LOCUS       sequence-308759-        5751 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 5751)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5751)
  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-m4133153
FEATURES             Location/Qualifiers
     source          1..5751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2080..2100
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2101..2145
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    2198..2319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2209..2228)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2263..2282
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2434..2480
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2510..2646
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2691..2762
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2988..3576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3068..3087)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4322..4390,4391..5182)
                     /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(4540..4559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5195..5331
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5483..5554
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5559..5605)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5623..5671
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgattgt taataattgg
      601 ttgaagcagt taattaaagt tacacttgtg ttcctttttg ttgctgctat tttctattta
      661 ataacacctg ttcatgtcat gtctaaacat actgactttt caagtgaaat cataggatac
      721 aaggctattg atggtggtgt cactcgtgac atagcatcta cagatacttg ttttgctaac
      781 aaacatgctg attttgacac atggtttagc cagcgtggtg gtagttatac taatgacaaa
      841 gcttgcccat tgattgctgc agtcataaca agagaagtgg gttttgtcgt gcctggtttg
      901 cctggcacga tattacgcac aactaatggt gactttttgc atttcttacc tagagttttt
      961 agtgcagttg gtaacatctg ttacacacca tcaaaactta tagagtacac tgactttgca
     1021 acatcagctt gtgttttggc tgctgaatgt acaattttta aagatgcttc tggtaagcca
     1081 gtaccatatt gttatgatac caatgtacta gaaggttctg ttgcttatga aagtttacgc
     1141 cctgacacac gttatgtgct catggatggc tctattattc aatttcctaa cacctacctt
     1201 gaaggttctg ttagagtggt aacaactttt gattctgagt actgtaggca cggcacttgt
     1261 gaaagatcag aagctggtgt ttgtgtatct actagtggta gatgggtact taacaatgat
     1321 tattacagat ctttaccagg agttttctgt ggtgtagatg ctgtaaattt acttactaat
     1381 atgtttacac cactaattca acctattggt gctttggaca tatcagcatc tatagtagct
     1441 ggtggtattg tagctatcgt agtaacatgc cttgcctact attttatgag gtttagaaga
     1501 gcttttggtg aatacagtca tgtagttgcc tttaatactt tactattcct tatgtcattc
     1561 actgtactct gtttaacacc agtttactca ttcttacctg gtgtttattc tgttatttac
     1621 ttgtacttga cattttatct tactaatgat gtttcttttt tagcacatat tcagtggatg
     1681 gttatgttca cacctttagt acctttctgg ataacaattg cttatatcat ttgtatttcc
     1741 acaaagcatt tctattggtt ctttagtaat tacctaaaga gacgtgtagt ctttaatggt
     1801 gtttccttta gtacttttga agaagctgcg ctgtgcacct ttttgttaaa taaagaaatg
     1861 tatctaaagt tgcgtagtga tgtgctatta cctcttacgc aatataatag atacttagct
     1921 ctttataata agtacaagta ttttagtgga gcaatggata caactagcta cagagaagct
     1981 gcttgttgtc atctcgcaaa ggctctcaat gacttcagta actcaggttc tgatgttctt
     2041 taccaaccac cacaaacctc tatcacctca gctgttttgg aaaaccttta ctttcagtcg
     2101 aaagaaaccg cggcagctaa gttcgagagg cagcacatgg acagttaagc tagcttgact
     2161 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     2221 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2281 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2341 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2401 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     2461 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     2521 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     2581 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     2641 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     2701 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     2761 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     2821 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     2881 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     2941 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3001 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3061 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3121 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3181 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3241 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3301 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3361 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3421 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3481 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3541 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3601 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3661 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3721 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3781 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     3841 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     3901 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     3961 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     4021 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     4081 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4141 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4201 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4261 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4321 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4381 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4441 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4501 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4561 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4621 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4681 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4741 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4801 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4861 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     4921 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4981 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5041 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5101 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5161 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5221 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5281 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     5341 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     5401 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     5461 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     5521 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     5581 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     5641 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     5701 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
