LOCUS       sequence-418962-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally HA-tagged Oropouche orthobunyavirus Gc 
            protein from CMV promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  TITLE     Plasmids encoding N- and C-terminally HA tagged Oropouche and 
            Schmallenberg orthobunyavirus proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6290)
  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            "Unpublished"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_Gc
FEATURES             Location/Qualifiers
     source          1..6290
                     /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    3429..3563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3453..3472)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3507..3526
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3674..3690)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3674..3690)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3687..3709)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3698..3714
                     /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(3722..3728,3729..3746,3747..3752))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3767..3788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3905..3922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4076..4664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4156..4175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4835..5626,5627..5695))
                     /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     5458..5477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5696..5800)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5868..5886
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5924..5946)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6046..6065
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6259..6281
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6273..6290
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6274..6290
                     /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 gaggaattcc tcgagtattc tgatttattc
      901 tcaaaaaatc tagtcacaga atatgataaa aaatattttg aacctgatac tgtaaatgat
      961 caattcaaca agatagagtt tggccaagat gcacatagaa tgatactatt agaaagaata
     1021 ctttacaaga cagaatgtga aatgctaagc ttaaaaaaga acagtggacc ttataatgtt
     1081 gcctggcgca catattttaa aaatcataat atagacttat gcagtaggca caactacaaa
     1141 atgatatgtc agtgtattaa tacacactct atgtgtaaga atacagatat agattataac
     1201 aaagagatcg aaacatatta taaaagcaat gctgcagcct atagagctga cctgaacacc
     1261 ataatggaca cattaaaaac agcattcaga gggcttacaa aggtcctaat tgaaaattat
     1321 atagaaaaag atgactctga tgctcttaag gccttattta gtaatattac tgattctgtt
     1381 caggacaatt atcagatgat tggaatattg aaatttgcca gtaagctatt agatattaac
     1441 cttggtagat ccacccgctc agctcatcat tctataatga caaacgaaat tcccaaatct
     1501 aatcctttta ctgattacag ttattctaat ctgaatataa aggagtgcat gtcaccagag
     1561 agcttgaaat gttttaagaa gagagacagt acaccgcata caaaccacct gctttgcaag
     1621 atagacaaca aatataaagc atttgactgg ccagagatag aaacaattca aaaaggtcaa
     1681 aaattgtgcc ttggtgacag tcattgcaac ttggaattta cagccataac tgcagacaag
     1741 attatgtcat taactaactg ttataaagaa tctttcactg cgcaaccagc agatatgcag
     1801 gctggaatta aaaaatgctc agctgatgaa attggagaat gtacaacact tgaagacaag
     1861 acatggccaa ttatattctg tggtggcaaa tattactact ctgatagcaa ggagcatgcc
     1921 aaggacggtt ctatcaacaa ttattgttta accaacaaat gctctgaaca acgtttccca
     1981 atacatgaaa attggttcaa aaaatgtaat tgggacaaga cccacaaaga atttagcact
     2041 atgagacaaa taaactataa tgatataact tcttatagaa aggcaataga atctgagata
     2101 ggaacagatt tgatgacaca ccattataag cccacaaaga accttccaca tgttgtacca
     2161 aggtatcata gtatagatgt acaaggtaca gaatcaacag agggtattat caatggtttc
     2221 atacaaaata caatacctgc tattagtggt ctaggtgtcg gttatcatct gaatttcaaa
     2281 ggcaatcagt tatttgacat agttattttt gtgaaaaaag ctgtatataa agcacagtac
     2341 cagaaagcat acaccacagg tcctagcata tccataaata ttgaacacaa cgaaagatgc
     2401 acaggccact gccccgaaaa gattcctgca aaagaaggtt ggttaacatt ttctaaagag
     2461 catactagtt cttggggatg tgaagaatat ggctgtcttg ctatagatac aggctgtcta
     2521 tatggctctt gtcaagatgt catacgccca gaattagatg tttataagaa gattggcagt
     2581 gaggtatcac tgatagaaat atgtataact ttgcctcatg aaacttattg caatgatatg
     2641 gacatattag aaccgattat aggagataaa ttaagtgcat cctttcagaa tacacagact
     2701 aatcaacttc cgacccttat agcttataaa aaagggaaga tatacacagg tcaaatcaac
     2761 gatattggaa acacagcatt gcagtgcggc tcaatccaag taatcaatgg atcaactatc
     2821 ggaactggca gccccaaatt tgattatata tgccatgcta tgagaaggaa agatgttatt
     2881 gtaaggaagt gtttcaatga caattatcag tcgtgcacta ggctagaaaa acgaaacgac
     2941 ttgatccctt atagaaaagg cgatgtcatc gaagtatcaa aaacaggatc gaacatgggc
     3001 caaatgacat ttaaaataga actaggtgat atcaactata aaatatttac taaatcaata
     3061 gatcttcaaa tgtctggcat ttgtgctggg tgcattgact gtgctgaagg catttcatgc
     3121 tctataaatg ctgaagtgcc agccgaaaca gtctgccatt gcaaaacaaa ctgtgaagat
     3181 tttataaaca atatagtgtt ttcaccgcaa atcaagaatt acaatataaa agtccactgt
     3241 aaatctaaag ttgaaaaaat aacagcacat atctgtggga gagacattga tttacagtta
     3301 acaattaagc catacaatca gaagatagac ttgtcgcagt tagatgaatc aaattacata
     3361 agggaagagg acctccagtg tggaacatgg ttatgtaaag tgcaaaaaga ggggtaggcg
     3421 gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3481 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3541 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3601 agggggaggt gtgggaggtt ttttcggatc ctctagagtc gatctgcagg catgctagct
     3661 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3721 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3781 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3841 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3901 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3961 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4021 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4081 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4141 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4201 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4261 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4321 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4381 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4441 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4501 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4561 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4621 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4681 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4741 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4801 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4861 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4921 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4981 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5041 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5101 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5161 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5221 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5281 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5341 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5401 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5461 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5521 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5581 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5641 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5701 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5761 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5821 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5881 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5941 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6001 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6061 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6121 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6181 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     6241 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
//
