LOCUS       sequence-337012-        8361 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 8361)
  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 8361)
  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..8361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5473..5496
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5500..5523
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5530..5553
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5593..5616
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5718..5737)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5783..5838
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5837..5856)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6197..6213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6197..6213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6210..6232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6221..6237
                     /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(6245..6251,6252..6269,6270..6275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6290..6311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6440..6457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6611..7199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6691..6710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7370..8161,8162..8230))
                     /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     7993..8012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8231..8335)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gcgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattgcg gccgtctcag
     1741 gccgagttcg gtaccgccac catgttcgtg ttcctggtgc tcctgcctct ggtgagcagc
     1801 cagtgcgtga acttcaccac ccgaacccag ctcccaccag cctacaccaa cagctttaca
     1861 cggggcgtgt actaccctga caaggtgttc agatctagcg tcctgcacag cactcaggac
     1921 ctcttcctgc cgttcttcag caacgtgaca tggttccacg ccatccacgt gagcggcaca
     1981 aacggaacca agcggtttgc taaccccgtc ctgccattca atgatggagt ttacttcgcc
     2041 agtaccgaga agagtaacat catccggggc tggatcttcg gcaccaccct ggatagcaaa
     2101 acacagagcc tcctgatcgt gaacaatgcc acgaacgtcg tgatcaaggt gtgcgagttc
     2161 cagttttgca atgatccttt cctgggtgtg tactaccaca agaacaacaa gagctggatg
     2221 gaaagcgagt tcagagtcta cagcagcgcc aacaactgca cattcgagta cgtctctcag
     2281 ccttttctga tggaccttga ggggaaacaa ggcaacttca agaacctgag agaattcgtg
     2341 ttcaagaaca tcgacggcta cttcaaaatc tactccaagc acacacccat caacctggtc
     2401 cggggcctcc ctcagggctt cagcgccctg gaacccctgg tcgacctgcc cataggcatc
     2461 aacataacgc ggttccaaac cctgcataga tcctacctga ctcctggcga cagcagcagc
     2521 ggatggaccg ccggagctgc agcctactat gtgggctacc tgcaacctag aaccttcctg
     2581 ctgaagtaca acgagaacgg cacaatcaca gacgccgtcg actgcgccct ggaccctctc
     2641 tctgagacaa agtgcaccct gaagtccttc accgtggaaa agggcatcta ccagaccagc
     2701 aacttccggg tgcagcctac agagagcatc gtgcgatttc caaacattac caacctctgc
     2761 cccttcggcg aggtgtttaa cgccacaaga tttgcctccg tttacgcctg gaatagaaag
     2821 agaatcagca attgtgtggc cgactactcc gtgctgtata acagcgcctc tttcagcacc
     2881 ttcaagtgct acggcgtttc cccaacaaag ctgaatgacc tgtgcttcac caacgtgtac
     2941 gccgactcct tcgtaattag aggcgatgag gtgcggcaga tcgcaccagg ccagaccggt
     3001 aacatcgctg actacaacta taagctgcct gatgatttta caggctgcgt gatcgcctgg
     3061 aactctaaca acctggatag caaggtgggc ggcaactaca actacctgta ccggctgttt
     3121 cgcaagtcta acctgaaacc tttcgagaga gacatctcca cagagatcta ccaggccggt
     3181 tctacacctt gtaacggggt gaaaggcttc aactgttact tccctctgca aagctacggc
     3241 ttccagccta cctacggagt cggctaccag ccataccggg tggtcgtgct gtccttcgag
     3301 ttactccacg cccccgccac cgtctgcggt cctaagaagt ccaccaatct ggttaagaac
     3361 aaatgcgtga acttcaactt caacggcctg accgggaccg gcgtgctgac cgaaagcaac
     3421 aaaaagttcc tccccttcca gcagttcggc cgtgatatcg ctgacaccac agatgccgtc
     3481 agagatccac agaccctgga aatcctggat attacaccct gctccttcgg aggagtttct
     3541 gtgatcaccc ccgggaccaa taccagcaac caggtggctg tgctgtacca aggtgttaac
     3601 tgcaccgagg ttcctgtggc catccacgcc gatcagctga cacctacttg gagagtgtac
     3661 tccactggct ccaatgtgtt ccagaccagg gccggatgtc tgatcggcgc cgagcacgtg
     3721 aataacagtt acgagtgcga catccctatc ggcgccggca tctgtgccag ctaccagacc
     3781 cagacaaaca gccctgggtc tgcttcctct gtagctagcc agagcatcat cgcctacacc
     3841 atgagcctgg gcgtggagaa cagcgtggcc tattccaaca actctatcgc cattcccacc
     3901 aactttacaa ttagcgtcac aacagagatc ctgcccgtga gcatgaccaa gaccagcgtg
     3961 gactgtacaa tgtacatctg tggcgacagc actgaatgca gcaacctgct gctgcaatac
     4021 ggctcctttt gcacccaact gaaccgggcg ctgaccggaa tcgccgtgga acaggacaaa
     4081 aatacccagg aggtgttcgc ccaagtgaag cagatctaca agaccccacc tatcaaggac
     4141 ttcggcggct ttaactttag ccagattctc cctgatcctt ctaagcctag caagcggagc
     4201 tttatcgagg atctgctgtt caacaaggtc accctggccg atgccggctt tatcaaacag
     4261 tatggcgatt gcctgggcga catagccgcc agagatctga tctgcgccca gaaattcaac
     4321 ggcctgacag ttctcccacc tctgctgacc gacgagatga tcgctcagta cacctctgcc
     4381 ctgctggctg gcaccatcac atctgggtgg acatttggcg ccggcgccgc cctgcagatc
     4441 ccctttgcca tgcagatggc ctatagattc aacggaatcg gcgtgaccca gaacgtgctg
     4501 tatgaaaacc agaagctgat cgctaaccag ttcaattctg ccatcggcaa gatccaggac
     4561 tccctctcct ctaccgccag cgccctgggc aaactgcagg acgtggtgaa tcagaacgcc
     4621 caagccctga acaccctggt gaagcagctc agcagcaatt ttggcgccat cagctctgtg
     4681 ctgaacgata tcctgtctag actggacaag gtggaagccg aagtccagat cgatagactg
     4741 atcacaggta gactgcagtc cctgcaaacc tacgtgaccc aacagctgat cagggccgct
     4801 gaaataagag ccagcgccaa tctcgccgct accaagatgt ccgagtgtgt gctgggacag
     4861 tctaaacgcg ttgacttctg cggcaaaggc tatcacctga tgagcttccc ccagagcgcg
     4921 ccgcacggcg tggtgttcct gcatgtgaca tacgtgcctg cccaagagaa gaatttcaca
     4981 accgcccctg ccatctgcca cgacggcaag gcccacttcc caagagaggg cgttttcgtt
     5041 tccaatggca cacactggtt cgtgacacaa agaaacttct acgaacccca gattatcacc
     5101 accgacaaca ccttcgtgag tggcaattgt gacgtggtca tcggaatcgt gaacaacaca
     5161 gtgtacgacc ctctgcaacc tgagctggac tcttttaagg aagagctgga caagtacttt
     5221 aaaaaccaca ccagccccga tgtggacctg ggcgacatca gtggcattaa cgccagcgtg
     5281 gtgaacatcc aaaaggaaat cgacagactg aacgaggtgg ccaagaacct gaacgagtcc
     5341 ctgatcgacc tgcaggagct cggcaaatac gagcagggat ccggatacat ccccgaggcc
     5401 cccagagatg gccaggccta cgtgcggaag gacggcgagt gggtactgct gagcacattc
     5461 ctgggcagat ccctggaggt gctgttccag ggcccaggcc atcaccacca tcaccaccat
     5521 catagcgcct ggtcccaccc ccagttcgag aagggcggcg gtagtggagg gggcggatct
     5581 ggcggctcag cttggagcca cccccagttc gaaaagtgat aatgactcga gcgataattc
     5641 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     5701 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     5761 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     5821 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     5881 aatgagtatt tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag
     5941 gtggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     6001 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     6061 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     6121 actactccca gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa
     6181 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6241 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6361 agcggatcga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6421 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6481 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6781 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7021 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7381 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7441 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7501 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7561 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7621 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7681 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7741 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7801 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7861 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7921 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7981 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8041 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8101 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8161 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8221 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8281 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8341 ttccccgaaa agtgccacct g
//
