LOCUS       sequence-337764-        8380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein 
            S-GSAS-D614G-B.1.351 variant (South African).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8380)
  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 8380)
  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
FEATURES             Location/Qualifiers
     source          1..8380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          122..1138
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1062..1084
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             4946..4969
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4973..4996
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5003..5026
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5066..5089
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5191..5210)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5256..5311
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5310..5329)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5672..5688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5672..5688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5685..5707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5696..5712
                     /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(5720..5726,5727..5744,5745..5750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5765..5786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5915..5932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6086..6674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6166..6185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6845..7636,7637..7705))
                     /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     7468..7487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7706..7810)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7841..8220
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8222..8380,1..121)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
       61 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      121 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      181 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      241 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      301 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      361 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      421 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
      481 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
      541 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
      601 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
      661 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
      721 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
      781 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
      841 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
      901 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
      961 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1021 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1081 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1141 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1201 tcaggccgag ttcggtaccg ccaccatgtt cgtgttcctg gtgctcctgc ctctggtgag
     1261 cagccagtgc gtgaacttca ccacccgaac ccagctccca ccagcctaca ccaacagctt
     1321 tacacggggc gtgtactacc ctgacaaggt gttcagatct agcgtcctgc acagcactca
     1381 ggacctcttc ctgccgttct tcagcaacgt gacatggttc cacgccatcc acgtgagcgg
     1441 cacaaacgga accaagcggt ttgctaaccc cgtcctgcca ttcaatgatg gagtttactt
     1501 cgccagtacc gagaagagta acatcatccg gggctggatc ttcggcacca ccctggatag
     1561 caaaacacag agcctcctga tcgtgaacaa tgccacgaac gtcgtgatca aggtgtgcga
     1621 gttccagttt tgcaatgatc ctttcctggg tgtgtactac cacaagaaca acaagagctg
     1681 gatggaaagc gagttcagag tctacagcag cgccaacaac tgcacattcg agtacgtctc
     1741 tcagcctttt ctgatggacc ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt
     1801 cgtgttcaag aacatcgacg gctacttcaa aatctactcc aagcacacac ccatcaacct
     1861 ggtccggggc ctccctcagg gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg
     1921 catcaacata acgcggttcc aaaccctgct ggccctgcat atctcctacc tgactcctgg
     1981 cgacagcagc agcggatgga ccgccggagc tgcagcctac tatgtgggct acctgcaacc
     2041 tagaaccttc ctgctgaagt acaacgagaa cggcacaatc acagacgccg tcgactgcgc
     2101 cctggaccct ctctctgaga caaagtgcac cctgaagtcc ttcaccgtgg aaaagggcat
     2161 ctaccagacc agcaacttcc gggtgcagcc tacagagagc atcgtgcgat ttccaaacat
     2221 taccaacctc tgccccttcg gcgaggtgtt taacgccaca agatttgcct ccgtttacgc
     2281 ctggaataga aagagaatca gcaattgtgt ggccgactac tccgtgctgt ataacagcgc
     2341 ctctttcagc accttcaagt gctacggcgt ttccccaaca aagctgaatg acctgtgctt
     2401 caccaacgtg tacgccgact ccttcgtaat tagaggcgat gaggtgcggc agatcgcacc
     2461 aggccagacc ggtaacatcg ctgactacaa ctataagctg cctgatgatt ttacaggctg
     2521 cgtgatcgcc tggaactcta acaacctgga tagcaaggtg ggcggcaact acaactacct
     2581 gtaccggctg tttcgcaagt ctaacctgaa acctttcgag agagacatct ccacagagat
     2641 ctaccaggcc ggttctacac cttgtaacgg ggtgaaaggc ttcaactgtt acttccctct
     2701 gcaaagctac ggcttccagc ctacctacgg agtcggctac cagccatacc gggtggtcgt
     2761 gctgtccttc gagttactcc acgcccccgc caccgtctgc ggtcctaaga agtccaccaa
     2821 tctggttaag aacaaatgcg tgaacttcaa cttcaacggc ctgaccggga ccggcgtgct
     2881 gaccgaaagc aacaaaaagt tcctcccctt ccagcagttc ggccgtgata tcgctgacac
     2941 cacagatgcc gtcagagatc cacagaccct ggaaatcctg gatattacac cctgctcctt
     3001 cggaggagtt tctgtgatca cccccgggac caataccagc aaccaggtgg ctgtgctgta
     3061 ccaaggtgtt aactgcaccg aggttcctgt ggccatccac gccgatcagc tgacacctac
     3121 ttggagagtg tactccactg gctccaatgt gttccagacc agggccggat gtctgatcgg
     3181 cgccgagcac gtgaataaca gttacgagtg cgacatccct atcggcgccg gcatctgtgc
     3241 cagctaccag acccagacaa acagccctgg gtctgcttcc tctgtagcta gccagagcat
     3301 catcgcctac accatgagcc tgggcgtgga gaacagcgtg gcctattcca acaactctat
     3361 cgccattccc accaacttta caattagcgt cacaacagag atcctgcccg tgagcatgac
     3421 caagaccagc gtggactgta caatgtacat ctgtggcgac agcactgaat gcagcaacct
     3481 gctgctgcaa tacggctcct tttgcaccca actgaaccgg gcgctgaccg gaatcgccgt
     3541 ggaacaggac aaaaataccc aggaggtgtt cgcccaagtg aagcagatct acaagacccc
     3601 acctatcaag gacttcggcg gctttaactt tagccagatt ctccctgatc cttctaagcc
     3661 tagcaagcgg agctttatcg aggatctgct gttcaacaag gtcaccctgg ccgatgccgg
     3721 ctttatcaaa cagtatggcg attgcctggg cgacatagcc gccagagatc tgatctgcgc
     3781 ccagaaattc aacggcctga cagttctccc acctctgctg accgacgaga tgatcgctca
     3841 gtacacctct gccctgctgg ctggcaccat cacatctggg tggacatttg gcgccggcgc
     3901 cgccctgcag atcccctttg ccatgcagat ggcctataga ttcaacggaa tcggcgtgac
     3961 ccagaacgtg ctgtatgaaa accagaagct gatcgctaac cagttcaatt ctgccatcgg
     4021 caagatccag gactccctct cctctaccgc cagcgccctg ggcaaactgc aggacgtggt
     4081 gaatcagaac gcccaagccc tgaacaccct ggtgaagcag ctcagcagca attttggcgc
     4141 catcagctct gtgctgaacg atatcctgtc tagactggac aaggtggaag ccgaagtcca
     4201 gatcgataga ctgatcacag gtagactgca gtccctgcaa acctacgtga cccaacagct
     4261 gatcagggcc gctgaaataa gagccagcgc caatctcgcc gctaccaaga tgtccgagtg
     4321 tgtgctggga cagtctaaac gcgttgactt ctgcggcaaa ggctatcacc tgatgagctt
     4381 cccccagagc gcgccgcacg gcgtggtgtt cctgcatgtg acatacgtgc ctgcccaaga
     4441 gaagaatttc acaaccgccc ctgccatctg ccacgacggc aaggcccact tcccaagaga
     4501 gggcgttttc gtttccaatg gcacacactg gttcgtgaca caaagaaact tctacgaacc
     4561 ccagattatc accaccgaca acaccttcgt gagtggcaat tgtgacgtgg tcatcggaat
     4621 cgtgaacaac acagtgtacg accctctgca acctgagctg gactctttta aggaagagct
     4681 ggacaagtac tttaaaaacc acaccagccc cgatgtggac ctgggcgaca tcagtggcat
     4741 taacgccagc gtggtgaaca tccaaaagga aatcgacaga ctgaacgagg tggccaagaa
     4801 cctgaacgag tccctgatcg acctgcagga gctcggcaaa tacgagcagg gatccggata
     4861 catccccgag gcccccagag atggccaggc ctacgtgcgg aaggacggcg agtgggtact
     4921 gctgagcaca ttcctgggca gatccctgga ggtgctgttc cagggcccag gccatcacca
     4981 ccatcaccac catcatagcg cctggtccca cccccagttc gagaagggcg gcggtagtgg
     5041 agggggcgga tctggcggct cagcttggag ccacccccag ttcgaaaagt gataatgact
     5101 cgagcgataa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     5161 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     5221 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     5281 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     5341 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     5401 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5461 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5521 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     5581 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     5641 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5701 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5761 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5821 gaaacctgtc gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga
     5881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7801 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactagt
     7861 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7921 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7981 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     8041 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     8101 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     8161 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     8221 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     8281 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     8341 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
//
