LOCUS       sequence-297929-        8055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein 
            (Spike protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8055)
  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-m4133326
FEATURES             Location/Qualifiers
     source          1..8055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3824..3844
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3845..3871
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3924..4045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3935..3954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3989..4008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4160..4206
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4236..4372
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4417..4488
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4714..5302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4794..4813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6048..6116,6117..6908)
                     /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(6266..6285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6921..7057
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7209..7280
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7285..7331)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7349..7397
                     /note="synthetic polyadenylation signal"
     enhancer        7478..7781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7783..7985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7936..7956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc gtctttcttg tgttactccc cctggtttcg tcccagtgcg tgaacttgac
       61 tacgcgaacc caactgccgc cagcatatac aaatagtttc acccgtggag tatactaccc
      121 tgataaagtg tttagatcaa gcgtactaca ctctactcag gacctgtttc tccccttctt
      181 ctccaatgtc acatggtttc atgcgatcca cgttagcggc accaacggga caaagcgctt
      241 tgacaaccct gtgctgccat tcaatgatgg tgtgtatttt gcttctaccg agaagagcaa
      301 cataattcgg ggctggatct tcggaactac cctcgacagt aaaactcagt cattgctgat
      361 tgtgaataac gccacaaatg tcgttatcaa ggtctgtgaa tttcaattct gcaacgatcc
      421 attccttggg gtgtattacc ataaaaataa taagtcctgg atggagagcg aatttagggt
      481 atactcgtct gccaacaatt gtactttcga gtatgtgtct cagcccttct taatggatct
      541 ggaaggtaaa cagggcaact ttaagaacct ccgcgagttt gtattcaaaa atatagacgg
      601 atactttaag atttattcca agcacacacc aatcaatttg gtgcgggatc tcccgcaagg
      661 gttcagcgct ctggaacctc tagtggacct ccccatcggc attaacatca caagattcca
      721 gacgctgtta gccctgcata gatcatacct tactcccggt gacagtagtt ctgggtggac
      781 cgcaggcgcg gctgcatatt acgtgggata tttgcagcct cggacgtttc tgctgaaata
      841 caacgagaac ggcaccatta cagatgccgt ggattgcgcc cttgaccccc taagcgaaac
      901 taagtgtacg cttaaaagct tcaccgtcga gaagggaatt taccagactt ccaactttag
      961 ggtccagccg acagaatcaa tagttaggtt cccaaatatc accaatttgt gcccttttgg
     1021 ggaagtcttc aatgctacac gatttgcttc ggtttatgcc tggaacagga agcgtatctc
     1081 aaattgcgtc gcggactaca gcgtgctcta taatagtgca agtttctcga cctttaaatg
     1141 ttatggcgtg tccccgacca aactaaatga cctctgtttt acaaacgttt acgccgattc
     1201 ttttgtgatt cgcggtgatg aggtacgcca aatcgcaccg ggccagactg gcaagattgc
     1261 cgattacaac tacaagttgc ctgacgactt caccggatgt gttatagcct ggaatagcaa
     1321 taatctggat tctaaggtcg gaggtaacta taactacctg tatagactgt tccgtaagtc
     1381 aaacctgaaa ccctttgagc gggacatctc cacagagatt taccaagcag ggtctacccc
     1441 ctgcaatggg gttgaaggat tcaattgcta ctttccatta cagagctacg ggttccagcc
     1501 tacgaacggg gttggttacc agccttatag ggtagtggtg ctgtcgttcg aactgttgca
     1561 cgctccagct acagtgtgcg ggcctaaaaa atcaactaac ctggttaaaa ataagtgtgt
     1621 taattttaac ttcaatggct taaccggaac tggagtgctc acggagtcaa acaagaaatt
     1681 tctaccgttt cagcaattcg ggagggatat cgcagacact acggacgcgg tcagagatcc
     1741 acagaccctt gagatccttg acataacacc ctgctctttc ggaggtgtct ccgtaattac
     1801 accaggcact aatacctcta atcaggtggc cgtgctttat caggatgtta actgtacaga
     1861 ggtcccggtg gctatccatg ccgaccaatt gacacctacg tggcgcgtct attctaccgg
     1921 ctccaacgtc ttccagaccc gggctggatg cttgataggc gcagaacacg taaacaactc
     1981 ttacgaatgt gatattccca ttggagccgg gatctgcgca agctaccaga cccaaacaaa
     2041 cagtcccaga cgggctcgta gcgtggcctc tcagagcata atagcctata ccatgtccct
     2101 tggcgcggag aatagtgtgg catactcaaa taatagcatc gctatcccca ctaactttac
     2161 tatttccgtg acgactgaga ttctgccagt gtctatgact aagacatccg tggattgcac
     2221 aatgtatatt tgtggggatt ccacagagtg tagcaacttg ttgctgcaat atggatcatt
     2281 ctgtacgcag ctgaatcgcg ccctcaccgg catcgctgtg gaacaggaca agaacactca
     2341 agaggtcttt gctcaggtga agcaaatcta caagacccca cctataaaag acttcggggg
     2401 ttttaacttc tcgcagattc tccccgaccc cagcaagcct tcaaaaagga gctttattga
     2461 ggacctgttg ttcaacaagg ttactttagc cgatgccggt tttatcaaac agtacggcga
     2521 ctgtcttggc gatatagcag cccgggatct gatttgcgca caaaaattca atggccttac
     2581 cgtactgcca ccccttctca ccgacgaaat gatcgcgcag tacacctccg cactgctggc
     2641 tgggaccatc acttcgggat ggacctttgg ggctggtgct gctttgcaaa ttcccttcgc
     2701 catgcagatg gcctatcgat tcaatggcat aggggtaaca cagaacgtat tgtatgaaaa
     2761 tcaaaagcta atcgctaacc agtttaattc agccattggt aagatccagg attcactctc
     2821 cagtacagcc tctgccctgg gaaagctcca ggacgtggtc aaccagaacg cgcaggcatt
     2881 gaatacactc gtcaaacagc tgagtagcaa tttcggcgca atcagtagtg tcttaaacga
     2941 tatactgagt aggcttgata aagtagaggc cgaggtccag attgacagac taatcacggg
     3001 gcgtcttcag agtttacaga cctatgtgac ccaacagctg atccgagcag cggaaattcg
     3061 agcaagcgcg aacctggccg ctactaaaat gagtgagtgc gtgctgggcc aaagtaaacg
     3121 agtcgacttt tgcggcaaag ggtatcatct catgtctttc cctcagtccg cccctcacgg
     3181 tgttgttttc ctccacgtga catacgttcc cgcgcaggaa aagaatttta ctacggcacc
     3241 cgctatttgc catgacggaa aggctcattt tccacgggag ggagtgttcg tttcaaacgg
     3301 gacacactgg tttgtaactc agcggaactt ttatgagcct caaatcatca cgacagacaa
     3361 taccttcgtg agtggcaact gcgatgtagt cataggcatt gtgaataaca ctgtttatga
     3421 tccacttcag ccggaactcg acagcttcaa agaggaattg gataagtact ttaaaaatca
     3481 taccagtcca gacgtagatc tcggcgatat atcgggcatt aacgccagtg tggtcaatat
     3541 acaaaaggag attgacagac tgaatgaagt agctaagaac ctcaacgaat ctctgattga
     3601 cctgcaagag ctagggaagt acgagcagta tatcaaatgg ccatggtaca tatggctggg
     3661 ttttatcgcg ggtcttattg ccatagtcat ggtgacgatt atgttgtgct gtatgacctc
     3721 ctgctgttct tgtttgaaag gctgctgctc ctgtggtagc tgttgcaaat tcgatgaaga
     3781 tgactctgaa cccgtgttga aaggcgttaa gctacactac acagaaaacc tttactttca
     3841 gtcgtacccc tatgacgtac cagattatgc gtaagctagc ttgactgact gagatacagc
     3901 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4021 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4081 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     4141 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     4201 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     4261 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     4321 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     4381 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     4441 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     4501 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     4561 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     4621 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     4681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5521 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     5581 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     5641 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     5701 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     5761 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     5821 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     5881 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     5941 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     6001 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     6061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     6181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     6241 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     6361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6421 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     6481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6601 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     6661 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6901 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     6961 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     7021 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     7081 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     7141 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     7201 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     7261 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     7321 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     7381 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     7441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     7621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     7681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     7741 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     7801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     7861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     7921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     7981 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     8041 cccgccagcg gccgc
//
