LOCUS       sequence-340773-        8368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein South African 
            variant version 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8368)
  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 8368)
  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.v2
FEATURES             Location/Qualifiers
     source          1..8368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5478..5501
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5505..5528
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5535..5558
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5598..5621
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5723..5742)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5788..5843
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5842..5861)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6204..6220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6204..6220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6217..6239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6228..6244
                     /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(6252..6258,6259..6276,6277..6282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6297..6318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6447..6464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6618..7206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6698..6717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7377..8168,8169..8237))
                     /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     8000..8019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8238..8342)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs grggcggrga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggrggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagcgcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga gttcggtacc gccaccatgt tcgtgttcct ggtgctcctg cctctggtga
     1801 gcagccagtg cgtgaacttc accacccgaa cccagctccc accagcctac accaacagct
     1861 ttacacgggg cgtgtactac cctgacaagg tgttcagatc tagcgtcctg cacagcactc
     1921 aggacctctt cctgccgttc ttcagcaacg tgacatggtt ccacgccatc cacgtgagcg
     1981 gcacaaacgg aaccaagcgg tttgctaacc ccgtcctgcc attcaatgat ggagtttact
     2041 tcgccagtac cgagaagagt aacatcatcc ggggctggat cttcggcacc accctggata
     2101 gcaaaacaca gagcctcctg atcgtgaaca atgccacgaa cgtcgtgatc aaggtgtgcg
     2161 agttccagtt ttgcaatgat cctttcctgg gtgtgtacta ccacaagaac aacaagagct
     2221 ggatggaaag cgagttcaga gtctacagca gcgccaacaa ctgcacattc gagtacgtct
     2281 ctcagccttt tctgatggac cttgagggga aacaaggcaa cttcaagaac ctgagagaat
     2341 tcgtgttcaa gaacatcgac ggctacttca aaatctactc caagcacaca cccatcaacc
     2401 tggtccgggg cctccctcag ggcttcagcg ccctggaacc cctggtcgac ctgcccatag
     2461 gcatcaacat aacgcggttc caaaccctgc atatctccta cctgactcct ggcgacagca
     2521 gcagcggatg gaccgccgga gctgcagcct actatgtggg ctacctgcaa cctagaacct
     2581 tcctgctgaa gtacaacgag aacggcacaa tcacagacgc cgtcgactgc gccctggacc
     2641 ctctctctga gacaaagtgc accctgaagt ccttcaccgt ggaaaagggc atctaccaga
     2701 ccagcaactt ccgggtgcag cctacagaga gcatcgtgcg atttccaaac attaccaacc
     2761 tctgcccctt cggcgaggtg tttaacgcca caagatttgc ctccgtttac gcctggaata
     2821 gaaagagaat cagcaattgt gtggccgact actccgtgct gtataacagc gcctctttca
     2881 gcaccttcaa gtgctacggc gtttccccaa caaagctgaa tgacctgtgc ttcaccaacg
     2941 tgtacgccga ctccttcgta attagaggcg atgaggtgcg gcagatcgca ccaggccaga
     3001 ccggtaacat cgctgactac aactataagc tgcctgatga ttttacaggc tgcgtgatcg
     3061 cctggaactc taacaacctg gatagcaagg tgggcggcaa ctacaactac ctgtaccggc
     3121 tgtttcgcaa gtctaacctg aaacctttcg agagagacat ctccacagag atctaccagg
     3181 ccggttctac accttgtaac ggggtgaaag gcttcaactg ttacttccct ctgcaaagct
     3241 acggcttcca gcctacctac ggagtcggct accagccata ccgggtggtc gtgctgtcct
     3301 tcgagttact ccacgccccc gccaccgtct gcggtcctaa gaagtccacc aatctggtta
     3361 agaacaaatg cgtgaacttc aacttcaacg gcctgaccgg gaccggcgtg ctgaccgaaa
     3421 gcaacaaaaa gttcctcccc ttccagcagt tcggccgtga tatcgctgac accacagatg
     3481 ccgtcagaga tccacagacc ctggaaatcc tggatattac accctgctcc ttcggaggag
     3541 tttctgtgat cacccccggg accaatacca gcaaccaggt ggctgtgctg taccaaggtg
     3601 ttaactgcac cgaggttcct gtggccatcc acgccgatca gctgacacct acttggagag
     3661 tgtactccac tggctccaat gtgttccaga ccagggccgg atgtctgatc ggcgccgagc
     3721 acgtgaataa cagttacgag tgcgacatcc ctatcggcgc cggcatctgt gccagctacc
     3781 agacccagac aaacagccct gggtctgctt cctctgtagc tagccagagc atcatcgcct
     3841 acaccatgag cctgggcgtg gagaacagcg tggcctattc caacaactct atcgccattc
     3901 ccaccaactt tacaattagc gtcacaacag agatcctgcc cgtgagcatg accaagacca
     3961 gcgtggactg tacaatgtac atctgtggcg acagcactga atgcagcaac ctgctgctgc
     4021 aatacggctc cttttgcacc caactgaacc gggcgctgac cggaatcgcc gtggaacagg
     4081 acaaaaatac ccaggaggtg ttcgcccaag tgaagcagat ctacaagacc ccacctatca
     4141 aggacttcgg cggctttaac tttagccaga ttctccctga tccttctaag cctagcaagc
     4201 ggagctttat cgaggatctg ctgttcaaca aggtcaccct ggccgatgcc ggctttatca
     4261 aacagtatgg cgattgcctg ggcgacatag ccgccagaga tctgatctgc gcccagaaat
     4321 tcaacggcct gacagttctc ccacctctgc tgaccgacga gatgatcgct cagtacacct
     4381 ctgccctgct ggctggcacc atcacatctg ggtggacatt tggcgccggc gccgccctgc
     4441 agatcccctt tgccatgcag atggcctata gattcaacgg aatcggcgtg acccagaacg
     4501 tgctgtatga aaaccagaag ctgatcgcta accagttcaa ttctgccatc ggcaagatcc
     4561 aggactccct ctcctctacc gccagcgccc tgggcaaact gcaggacgtg gtgaatcaga
     4621 acgcccaagc cctgaacacc ctggtgaagc agctcagcag caattttggc gccatcagct
     4681 ctgtgctgaa cgatatcctg tctagactgg acaaggtgga agccgaagtc cagatcgata
     4741 gactgatcac aggtagactg cagtccctgc aaacctacgt gacccaacag ctgatcaggg
     4801 ccgctgaaat aagagccagc gccaatctcg ccgctaccaa gatgtccgag tgtgtgctgg
     4861 gacagtctaa acgcgttgac ttctgcggca aaggctatca cctgatgagc ttcccccaga
     4921 gcgcgccgca cggcgtggtg ttcctgcatg tgacatacgt gcctgcccaa gagaagaatt
     4981 tcacaaccgc ccctgccatc tgccacgacg gcaaggccca cttcccaaga gagggcgttt
     5041 tcgtttccaa tggcacacac tggttcgtga cacaaagaaa cttctacgaa ccccagatta
     5101 tcaccaccga caacaccttc gtgagtggca attgtgacgt ggtcatcgga atcgtgaaca
     5161 acacagtgta cgaccctctg caacctgagc tggactcttt taaggaagag ctggacaagt
     5221 actttaaaaa ccacaccagc cccgatgtgg acctgggcga catcagtggc attaacgcca
     5281 gcgtggtgaa catccaaaag gaaatcgaca gactgaacga ggtggccaag aacctgaacg
     5341 agtccctgat cgacctgcag gagctcggca aatacgagca gggatccgga tacatccccg
     5401 aggcccccag agatggccag gcctacgtgc ggaaggacgg cgagtgggta ctgctgagca
     5461 cattcctggg cagatccctg gaggtgctgt tccagggccc aggccatcac caccatcacc
     5521 accatcatag cgcctggtcc cacccccagt tcgagaaggg cggcggtagt ggagggggcg
     5581 gatctggcgg ctcagcttgg agccaccccc agttcgaaaa gtgataatga ctcgagcgat
     5641 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     5701 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     5761 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     5821 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     5881 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     5941 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     6001 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     6061 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     6121 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     6181 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6241 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6301 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6361 tcgtgccagc ggatcgatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6421 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6481 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6541 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7021 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7441 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7501 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7561 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7621 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7681 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7741 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7801 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7861 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7921 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7981 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8041 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8101 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8161 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8221 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8341 cgcacatttc cccgaaaagt gccacctg
//
