LOCUS       sequence-352110-        8344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein South African 
            variant version 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8344)
  AUTHORS   Gobeil SM, Janowska K, McDowell S, Mansouri K, Parks R, Stalls V, 
            Kopp MF, Manne K, Li D, Wiehe K, Saunders KO, Edwards RJ, Korber B, 
            Haynes BF, Henderson R, Acharya P
  TITLE     Effect of natural mutations of SARS-CoV-2 on spike structure, 
            conformation, and antigenicity.
  JOURNAL   Science. 2021 Jun 24. pii: science.abi6226. doi: 
            10.1126/science.abi6226.
  PUBMED    34168071
REFERENCE   2  (bases 1 to 8344)
  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: "Science. 
            2021 Jun 24. pii: science.abi6226. doi: 10.1126/science.abi6226."
COMMENT     Sequence Label: p-alpha-H-S-GSAS-B.1.351.v4
FEATURES             Location/Qualifiers
     source          1..8344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          139..1155
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1079..1101
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             4954..4977
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4981..5004
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5011..5034
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5074..5097
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5199..5218)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5264..5319
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5318..5337)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5680..5696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5680..5696)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5693..5715)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5704..5720
                     /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(5728..5734,5735..5752,5753..5758))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5773..5794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5923..5940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6094..6682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6174..6193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6853..7644,7645..7713))
                     /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     7476..7495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7714..7818)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7849..8228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
       61 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      121 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      181 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      241 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      301 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      361 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
      421 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
      481 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
      541 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
      601 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
      661 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
      721 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
      781 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
      841 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
      901 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
      961 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     1021 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1081 ttcggcttct ggcgtgtgac cggcggctct agcgcctctg ctaaccatgt tcatgccttc
     1141 ttctttttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc
     1201 aaagaattgc ggccgtctca ggccgagttc ggtaccgcca ccatgttcgt gttcctggtg
     1261 ctcctgcctc tggtgagcag ccagtgcgtg aacttcacca cccgaaccca gctcccacca
     1321 gcctacacca acagctttac acggggcgtg tactaccctg acaaggtgtt cagatctagc
     1381 gtcctgcaca gcactcagga cctcttcctg ccgttcttca gcaacgtgac atggttccac
     1441 gccatccacg tgagcggcac aaacggaacc aagcggtttg ctaaccccgt cctgccattc
     1501 aatgatggag tttacttcgc cagtaccgag aagagtaaca tcatccgggg ctggatcttc
     1561 ggcaccaccc tggatagcaa aacacagagc ctcctgatcg tgaacaatgc cacgaacgtc
     1621 gtgatcaagg tgtgcgagtt ccagttttgc aatgatcctt tcctgggtgt gtactaccac
     1681 aagaacaaca agagctggat ggaaagcgag ttcagagtct acagcagcgc caacaactgc
     1741 acattcgagt acgtctctca gccttttctg atggaccttg aggggaaaca aggcaacttc
     1801 aagaacctga gagaattcgt gttcaagaac atcgacggct acttcaaaat ctactccaag
     1861 cacacaccca tcaacctggt ccggggcctc cctcagggct tcagcgccct ggaacccctg
     1921 gtcgacctgc ccataggcat caacataacg cggttccaaa ccctgcatag atcctacctg
     1981 actcctggcg acagcagcag cggatggacc gccggagctg cagcctacta tgtgggctac
     2041 ctgcaaccta gaaccttcct gctgaagtac aacgagaacg gcacaatcac agacgccgtc
     2101 gactgcgccc tggaccctct ctctgagaca aagtgcaccc tgaagtcctt caccgtggaa
     2161 aagggcatct accagaccag caacttccgg gtgcagccta cagagagcat cgtgcgattt
     2221 ccaaacatta ccaacctctg ccccttcggc gaggtgttta acgccacaag atttgcctcc
     2281 gtttacgcct ggaatagaaa gagaatcagc aattgtgtgg ccgactactc cgtgctgtat
     2341 aacagcgcct ctttcagcac cttcaagtgc tacggcgttt ccccaacaaa gctgaatgac
     2401 ctgtgcttca ccaacgtgta cgccgactcc ttcgtaatta gaggcgatga ggtgcggcag
     2461 atcgcaccag gccagaccgg taacatcgct gactacaact ataagctgcc tgatgatttt
     2521 acaggctgcg tgatcgcctg gaactctaac aacctggata gcaaggtggg cggcaactac
     2581 aactacctgt accggctgtt tcgcaagtct aacctgaaac ctttcgagag agacatctcc
     2641 acagagatct accaggccgg ttctacacct tgtaacgggg tgaaaggctt caactgttac
     2701 ttccctctgc aaagctacgg cttccagcct acctacggag tcggctacca gccataccgg
     2761 gtggtcgtgc tgtccttcga gttactccac gcccccgcca ccgtctgcgg tcctaagaag
     2821 tccaccaatc tggttaagaa caaatgcgtg aacttcaact tcaacggcct gaccgggacc
     2881 ggcgtgctga ccgaaagcaa caaaaagttc ctccccttcc agcagttcgg ccgtgatatc
     2941 gctgacacca cagatgccgt cagagatcca cagaccctgg aaatcctgga tattacaccc
     3001 tgctccttcg gaggagtttc tgtgatcacc cccgggacca ataccagcaa ccaggtggct
     3061 gtgctgtacc aaggtgttaa ctgcaccgag gttcctgtgg ccatccacgc cgatcagctg
     3121 acacctactt ggagagtgta ctccactggc tccaatgtgt tccagaccag ggccggatgt
     3181 ctgatcggcg ccgagcacgt gaataacagt tacgagtgcg acatccctat cggcgccggc
     3241 atctgtgcca gctaccagac ccagacaaac agccctgggt ctgcttcctc tgtagctagc
     3301 cagagcatca tcgcctacac catgagcctg ggcgtggaga acagcgtggc ctattccaac
     3361 aactctatcg ccattcccac caactttaca attagcgtca caacagagat cctgcccgtg
     3421 agcatgacca agaccagcgt ggactgtaca atgtacatct gtggcgacag cactgaatgc
     3481 agcaacctgc tgctgcaata cggctccttt tgcacccaac tgaaccgggc gctgaccgga
     3541 atcgccgtgg aacaggacaa aaatacccag gaggtgttcg cccaagtgaa gcagatctac
     3601 aagaccccac ctatcaagga cttcggcggc tttaacttta gccagattct ccctgatcct
     3661 tctaagccta gcaagcggag ctttatcgag gatctgctgt tcaacaaggt caccctggcc
     3721 gatgccggct ttatcaaaca gtatggcgat tgcctgggcg acatagccgc cagagatctg
     3781 atctgcgccc agaaattcaa cggcctgaca gttctcccac ctctgctgac cgacgagatg
     3841 atcgctcagt acacctctgc cctgctggct ggcaccatca catctgggtg gacatttggc
     3901 gccggcgccg ccctgcagat cccctttgcc atgcagatgg cctatagatt caacggaatc
     3961 ggcgtgaccc agaacgtgct gtatgaaaac cagaagctga tcgctaacca gttcaattct
     4021 gccatcggca agatccagga ctccctctcc tctaccgcca gcgccctggg caaactgcag
     4081 gacgtggtga atcagaacgc ccaagccctg aacaccctgg tgaagcagct cagcagcaat
     4141 tttggcgcca tcagctctgt gctgaacgat atcctgtcta gactggacaa ggtggaagcc
     4201 gaagtccaga tcgatagact gatcacaggt agactgcagt ccctgcaaac ctacgtgacc
     4261 caacagctga tcagggccgc tgaaataaga gccagcgcca atctcgccgc taccaagatg
     4321 tccgagtgtg tgctgggaca gtctaaacgc gttgacttct gcggcaaagg ctatcacctg
     4381 atgagcttcc cccagagcgc gccgcacggc gtggtgttcc tgcatgtgac atacgtgcct
     4441 gcccaagaga agaatttcac aaccgcccct gccatctgcc acgacggcaa ggcccacttc
     4501 ccaagagagg gcgttttcgt ttccaatggc acacactggt tcgtgacaca aagaaacttc
     4561 tacgaacccc agattatcac caccgacaac accttcgtga gtggcaattg tgacgtggtc
     4621 atcggaatcg tgaacaacac agtgtacgac cctctgcaac ctgagctgga ctcttttaag
     4681 gaagagctgg acaagtactt taaaaaccac accagccccg atgtggacct gggcgacatc
     4741 agtggcatta acgccagcgt ggtgaacatc caaaaggaaa tcgacagact gaacgaggtg
     4801 gccaagaacc tgaacgagtc cctgatcgac ctgcaggagc tcggcaaata cgagcaggga
     4861 tccggataca tccccgaggc ccccagagat ggccaggcct acgtgcggaa ggacggcgag
     4921 tgggtactgc tgagcacatt cctgggcaga tccctggagg tgctgttcca gggcccaggc
     4981 catcaccacc atcaccacca tcatagcgcc tggtcccacc cccagttcga gaagggcggc
     5041 ggtagtggag ggggcggatc tggcggctca gcttggagcc acccccagtt cgaaaagtga
     5101 taatgactcg agcgataatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     5161 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     5221 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     5281 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     5341 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     5401 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     5461 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     5521 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     5581 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     5641 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5701 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5761 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5821 ccagtcggga aacctgtcgt gccagcggat cgatccgctg cattaatgaa tcggccaacg
     5881 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5941 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6001 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat
     7861 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     7921 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     7981 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     8041 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     8101 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     8161 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     8221 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     8281 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     8341 gggg
//
