LOCUS       sequence-317254-        5733 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 5733)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5733)
  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-m4133660
FEATURES             Location/Qualifiers
     source          1..5733
                     /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..2127
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2180..2301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2191..2210)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2245..2264
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2416..2462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2492..2628
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2673..2744
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2970..3558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3050..3069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4304..4372,4373..5164)
                     /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(4522..4541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5177..5313
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5465..5536
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5541..5587)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5605..5653
                     /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 tacccctatg acgtaccaga ttatgcgtaa gctagcttga ctgactgaga tacagcgtac
     2161 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2221 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2281 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2341 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     2401 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2461 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     2521 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     2581 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     2641 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     2701 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     2761 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     2821 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     2881 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     2941 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3001 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3061 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3121 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3181 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3241 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3301 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3361 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3421 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3481 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3541 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3601 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3661 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3721 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3781 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     3841 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     3901 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     3961 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     4021 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc
     4081 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     4141 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     4201 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     4261 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     4321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     4441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     4501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     4621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4681 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     4741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4861 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     4921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5161 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     5221 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     5281 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     5341 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     5401 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     5461 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     5521 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     5581 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     5641 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     5701 gtcattagtt catagcccat atatggagtt ccg
//
