LOCUS       sequence-300274-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5739)
  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"
COMMENT     Sequence Label: pGBW-m4134354
FEATURES             Location/Qualifiers
     source          1..5739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    32..103
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(108..154)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    172..220
                     /note="synthetic polyadenylation signal"
     enhancer        301..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2386..2406
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2407..2433
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2486..2607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2497..2516)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2551..2570
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2722..2768
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2798..2934
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2979..3050
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3356..3375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4610..4678,4679..5470)
                     /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     complement(4828..4847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5483..5619
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
       61 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
      121 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
      181 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
      241 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      841 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaagat tgtaaataac
      901 tggttgaaac aactaataaa ggttacttta gtgttcctgt ttgtcgccgc aatcttctat
      961 cttatcaccc cggtgcacgt gatgagtaaa catacagact tttcttcaga aattatcggg
     1021 tacaaggcta tcgatggcgg tgtaacgcgc gacattgcct ccacagatac ctgcttcgcg
     1081 aacaaacacg ctgactttga tacttggttc agccagcgag gaggctcgta caccaatgac
     1141 aaagcctgtc cactgatcgc agccgtgata actagagagg ttggattcgt cgtccctggg
     1201 ctccccggga caattctccg tacaaccaac ggcgactttc tgcattttct gccccgggtg
     1261 ttctccgcag tcggtaatat ctgttatact ccatctaagc tcattgagta cacggatttt
     1321 gctaccagcg cgtgcgtgtt ggccgcagaa tgcaccatat ttaaggacgc ttctggaaaa
     1381 cctgtgccct attgttacga taccaatgtg cttgagggct cagttgccta tgaaagtctg
     1441 aggccagaca ctagatacgt tttaatggat ggaagcatca tccagttccc aaacacgtat
     1501 ctggaggggt ccgtgcgggt cgtgaccaca ttcgattcag aatactgccg ccacggtaca
     1561 tgcgagaggt ctgaggctgg cgtatgtgtg tcaactagcg gtcgatgggt tctaaataac
     1621 gattattatc gttctctgcc cggcgttttt tgtggagtgg acgccgttaa ccttttgacc
     1681 aacatgttta cacctctcat tcagcccatt ggtgcactgg atatctccgc gagtattgtg
     1741 gctgggggaa tagtcgccat tgtcgtaacc tgcttggcat actatttcat gcgtttcagg
     1801 cgagcttttg gcgaatactc acatgtcgtg gccttcaata ctcttctgtt tctgatgagc
     1861 ttcacggtac tctgtctgac cccagtatac agtttcctac ctggggtgta ttctgtgatc
     1921 tacttatacc tcacatttta cctgaccaat gacgtgtcct ttctggccca catccaatgg
     1981 atggttatgt tcacgccgct cgtgcctttc tggattacaa tcgcttatat aatctgtatt
     2041 agcacgaagc atttctattg gttcttttcc aactacttaa agcggcgcgt cgtatttaat
     2101 ggggtatcgt ttagcacttt tgaggaagcc gcgctctgca ctttcttgct taataaggag
     2161 atgtatttga aattaagatc tgacgttctg cttcctttaa cacagtacaa cagatacctg
     2221 gcgttgtata acaaatacaa atatttctcg ggagccatgg atacaacttc ctatagagag
     2281 gctgcttgtt gccaccttgc aaaggcactc aacgacttca gtaacagcgg cagtgatgtg
     2341 ctttatcagc cccctcagac ttcaataaca tccgccgtcc tccaagaaaa cctttacttt
     2401 cagtcgtacc cctatgacgt accagattat gcgtaagcta gcttgactga ctgagataca
     2461 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     2521 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2581 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2641 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     2701 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     2761 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     2821 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     2881 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     2941 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     3001 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     3061 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     3121 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     3181 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     3241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4081 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     4141 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     4201 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     4261 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     4321 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat
     4381 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4441 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4501 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4561 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4621 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4681 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4741 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4801 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4861 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4921 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4981 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     5041 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5101 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5161 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5221 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5281 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5341 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5401 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5461 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5521 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5581 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5641 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5701 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagc
//
