LOCUS       sequence-337363-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein 
            S-GSAS-D614G-deltaH69/V70 variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  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 8370)
  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-deltaH69/V70
FEATURES             Location/Qualifiers
     source          1..8370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5480..5503
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5507..5530
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5537..5560
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5600..5623
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5725..5744)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5790..5845
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5844..5863)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6206..6222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6206..6222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6219..6241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6230..6246
                     /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(6254..6260,6261..6278,6279..6284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6299..6320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6449..6466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6620..7208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6700..6719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7379..8170,8171..8239))
                     /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     8002..8021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8240..8344)
                     /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 gggggsgcgc gccrggsggg ggggggsggg gsgrggggsg
      541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1741 tcaggccgag ttcggtaccg ccaccatgtt cgtgttcctg gtgctcctgc ctctggtgag
     1801 cagccagtgc gtgaacctga ccacccgaac ccagctccca ccagcctaca ccaacagctt
     1861 tacacggggc gtgtactacc ctgacaaggt gttcagatct agcgtcctgc acagcactca
     1921 ggacctcttc ctgccgttct tcagcaacgt gacatggttc cacgccatca gcggcacaaa
     1981 cggaaccaag cggtttgata accccgtcct gccattcaat gatggagttt acttcgccag
     2041 taccgagaag agtaacatca tccggggctg gatcttcggc accaccctgg atagcaaaac
     2101 acagagcctc ctgatcgtga acaatgccac gaacgtcgtg atcaaggtgt gcgagttcca
     2161 gttttgcaat gatcctttcc tgggtgtgta ctaccacaag aacaacaaga gctggatgga
     2221 aagcgagttc agagtctaca gcagcgccaa caactgcaca ttcgagtacg tctctcagcc
     2281 ttttctgatg gaccttgagg ggaaacaagg caacttcaag aacctgagag aattcgtgtt
     2341 caagaacatc gacggctact tcaaaatcta ctccaagcac acacccatca acctggtccg
     2401 ggacctccct cagggcttca gcgccctgga acccctggtc gacctgccca taggcatcaa
     2461 cataacgcgg ttccaaaccc tgctggccct gcatagatcc tacctgactc ctggcgacag
     2521 cagcagcgga tggaccgccg gagctgcagc ctactatgtg ggctacctgc aacctagaac
     2581 cttcctgctg aagtacaacg agaacggcac aatcacagac gccgtcgact gcgccctgga
     2641 ccctctctct gagacaaagt gcaccctgaa gtccttcacc gtggaaaagg gcatctacca
     2701 gaccagcaac ttccgggtgc agcctacaga gagcatcgtg cgatttccaa acattaccaa
     2761 cctctgcccc ttcggcgagg tgtttaacgc cacaagattt gcctccgttt acgcctggaa
     2821 tagaaagaga atcagcaatt gtgtggccga ctactccgtg ctgtataaca gcgcctcttt
     2881 cagcaccttc aagtgctacg gcgtttcccc aacaaagctg aatgacctgt gcttcaccaa
     2941 cgtgtacgcc gactccttcg taattagagg cgatgaggtg cggcagatcg caccaggcca
     3001 gaccggtaag atcgctgact acaactataa gctgcctgat gattttacag gctgcgtgat
     3061 cgcctggaac tctaacaacc tggatagcaa ggtgggcggc aactacaact acctgtaccg
     3121 gctgtttcgc aagtctaacc tgaaaccttt cgagagagac atctccacag agatctacca
     3181 ggccggttct acaccttgta acggggtgga aggcttcaac tgttacttcc ctctgcaaag
     3241 ctacggcttc cagcctacca atggagtcgg ctaccagcca taccgggtgg tcgtgctgtc
     3301 cttcgagtta ctccacgccc ccgccaccgt ctgcggtcct aagaagtcca ccaatctggt
     3361 taagaacaaa tgcgtgaact tcaacttcaa cggcctgacc gggaccggcg tgctgaccga
     3421 aagcaacaaa aagttcctcc ccttccagca gttcggccgt gatatcgctg acaccacaga
     3481 tgccgtcaga gatccacaga ccctggaaat cctggatatt acaccctgct ccttcggagg
     3541 agtttctgtg atcacccccg ggaccaatac cagcaaccag gtggctgtgc tgtaccaagg
     3601 tgttaactgc accgaggttc ctgtggccat ccacgccgat cagctgacac ctacttggag
     3661 agtgtactcc actggctcca atgtgttcca gaccagggcc ggatgtctga tcggcgccga
     3721 gcacgtgaat aacagttacg agtgcgacat ccctatcggc gccggcatct gtgccagcta
     3781 ccagacccag acaaacagcc ctgggtctgc ttcctctgta gctagccaga gcatcatcgc
     3841 ctacaccatg agcctgggcg cagagaacag cgtggcctat tccaacaact ctatcgccat
     3901 tcccaccaac tttacaatta gcgtcacaac agagatcctg cccgtgagca tgaccaagac
     3961 cagcgtggac tgtacaatgt acatctgtgg cgacagcact gaatgcagca acctgctgct
     4021 gcaatacggc tccttttgca cccaactgaa ccgggcgctg accggaatcg ccgtggaaca
     4081 ggacaaaaat acccaggagg tgttcgccca agtgaagcag atctacaaga ccccacctat
     4141 caaggacttc ggcggcttta actttagcca gattctccct gatccttcta agcctagcaa
     4201 gcggagcttt atcgaggatc tgctgttcaa caaggtcacc ctggccgatg ccggctttat
     4261 caaacagtat ggcgattgcc tgggcgacat agccgccaga gatctgatct gcgcccagaa
     4321 attcaacggc ctgacagttc tcccacctct gctgaccgac gagatgatcg ctcagtacac
     4381 ctctgccctg ctggctggca ccatcacatc tgggtggaca tttggcgccg gcgccgccct
     4441 gcagatcccc tttgccatgc agatggccta tagattcaac ggaatcggcg tgacccagaa
     4501 cgtgctgtat gaaaaccaga agctgatcgc taaccagttc aattctgcca tcggcaagat
     4561 ccaggactcc ctctcctcta ccgccagcgc cctgggcaaa ctgcaggacg tggtgaatca
     4621 gaacgcccaa gccctgaaca ccctggtgaa gcagctcagc agcaattttg gcgccatcag
     4681 ctctgtgctg aacgatatcc tgtctagact ggacaaggtg gaagccgaag tccagatcga
     4741 tagactgatc acaggcagac tgcagtccct gcaaacctac gtgacccaac agctgatcag
     4801 ggccgctgaa ataagagcca gcgccaatct cgccgctacc aagatgtccg agtgtgtgct
     4861 gggacagtct aaacgcgttg acttctgcgg caaaggctat cacctgatga gcttccccca
     4921 gagcgcgccg cacggcgtgg tgttcctgca tgtgacatac gtgcctgccc aagagaagaa
     4981 tttcacaacc gcccctgcca tctgccacga cggcaaggcc cacttcccaa gagagggcgt
     5041 tttcgtttcc aatggcacac actggttcgt gacacaaaga aacttctacg aaccccagat
     5101 tatcaccacc gacaacacct tcgtgagtgg caattgtgac gtggtcatcg gaatcgtgaa
     5161 caacacagtg tacgaccctc tgcaacctga gctggactct tttaaggaag agctggacaa
     5221 gtactttaaa aaccacacca gccccgatgt ggacctgggc gacatcagtg gcattaacgc
     5281 cagcgtggtg aacatccaaa aggaaatcga cagactgaac gaggtggcca agaacctgaa
     5341 cgagtccctg atcgacctgc aggagctcgg caaatacgag cagggatccg gatacatccc
     5401 cgaggccccc agagatggcc aggcctacgt gcggaaggac ggcgagtggg tactgctgag
     5461 cacattcctg ggcagatccc tggaggtgct gttccagggc ccaggccatc accaccatca
     5521 ccaccatcat agcgcctggt cccaccccca gttcgagaag ggcggcggta gtggaggggg
     5581 cggatctggc ggctcagctt ggagccaccc ccagttcgaa aagtgataat gactcgagcg
     5641 ataattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     5701 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     5761 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     5821 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     5881 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     5941 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     6001 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     6061 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     6121 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     6181 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6241 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6301 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6361 tgtcgtgcca gcggatcgat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6421 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6481 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6541 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6601 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6661 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6721 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6781 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6841 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6901 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6961 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7021 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7081 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7141 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7201 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7261 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7321 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7381 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7441 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7501 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7561 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7621 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7681 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7741 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7801 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7861 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7921 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7981 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8041 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8101 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8161 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8221 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8281 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8341 cgcgcacatt tccccgaaaa gtgccacctg
//
