LOCUS       sequence-419021-        6116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally HA-tagged Schmallenberg 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 6116)
  TITLE     Plasmids encoding N- and C-terminally HA tagged Oropouche and 
            Schmallenberg orthobunyavirus proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6116)
  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            "Unpublished"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_SBV_Gc
FEATURES             Location/Qualifiers
     source          1..6116
                     /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    3255..3389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3279..3298)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3333..3352
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3500..3516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3500..3516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3513..3535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3524..3540
                     /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(3548..3554,3555..3572,3573..3578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3593..3614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3731..3748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3902..4490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3982..4001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4661..5452,5453..5521))
                     /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     5284..5303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5522..5626)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5694..5712
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5750..5772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5872..5891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6085..6107
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6099..6116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6100..6116
                     /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 tcgagttcac tatcaattca
      901 gcaagtaaca tacaatggaa agcactggcc cgaagtttca ccttaggagt gtgcaatacg
      961 aatcctcata aacatatatg tagatgcttg gagtctatgc aaatgtgcac atcaaccaag
     1021 acagaccacg ctagggaaat gtcaatatat tatgatggtc atccagatcg ctttgagcat
     1081 gacatgaaaa taatattgaa tataatgaga tatatagtcc ctggattagg tcgagtcttg
     1141 cttgatcaaa tcaaacaaac aaaagactac caagctttac gccacataca aggtaagctt
     1201 tctcctaaat cgcagtcaaa tttacaactt aaaggatttc tggaatttgt tgattttatc
     1261 cttggtgcaa acgtgacaat agaaaaaacc cctcaaacat taactacatt atctttgata
     1321 aaaggagccc acagaaactt ggatcaaaaa gatccaggtc caacaccaat actggtatgc
     1381 aaatcaccac aaaaagtggt atgctactca ccacgtggtg tcacacaccc aggagattat
     1441 atatcatgca aatctaagat gtataagtgg ccatctttag gggtatacaa acataataga
     1501 gaccagcaac aagcctgcag cagtgacaca cattgcctag agatgtttga accagcagaa
     1561 agaacaataa ctacaaaaat atgcaaagta agtgatatga cttattcaga atcgccatat
     1621 agtactggaa taccatcatg caacgtgaag agatttggat catgtaatgt aaggggtcat
     1681 caatggcaaa ttgcagaatg ctcaaatggc ttattttact atgtttcagc taaagcccat
     1741 tcgaaaacta acgatataac actgtactgt ttatcagcaa attgcctgga cttgcgttat
     1801 gcattcagat ccagtagttg ttcagatata gtatgggata caagttatcg aaataaatta
     1861 acacctaaat ctattaatca tccagatatt gaaaactaca tagcagcgct tcagtcagat
     1921 attgcaaatg atttaactat gcactacttt aaaccattaa aaaaccttcc agcaataatt
     1981 cctcaataca aaacaatgac attgaatggg gacaaggtat caaatggtat tagaaatagt
     2041 tatatcgagt cgcacatccc tgcaattaat ggtttatcag cagggattaa tattgccatg
     2101 ccaaatggag aaagcctctt ttccattatt atctatgtca gaagagtaat aaataaagca
     2161 tcgtatcgat ttctatatga aacaggaccc acaattggaa taaatgccaa gcacgaagag
     2221 gtatgtaccg ggaagtgccc aagcccaata ccacatcaag atggttgggt cacattctca
     2281 aaggaaagat caagtaattg gggctgtgaa gaatggggtt gcttggcaat aaatgatggt
     2341 tgtttatatg ggtcatgtca agacataata aggcctgaat ataagatata caagaagtct
     2401 agtattgaac aaaaggatgt tgaagtttgt ataaccatgg cccatgaatc attctgcagt
     2461 accgttgatg ttctccaacc tttaattagc gacaggatac aattagatat ccaaacgatt
     2521 caaatggact ctatgccaaa tataattgca gtcaagaatg ggaaagttta tgttggagat
     2581 atcaatgact taggttcgac agcaaagaaa tgtggctcag tccaattata ttctgaaggg
     2641 atcattggat cgggaacccc aaaatttgat tatgtttgcc atgcattcaa tcgtaaagat
     2701 gtcatccttc gaagatgctt tgataactca tatcagtctt gtcttctctt ggaacaagat
     2761 aatacattaa ctattgcttc taccagtcat atggaagtgc ataaaaaagt ttcaagcgtg
     2821 ggtacaatca attataaaat tatgttaggg gattttgact acaatgcata ttcaacacaa
     2881 gcaacagtca caatagatga gatcaggtgt ggtggttgtt atggctgccc tgaaggaatg
     2941 gcttgcgcac tcaaattgag taccaatacc atcgggagtt gttcaataaa aagtaactgc
     3001 gatacataca ttaaaataat agcagtcgat ccgatgcaga gcgagtattc cattaagtta
     3061 aactgcccac tagcaacaga gacagtttca gtaagtgtgt gctcagcttc tgcttacaca
     3121 aaaccttcaa tatctaaaaa tcaaccaaaa attgttttga attccttaga tgaaacatct
     3181 tacatcgagc aacatgataa aaagtgttct acatggcttt gcagagttta taaagaaggg
     3241 taggcggccg cggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     3301 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3361 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3421 aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc tgcaggcatg
     3481 ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3541 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3601 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3661 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3721 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3781 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4081 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4321 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4441 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4501 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4561 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4621 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4681 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4741 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4801 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4861 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4921 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4981 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5041 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5101 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5161 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5221 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5281 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5341 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5401 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5461 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5521 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5581 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5641 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5701 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5761 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5821 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     5881 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     5941 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6001 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6061 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagt
//
