LOCUS       sequence-363836-        8367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of SARS-CoV-2 Spike protein variant 
            OMICRON.BA.2 with furin side replaced by GSAS and with 2 Proline 
            substitution at 986 and 987.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8367)
  AUTHORS   Stalls V, Lindenberger J, Gobeil SM, Henderson R, Parks R, Barr M, 
            Deyton M, Martin M, Janowska K, Huang X, May A, Speakman M, Beaudoin
            E, Kraft B, Lu X, Edwards RJ, Eaton A, Montefiori DC, Williams WB, 
            Saunders KO, Wiehe K, Haynes BF, Acharya P
  TITLE     Cryo-EM structures of SARS-CoV-2 Omicron BA.2 spike.
  JOURNAL   Cell Rep. 2022 Jun 28;39(13):111009. doi: 
            10.1016/j.celrep.2022.111009. Epub 2022 Jun 8.
  PUBMED    35732171
REFERENCE   2  (bases 1 to 8367)
  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: "Cell Rep.";
            date: "2022-06-28"; volume: "39(13):111009. doi"; pages: " 
            10.1016/j.celrep.2022.111009. Epub 2022 Jun 8"
COMMENT     Sequence Label: p-alpha-H-S-GSAS-OMICRON.BA.2
FEATURES             Location/Qualifiers
     source          1..8367
                     /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             5477..5500
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5504..5527
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5534..5557
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5597..5620
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5722..5741)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5787..5842
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5841..5860)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6203..6219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6203..6219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6216..6238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6227..6243
                     /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(6251..6257,6258..6275,6276..6281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6296..6317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6446..6463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6617..7205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6697..6716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7376..8167,8168..8236))
                     /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     7999..8018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8237..8341)
                     /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 gggggcgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggcggagag gkgcggcggc 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 rgggctcggg 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 tcacccgaac ccagtcctac accaacagct ttacacgggg
     1861 cgtgtactac cctgacaagg tgttcagatc tagcgtcctg cacagcactc aggacctctt
     1921 cctgccgttc ttcagcaacg tgacatggtt ccacgccatc cacgtgagcg gcacaaacgg
     1981 aaccaagcgg tttgataacc ccgtcctgcc attcaatgat ggagtttact tcgccagtac
     2041 cgagaagagt aacatcatcc ggggctggat cttcggcacc accctggata gcaaaacaca
     2101 gagcctcctg atcgtgaaca atgccacgaa cgtcgtgatc aaggtgtgcg agttccagtt
     2161 ttgcaatgat cctttcctgg atgtgtacta ccacaagaac aacaagagct ggatggaaag
     2221 cgagttcaga gtctacagca gcgccaacaa ctgcacattc gagtacgtct ctcagccttt
     2281 tctgatggac cttgagggga aacaaggcaa cttcaagaac ctgagagaat tcgtgttcaa
     2341 gaacatcgac ggctacttca aaatctactc caagcacaca cccatcaacc tgggccggga
     2401 cctccctcag ggcttcagcg ccctggaacc cctggtcgac ctgcccatag gcatcaacat
     2461 aacgcggttc caaaccctgc tggccctgca tagatcctac ctgactcctg gcgacagcag
     2521 cagcggatgg accgccggag ctgcagccta ctatgtgggc tacctgcaac ctagaacctt
     2581 cctgctgaag tacaacgaga acggcacaat cacagacgcc gtcgactgcg ccctggaccc
     2641 tctctctgag acaaagtgca ccctgaagtc cttcaccgtg gaaaagggca tctaccagac
     2701 cagcaacttc cgggtgcagc ctacagagag catcgtgcga tttccaaaca ttaccaacct
     2761 ctgccccttc gacgaggtgt ttaacgccac aagatttgcc tccgtttacg cctggaatag
     2821 aaagagaatc agcaattgtg tggccgacta ctccgtgctg tataacttcg cccctttctt
     2881 cgccttcaag tgctacggcg tttccccaac aaagctgaat gacctgtgct tcaccaacgt
     2941 gtacgccgac tccttcgtaa ttagaggcaa tgaggtgtcc cagatcgcac caggccagac
     3001 cggtaacatc gctgactaca actataagct gcctgatgat tttacaggct gcgtgatcgc
     3061 ctggaactcc aacaagctgg atagcaaggt gggcggcaac tacaactacc tgtaccggct
     3121 gtttcgcaag tctaacctga aacctttcga gagagacatc tccacagaga tctaccaggc
     3181 cggtaacaag ccttgtaacg gggtggccgg cttcaactgt tacttccctc tgagaagcta
     3241 cggcttccgg cctacctatg gagtcggcca ccagccatac cgggtggtcg tgctgtcctt
     3301 cgagttactc cacgcccccg ccaccgtctg cggtcctaag aagtccacca atctggttaa
     3361 gaacaaatgc gtgaacttca acttcaacgg cctgaccggg accggcgtgc tgaccgaaag
     3421 caacaaaaag ttcctcccct tccagcagtt cggccgtgat atcgctgaca ccacagatgc
     3481 cgtcagagat ccacagaccc tggaaatcct ggatattaca ccctgctcct tcggaggagt
     3541 ttctgtgatc acccccggga ccaataccag caaccaggtg gctgtgctgt accaaggtgt
     3601 taactgcacc gaggttcctg tggccatcca cgccgatcag ctgacaccta cttggagagt
     3661 gtactccact ggctccaatg tgttccagac cagggccgga tgtctgatcg gcgccgagta
     3721 cgtgaataac agttacgagt gcgacatccc tatcggcgcc ggcatctgtg ccagctacca
     3781 gacccagaca aagagccatg ggtctgcttc ctctgtagct agccagagca tcatcgccta
     3841 caccatgagc ctgggcgcag agaacagcgt ggcctattcc aacaactcta tcgccattcc
     3901 caccaacttt acaattagcg tcacaacaga gatcctgccc gtgagcatga ccaagaccag
     3961 cgtggactgt acaatgtaca tctgtggcga cagcactgaa tgcagcaacc tgctgctgca
     4021 atacggctcc ttttgcaccc aactgaagcg ggcgctgacc ggaatcgccg tggaacagga
     4081 caaaaatacc caggaggtgt tcgcccaagt gaagcagatc tacaagaccc cacctatcaa
     4141 gtacttcggc ggctttaact ttagccagat tctccctgat ccttctaagc ctagcaagcg
     4201 gagctttatc gaggatctgc tgttcaacaa ggtcaccctg gccgatgccg gctttatcaa
     4261 acagtatggc gattgtctgg gcgacatagc cgccagagat ctgatctgcg cccagaaatt
     4321 caacggcctg acagttctcc cacctctgct gaccgacgag atgatcgctc agtacacctc
     4381 tgccctgctg gctggcacca tcacatctgg gtggacattt ggcgccggcg ccgccctgca
     4441 gatccccttt gccatgcaga tggcctatag attcaacgga atcggcgtga cccagaacgt
     4501 gctgtatgaa aaccagaagc tgatcgctaa ccagttcaat tctgccatcg gcaagatcca
     4561 ggactccctc tcctctaccg ccagcgccct gggcaaactg caggacgtgg tgaatcacaa
     4621 cgcccaagcc ctgaacaccc tggtgaagca gctcagcagc aagtttggcg ccatcagctc
     4681 tgtgctgaac gatatcctgt ctagactgga caaggtggaa gccgaagtcc agatcgatag
     4741 actgatcaca ggcagactgc agtccctgca aacctacgtg acccaacagc tgatcagggc
     4801 cgctgaaata agagccagcg ccaatctcgc cgctaccaag atgtccgagt gtgtgctggg
     4861 acagtctaaa cgcgttgact tctgcggcaa aggctatcac ctgatgagct tcccccagag
     4921 cgcgccgcac ggcgtggtgt tcctgcatgt gacatacgtg cctgcccaag agaagaattt
     4981 cacaaccgcc cctgccatct gccacgacgg caaggcccac ttcccaagag agggcgtttt
     5041 cgtttccaat ggcacacact ggttcgtgac acaaagaaac ttctacgaac cccagattat
     5101 caccaccgac aacaccttcg tgagtggcaa ttgtgacgtg gtcatcggaa tcgtgaacaa
     5161 cacagtgtac gaccctctgc aacctgagct ggactctttt aaggaagagc tggacaagta
     5221 ctttaaaaac cacaccagcc ccgatgtgga cctgggcgac atcagtggca ttaacgccag
     5281 cgtggtgaac atccaaaagg aaatcgacag actgaacgag gtggccaaga acctgaacga
     5341 gtccctgatc gacctgcagg agctcggcaa atacgagcag ggatccggat acatccccga
     5401 ggcccccaga gatggccagg cctacgtgcg gaaggacggc gagtgggtac tgctgagcac
     5461 attcctgggc agatccctgg aggtgctgtt ccagggccca ggccatcacc accatcacca
     5521 ccatcatagc gcctggtccc acccccagtt cgagaagggc ggcggtagtg gagggggcgg
     5581 atctggcggc tcagcttgga gccaccccca gttcgaaaag tgataatgac tcgagcgata
     5641 attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     5701 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     5761 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     5821 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     5881 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     5941 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     6001 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     6061 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     6121 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     6181 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6361 cgtgccagcg gatcgatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6421 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6481 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6541 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6601 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6661 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6721 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6781 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6841 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6901 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6961 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7021 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7081 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7141 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7201 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7261 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7321 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7381 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7441 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7501 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7561 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7621 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7681 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7741 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7801 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7861 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7921 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7981 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8041 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8101 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8161 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8221 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8281 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8341 gcacatttcc ccgaaaagtg ccacctg
//
