LOCUS       sequence-419026-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Schmallenberg orthobunyavirus NSs 
            protein from CMV promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  TITLE     Plasmids encoding N- and C-terminally HA tagged Oropouche and 
            Schmallenberg orthobunyavirus proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6357)
  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            "Unpublished"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_SBV_NSs
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      154..742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     643..662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    968..1039
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1084..1220)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1250..1296)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1411..1532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1448..1467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1502..1521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1615..1641)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1994..2196)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2023..2043)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2198..2501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2582..2630
                     /note="synthetic polyadenylation signal"
     terminator      2648..2694
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2699..2770
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2915..3051)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(3064..3858)
                     /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             complement(3064..3858)
                     /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(3175..3194)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3785..3804
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     polyA_signal    4124..4172
                     /note="synthetic polyadenylation signal"
     CDS             complement(4199..4798)
                     /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(4282..4302)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4779..4798
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4802..6011)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4946..4965)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
       61 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      301 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      781 ttgctggcct tttgctcaca tgatttaaat gaaactcgta atcggagaac gggtgcaata
      841 gcaaggacca gatcgaactt gactgacgca attctgagcg tagtcgaaac ccagacaagt
      901 cgatctcgcc aagtagctga cttgactggt taattaagtt tcgatagccc aaggtaacca
      961 atgagattag ggagggaaga aagtttgatg aactacttct gacccctagt ggtgtccaga
     1021 aaagaccatt aaaggaatgt cttgatcgaa atcgagagat cgcctgcagg atagctgacg
     1081 accttaagga taaatttctg gtaaggagga cacgtatgga agtgggcaag ttggggaagc
     1141 cgtatccgtt gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc
     1201 atttttttct gcgccaatgc cggtagatag caatccggcc cgaggggcac aaaaaacccc
     1261 tcaagacccg tttagaggcc ccaaggggtt atgctacgat accgatagag atgggccaat
     1321 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1381 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1441 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1501 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtgagctgaa ggtacgctgt
     1561 atctcagtca gtcaagctag cagaggcctg cggatccaga aatcgatagc gctacgcata
     1621 atccggcaca tcatacggat agtactcgag gatatcctga catcctgtga agtgtaatgg
     1681 cattgtctgg cacaggattt gagacatatt ggggaaaatg gttattaacc actgtaaatt
     1741 tgaccccacc aaaagtaaga tcgacacttg gttgtgccgt cttatgtagg accatcttgg
     1801 ccttcttctg gttgaggaag aagactctag caacaccgaa gttgagttgt tgcccatact
     1861 taccaataaa tgccacatac ccgacctccg ggttaaatgt agctgcattc cgttgtggta
     1921 catgcggccg ctggcgggtg tgtcgagtgg atggtaggat cgacaaagat ctgatctgac
     1981 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     2041 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     2101 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     2161 atccacgccc attgatgtac tgccaaaacc gcatcagcat ggtaatagcg atgactaata
     2221 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     2281 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     2341 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     2401 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     2461 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     2521 tatgaactaa tgaccccgta attgattact agatctggct acactcgatc agcagttaga
     2581 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcaa
     2641 cgattgatag cataacccct tggggcctct aaacgggtct tgaggggttt tttgaattga
     2701 gaggtagctg aagctgctgt tatgaccact agagggaaga agatacctgt ggagctaatg
     2761 gtccaagatg tcgggagccg cgatcgctgc caaaaaggta agctgacgat agctgacgag
     2821 attcgatgcc cgacgagcaa atgagcgcca cggctgttcg cgtaagctcg aaacgacgga
     2881 gattcgatag agccagagtc gagtttaaac gaccttaagg ataaatttct ggtaaggagg
     2941 acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct ggcatatgtg
     3001 ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg cacctagaat
     3061 cggtcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggcgcggc
     3121 gataccgtaa agcacaagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     3181 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagacggc cacagtcgat
     3241 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccgtgggt
     3301 cacgacgaga tcctcgccgt cgggcataga cgccttgagc ctggcgaaca gttcggctgg
     3361 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     3421 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     3481 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     3541 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     3601 ttcagtgaca acgtcgagca cagccgcgca aggaacgccc gtcgtcgcca gccacgatag
     3661 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     3721 aaccggacgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     3781 ttgtgcccag tcatagccga atagcctctc cacccaagcc gccggagaac ctgcgtgcaa
     3841 tccatcttgt tcaatcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc
     3901 aaaagctagg actcgattat acgttgaccg atcagactat caatatctag gtttaaacag
     3961 atcggtacgc tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc
     4021 ccaagctgga ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac
     4081 tgagaggatc gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca
     4141 cacatccatc ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagcct
     4201 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     4261 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
     4321 ggaaggcggg caccccggcg cgctcggcgg cctccactcc ggggagcacg acggcgctgc
     4381 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
     4441 ccttgggccg gtgcggcgcc aggaggccct ccatctgttg ctgcgcggcc agccgggaac
     4501 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     4561 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     4621 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
     4681 ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
     4741 cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgt
     4801 tgtctaacaa aaaagccaaa aacggccaga atttagcgga caatttacaa gtctaacact
     4861 gaaaattaca tattgaccca aatgattaca tttcaaaagg tgcctaaaaa acttcacaaa
     4921 acacactcgc caaccccgag cgcatagttc aaaaccggag cttcagctac ttaagaagat
     4981 aggtacataa aaccgaccaa agaaactgac gcctcactta tccctcccct caccagaggt
     5041 ccggcgcctg tcgattcagg agagcctacc ctaggcccga accctgcgtc ctgcgacgga
     5101 gaaaagccta ccgcacacct accggcaggt ggccccaccc tgcattataa gccaacagaa
     5161 cgggtgacgt cacgacacga cgagggcgcg cgctcccaaa ggtacgggtg cactgcccaa
     5221 cggcaccgcc ataactgccg cccccgcaac agacgacaaa ccgagttctc cagtcagtga
     5281 caaacttcac gtcagggtcc ccagatggtg cccagcccat ctcacccgaa taagagcttt
     5341 cccgcattag cgaagggctc aagaccttgg gttcttgccg cccaccatgc cccccacctt
     5401 gtttcaacga cctcacagcc cgcctcacaa gcgtcttcca ttcaagactc gggaacagcc
     5461 gccattttgc tgcgctcccc ccaaccccca gttcagggca accttgctcg cggacccaga
     5521 ctacagccct tggcggtctc tccacacgct tccgtcccac cgagcggccc ggcggccacg
     5581 aaagccccgg ccagcccagc agcccgctac tcaccaagtg acgatcacag cgatccacaa
     5641 acaagaaccg cgacccaaat cccggctgcg acggaactag ctgtgccaca cccggcgcgt
     5701 ccttatataa tcatcggcgt tcaccgcccc acggagatcc ctccgcagaa tcgccgagaa
     5761 gggactactt ttcctcgcct gttccgctct ctggaaagaa aaccagtgcc ctagagtcac
     5821 ccaagtcccg tcctaaaatg tccttctgct gatactgggg ttctaaggcc gagtcttatg
     5881 agcagcgggc cgctgtcctg agcgtccggg cggaaggatc aggacgctcc tgcgcccttc
     5941 gtctgacgtg gcagcgctcg ccgtgacgag gggggcgccc gcgggaggcg ccaaaacccg
     6001 gcgcggaggc caggcgcgcc ccgatttcga tattcctcgg atctcggata gatcgggatt
     6061 gaggatgtag gttgaacttc tctcgggtga tcccgatctc ggatattcgg gattggagta
     6121 atacctggcg ataggcgctg ccaagcaata gccaaggcgc aaggccggcc aagagacttg
     6181 cgatagttcg ttcgagaggt agcctgcaag atggatcgga cgaaaggttt gaagattttc
     6241 cgacgagggc cttggagagt tcggaaattt aaatgcaact atggatgaac gaaatagaca
     6301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagttta
//
