LOCUS       sequence-418963-        4457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally HA-tagged Oropouche orthobunyavirus 
            nucleocapsid protein from CMV promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4457)
  TITLE     Plasmids encoding N- and C-terminally HA tagged Oropouche and 
            Schmallenberg orthobunyavirus proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4457)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pNHA_OROV_NCP
FEATURES             Location/Qualifiers
     source          1..4457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             832..858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1596..1730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1620..1639)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1674..1693
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1841..1857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1841..1857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1854..1876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1865..1881
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1889..1895,1896..1913,1914..1919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1934..1955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2072..2089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2243..2831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2323..2342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3002..3793,3794..3862))
                     /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     3625..3644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3863..3967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4035..4053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4091..4113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4213..4232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4426..4448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4440..4457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4441..4457
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat gtacccatac
      841 gatgttccag attacgctct tatggccatg gaggaattct cagagttcat tttcaacgat
      901 gtaccacaac ggactacatc tacatttgat ccggaggcag catatgtggc atttgaagct
      961 agatacggac aagtgctcaa tgctggtgtt gttagagtct tcttcctcaa ccaaaagaag
     1021 gccaaagatg tcttacgtaa gacatcgagg cccatggttg accttacttt tggtggggtc
     1081 caatttgcaa tggttaataa ccatttccca cagttccagt cgaatccagt gccggacaac
     1141 ggtcttaccc tgcaccgtct gtcaggatac ctagcgcgct gggcctttac ccagatgcga
     1201 tcaccaatta agcaagctga gttcagagcc actgtagtag tgcctttggc tgaggtaaag
     1261 ggctgtactt ggaatgatgg tgacgcaatg tacctggggt ttgctgctgg tgctgagatg
     1321 ttcctgcaaa cattcacttt cttccctttg gtgattgaga tgcatagggt tctcaaggat
     1381 ggcatggatg tcaactttat gaagaaagtc ctccggcaac ggtatggcca aaagactgcc
     1441 gagcaatgga tgcgtgaaga aatagttgct gtaagagcag cttttgaagc tgtaggcact
     1501 ctggcctggg ccagaactgg attctcccca gcagcaagag acttcttgcg ccaattcgga
     1561 attgacatat agctcgagga tatcgcggcc gcggggatcc agacatgata agatacattg
     1621 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1681 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1741 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc
     1801 tagagtcgat ctgcaggcat gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1861 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1921 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4021 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     4081 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4141 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     4201 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     4261 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     4321 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     4381 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     4441 gtaaaacgac ggccagt
//
