaagggc ctcgtgatac gcctattttt
     1801 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1921 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1981 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2041 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2101 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2161 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     2221 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2281 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2341 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2401 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2461 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     2521 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2581 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2641 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2701 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2761 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2821 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3361 ctacaccgaa ctgagatacc tacagcgtga gctatg
//
