LOCUS       sequence-339984-        8345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein South African 
            variant version 6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  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 8345)
  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.v6
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          134..1150
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1074..1096
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             4958..4981
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4985..5008
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5015..5038
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5078..5101
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5203..5222)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5268..5323
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5322..5341)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5684..5700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5684..5700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5697..5719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5708..5724
                     /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(5732..5738,5739..5756,5757..5762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5777..5798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5927..5944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6098..6686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6178..6197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6857..7648,7649..7717))
                     /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     7480..7499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7718..7822)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7853..8232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
       61 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      121 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      181 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      241 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      301 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      361 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
      421 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
      481 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
      541 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
      601 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
      661 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
      721 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
      781 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
      841 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
      901 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
      961 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1021 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1081 cttctggcgt gtgaccggcg gctctagcgc ctctgctaac catgttcatg ccttcttctt
     1141 tttcctacag ctcctgggca acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga
     1201 attgcggccg tctcaggccg agttcggtac cgccaccatg ttcgtgttcc tggtgctcct
     1261 gcctctggtg agcagccagt gcgtgaactt caccacccga acccagctcc caccagccta
     1321 caccaacagc tttacacggg gcgtgtacta ccctgacaag gtgttcagat ctagcgtcct
     1381 gcacagcact caggacctct tcctgccgtt cttcagcaac gtgacatggt tccacgccat
     1441 ccacgtgagc ggcacaaacg gaaccaagcg gtttgctaac cccgtcctgc cattcaatga
     1501 tggagtttac ttcgccagta ccgagaagag taacatcatc cggggctgga tcttcggcac
     1561 caccctggat agcaaaacac agagcctcct gatcgtgaac aatgccacga acgtcgtgat
     1621 caaggtgtgc gagttccagt tttgcaatga tcctttcctg ggtgtgtact accacaagaa
     1681 caacaagagc tggatggaaa gcgagttcag agtctacagc agcgccaaca actgcacatt
     1741 cgagtacgtc tctcagcctt ttctgatgga ccttgagggg aaacaaggca acttcaagaa
     1801 cctgagagaa ttcgtgttca agaacatcga cggctacttc aaaatctact ccaagcacac
     1861 acccatcaac ctggtccggg gcctccctca gggcttcagc gccctggaac ccctggtcga
     1921 cctgcccata ggcatcaaca taacgcggtt ccaaaccctg cacgccctgc atagatccta
     1981 cctgactcct ggcgacagca gcagcggatg gaccgccgga gctgcagcct actatgtggg
     2041 ctacctgcaa cctagaacct tcctgctgaa gtacaacgag aacggcacaa tcacagacgc
     2101 cgtcgactgc gccctggacc ctctctctga gacaaagtgc accctgaagt ccttcaccgt
     2161 ggaaaagggc atctaccaga ccagcaactt ccgggtgcag cctacagaga gcatcgtgcg
     2221 atttccaaac attaccaacc tctgcccctt cggcgaggtg tttaacgcca caagatttgc
     2281 ctccgtttac gcctggaata gaaagagaat cagcaattgt gtggccgact actccgtgct
     2341 gtataacagc gcctctttca gcaccttcaa gtgctacggc gtttccccaa caaagctgaa
     2401 tgacctgtgc ttcaccaacg tgtacgccga ctccttcgta attagaggcg atgaggtgcg
     2461 gcagatcgca ccaggccaga ccggtaacat cgctgactac aactataagc tgcctgatga
     2521 ttttacaggc tgcgtgatcg cctggaactc taacaacctg gatagcaagg tgggcggcaa
     2581 ctacaactac ctgtaccggc tgtttcgcaa gtctaacctg aaacctttcg agagagacat
     2641 ctccacagag atctaccagg ccggttctac accttgtaac ggggtgaaag gcttcaactg
     2701 ttacttccct ctgcaaagct acggcttcca gcctacctac ggagtcggct accagccata
     2761 ccgggtggtc gtgctgtcct tcgagttact ccacgccccc gccaccgtct gcggtcctaa
     2821 gaagtccacc aatctggtta agaacaaatg cgtgaacttc aacttcaacg gcctgaccgg
     2881 gaccggcgtg ctgaccgaaa gcaacaaaaa gttcctcccc ttccagcagt tcggccgtga
     2941 tatcgctgac accacagatg ccgtcagaga tccacagacc ctggaaatcc tggatattac
     3001 accctgctcc ttcggaggag tttctgtgat cacccccggg accaatacca gcaaccaggt
     3061 ggctgtgctg taccaaggtg ttaactgcac cgaggttcct gtggccatcc acgccgatca
     3121 gctgacacct acttggagag tgtactccac tggctccaat gtgttccaga ccagggccgg
     3181 atgtctgatc ggcgccgagc acgtgaataa cagttacgag tgcgacatcc ctatcggcgc
     3241 cggcatctgt gccagctacc agacccagac aaacagccct gggtctgctt cctctgtagc
     3301 tagccagagc atcatcgcct acaccatgag cctgggcgtg gagaacagcg tggcctattc
     3361 caacaactct atcgccattc ccaccaactt tacaattagc gtcacaacag agatcctgcc
     3421 cgtgagcatg accaagacca gcgtggactg tacaatgtac atctgtggcg acagcactga
     3481 atgcagcaac ctgctgctgc aatacggctc cttttgcacc caactgaacc gggcgctgac
     3541 cggaatcgcc gtggaacagg acaaaaatac ccaggaggtg ttcgcccaag tgaagcagat
     3601 ctacaagacc ccacctatca aggacttcgg cggctttaac tttagccaga ttctccctga
     3661 tccttctaag cctagcaagc ggagctttat cgaggatctg ctgttcaaca aggtcaccct
     3721 ggccgatgcc ggctttatca aacagtatgg cgattgcctg ggcgacatag ccgccagaga
     3781 tctgatctgc gcccagaaat tcaacggcct gacagttctc ccacctctgc tgaccgacga
     3841 gatgatcgct cagtacacct ctgccctgct ggctggcacc atcacatctg ggtggacatt
     3901 tggcgccggc gccgccctgc agatcccctt tgccatgcag atggcctata gattcaacgg
     3961 aatcggcgtg acccagaacg tgctgtatga aaaccagaag ctgatcgcta accagttcaa
     4021 ttctgccatc ggcaagatcc aggactccct ctcctctacc gccagcgccc tgggcaaact
     4081 gcaggacgtg gtgaatcaga acgcccaagc cctgaacacc ctggtgaagc agctcagcag
     4141 caattttggc gccatcagct ctgtgctgaa cgatatcctg tctagactgg acaaggtgga
     4201 agccgaagtc cagatcgata gactgatcac aggtagactg cagtccctgc aaacctacgt
     4261 gacccaacag ctgatcaggg ccgctgaaat aagagccagc gccaatctcg ccgctaccaa
     4321 gatgtccgag tgtgtgctgg gacagtctaa acgcgttgac ttctgcggca aaggctatca
     4381 cctgatgagc ttcccccaga gcgcgccgca cggcgtggtg ttcctgcatg tgacatacgt
     4441 gcctgcccaa gagaagaatt tcacaaccgc ccctgccatc tgccacgacg gcaaggccca
     4501 cttcccaaga gagggcgttt tcgtttccaa tggcacacac tggttcgtga cacaaagaaa
     4561 cttctacgaa ccccagatta tcaccaccga caacaccttc gtgagtggca attgtgacgt
     4621 ggtcatcgga atcgtgaaca acacagtgta cgaccctctg caacctgagc tggactcttt
     4681 taaggaagag ctggacaagt actttaaaaa ccacaccagc cccgatgtgg acctgggcga
     4741 catcagtggc attaacgcca gcgtggtgaa catccaaaag gaaatcgaca gactgaacga
     4801 ggtggccaag aacctgaacg agtccctgat cgacctgcag gagctcggca aatacgagca
     4861 gggatccgga tacatccccg aggcccccag agatggccag gcctacgtgc ggaaggacgg
     4921 cgagtgggta ctgctgagca cattcctggg cagatccctg gaggtgctgt tccagggccc
     4981 aggccatcac caccatcacc accatcatag cgcctggtcc cacccccagt tcgagaaggg
     5041 cggcggtagt ggagggggcg gatctggcgg ctcagcttgg agccaccccc agttcgaaaa
     5101 gtgataatga ctcgagcgat aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     5161 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     5221 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5281 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5341 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5401 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5461 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5521 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     5581 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     5641 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     5701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5821 ctttccagtc gggaaacctg tcgtgccagc ggatcgatcc gctgcattaa tgaatcggcc
     5881 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5941 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6001 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6061 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6121 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6181 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6241 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6301 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6361 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6421 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6481 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6541 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6601 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6661 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6721 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6781 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6841 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6901 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6961 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7021 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7081 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7141 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7201 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7261 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7321 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7381 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7441 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7501 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7561 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7621 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7681 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7741 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7801 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga
     7861 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7921 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7981 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     8041 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     8101 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     8161 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     8221 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     8281 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     8341 ggggg
//
