LOCUS       sequence-419028-        6612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Oropouche orthobunyavirus NSm 
            protein from CMV promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6612)
  TITLE     Plasmids encoding N- and C-terminally HA tagged Oropouche and 
            Schmallenberg orthobunyavirus proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6612)
  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            "Unpublished"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: pCHA_OROV_NSm
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        347..1556
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1393..1412
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1560..2159
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1560..1579)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2056..2076
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2186..2234
                     /note="synthetic polyadenylation signal"
     CDS             2500..3294
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2500..3294
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2554..2573)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3164..3183
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3307..3443
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3588..3659
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3664..3710)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3728..3776
                     /note="synthetic polyadenylation signal"
     enhancer        3857..4160
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4162..4364
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4315..4335
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4972..4998
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5081..5202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5092..5111)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5146..5165
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5317..5363
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5393..5529
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5574..5645
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5871..6459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5951..5970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
       61 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
      121 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
      181 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
      241 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
      301 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg
      361 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa
      421 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
      481 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
      541 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
      601 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
      661 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
      721 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
      781 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
      841 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
      901 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
      961 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa
     1021 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca
     1081 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg
     1141 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc
     1201 tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc
     1261 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc
     1321 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc
     1381 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt
     1441 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt
     1501 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca
     1561 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     1621 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     1681 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     1741 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     1801 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     1861 cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg cccaaggagc
     1921 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     1981 gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg
     2041 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     2101 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg
     2161 ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt
     2221 tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg
     2281 cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac
     2341 ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg
     2401 tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca
     2461 aacatctatc aagagacagg atcagcagga ggctttcgca tgattgaaca agatggattg
     2521 cacgcaggtt ctccggcggc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2581 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg tccggttctt
     2641 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     2701 tcgtggctgg cgacgacggg cgttccttgc gcggctgtgc tcgacgttgt cactgaagcg
     2761 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     2821 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     2881 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     2941 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3001 gccgaactgt tcgccaggct caaggcgtct atgcccgacg gcgaggatct cgtcgtgacc
     3061 cacggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3121 gactgtggcc gtctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3181 attgctgaag agcttggcgg cgaatgggct gaccgcttcc ttgtgcttta cggtatcgcc
     3241 gcgcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaccgatt
     3301 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     3361 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     3421 ccttaccaga aatttatcct taaggtcgtt taaactcgac tctggctcta tcgaatctcc
     3481 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     3541 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     3601 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     3661 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     3721 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     3781 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     3841 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     3901 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     3961 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4021 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4081 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4141 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4201 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4261 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4321 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     4381 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgcatgcag
     4441 atgtgtaaat cgaaaagctg gtccttcaca gcagctatat taactggatt aatattgatg
     4501 gaattcatct caccaattgc aggagagaga atgtataagc ttgaggagct ggctgatgag
     4561 tttataaagc taacagagca ggtaaatatt ttagaaagag aaatggaagt cttgaagcag
     4621 tcaataattg ttatgtttgc aatcattttg gtattgttgc tttttgaaaa tatgatattc
     4681 aaccggcttt tccgcatttt ttatcgttct tgctccatgt gcggcttgat acattataga
     4741 ccaggactca agatcgattt aaccaaaact aacagatgtg gtagttgcat ctgtggattc
     4801 gacgagcagc aatcatctgg ttttgaatat gaaatatttc taaaggatat gcatgtccag
     4861 aaagaatcat gcaaatttag tccaagagta aaccacttta ggaatattaa agctttatta
     4921 tttgcacttg taatctgtgc tactatatat acagtttatg cactcgagta ctatccgtat
     4981 gatgtgccgg attatgcgta gcgctatcga tttctggatc cgcaggcctc tgctagcttg
     5041 actgactgag atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt
     5101 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     5161 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     5221 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     5281 tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta
     5341 aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc
     5401 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5461 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5521 tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct
     5581 ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct
     5641 ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac
     5701 ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga
     5761 tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca
     5821 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5881 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5941 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6001 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6061 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6121 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6181 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6241 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6301 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6361 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6421 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6481 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6541 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6601 agtatatatg ag
//
