LOCUS       sequence-352869-        8369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of SARS-CoV-2 Spike protein variant 
            OMICRON with furin side intact.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8369)
  AUTHORS   Gobeil SM, Henderson R, Stalls V, Janowska K, Huang X, May A, 
            Speakman M, Beaudoin E, Manne K, Li D, Parks R, Barr M, Deyton M, 
            Martin M, Mansouri K, Edwards RJ, Eaton A, Montefiori DC, Sempowski 
            GD, Saunders KO, Wiehe K, Williams W, Korber B, Haynes BF, Acharya P
  TITLE     Structural diversity of the SARS-CoV-2 Omicron spike.
  JOURNAL   Mol Cell. 2022 Jun 2;82(11):2050-2068.e6. doi: 
            10.1016/j.molcel.2022.03.028. Epub 2022 Mar 25.
  PUBMED    35447081
REFERENCE   2  (bases 1 to 8369)
  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"; doi: 
            "10.1016/j.molcel.2022.03.028"; journalName: "Mol Cell"; date: 
            "2022-06-2- 2"; volume: "82"; issue: "11"; pages: "2050-2068.e6"
COMMENT     Sequence Label: p-alpha-H-S-RRAR-OMICRON
FEATURES             Location/Qualifiers
     source          1..8369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5475..5498
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5502..5525
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5532..5555
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5595..5618
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5720..5739)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5785..5840
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5839..5858)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6201..6217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6201..6217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6214..6236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6225..6241
                     /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(6249..6255,6256..6273,6274..6279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6294..6315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6444..6461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6615..7203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6695..6714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7374..8165,8166..8234))
                     /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     7997..8016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8235..8339)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc crggsggggs ggggsggggs grggggsggg
      541 gsggggsgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagcgcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttgtt gtgctgtctc atcattttgg caaagaattg cggccgtctc
     1741 aggccgagtt cggtaccgcc accatgttcg tgttcctggt gctcctgcct ctggtgagca
     1801 gccagtgcgt gaacctgacc acccgaaccc agctcccacc agcctacacc aacagcttta
     1861 cacggggcgt gtactaccct gacaaggtgt tcagatctag cgtcctgcac agcactcagg
     1921 acctcttcct gccgttcttc agcaacgtga catggttcca cgtcatcagc ggcacaaacg
     1981 gaaccaagcg gtttgataac cccgtcctgc cattcaatga tggagtttac ttcgccagta
     2041 tcgagaagag taacatcatc cggggctgga tcttcggcac caccctggat agcaaaacac
     2101 agagcctcct gatcgtgaac aatgccacga acgtcgtgat caaggtgtgc gagttccagt
     2161 tttgcaatga tcctttcctg gaccacaaga acaacaagag ctggatggaa agcgagttca
     2221 gagtctacag cagcgccaac aactgcacat tcgagtacgt ctctcagcct tttctgatgg
     2281 accttgaggg gaaacaaggc aacttcaaga acctgagaga attcgtgttc aagaacatcg
     2341 acggctactt caaaatctac tccaagcaca cacccatcat cgtccgggag cctgaagacc
     2401 tccctcaggg cttcagcgcc ctggaacccc tggtcgacct gcccataggc atcaacataa
     2461 cgcggttcca aaccctgctg gccctgcata gatcctacct gactcctggc gacagcagca
     2521 gcggatggac cgccggagct gcagcctact atgtgggcta cctgcaacct agaaccttcc
     2581 tgctgaagta caacgagaac ggcacaatca cagacgccgt cgactgcgcc ctggaccctc
     2641 tctctgagac aaagtgcacc ctgaagtcct tcaccgtgga aaagggcatc taccagacca
     2701 gcaacttccg ggtgcagcct acagagagca tcgtgcgatt tccaaacatt accaacctct
     2761 gccccttcga cgaggtgttt aacgccacaa gatttgcctc cgtttacgcc tggaatagaa
     2821 agagaatcag caattgtgtg gccgactact ccgtgctgta taacctcgcc cctttcttca
     2881 ccttcaagtg ctacggcgtt tccccaacaa agctgaatga cctgtgcttc accaacgtgt
     2941 acgccgactc cttcgtaatt agaggcgatg aggtgcggca gatcgcacca ggccagaccg
     3001 gtaacatcgc tgactacaac tataagctgc ctgatgattt tacaggctgc gtgatcgcct
     3061 ggaactctaa caagctggat agcaaggtga gcggcaacta caactacctg taccggctgt
     3121 ttcgcaagtc taacctgaaa cctttcgaga gagacatctc cacagagatc taccaggccg
     3181 gtaacaagcc ttgtaacggg gtggccggct tcaactgtta cttccctctg agaagctaca
     3241 gcttccggcc tacctatgga gtcggccacc agccataccg ggtggtcgtg ctgtccttcg
     3301 agttactcca cgcccccgcc accgtctgcg gtcctaagaa gtccaccaat ctggttaaga
     3361 acaaatgcgt gaacttcaac ttcaacggcc tgaaggggac cggcgtgctg accgaaagca
     3421 acaaaaagtt cctccccttc cagcagttcg gccgtgatat cgctgacacc acagatgccg
     3481 tcagagatcc acagaccctg gaaatcctgg atattacacc ctgctccttc ggaggagttt
     3541 ctgtgatcac ccccgggacc aataccagca accaggtggc tgtgctgtac caaggtgtta
     3601 actgcaccga ggttcctgtg gccatccacg ccgatcagct gacacctact tggagagtgt
     3661 actccactgg ctccaatgtg ttccagacca gggccggatg tctgatcggc gccgagtacg
     3721 tgaataacag ttacgagtgc gacatcccta tcggcgccgg catctgtgcc agctaccaga
     3781 cccagacaaa gagccataga cgggctagat ctgtagctag ccagagcatc atcgcctaca
     3841 ccatgagcct gggcgcagag aacagcgtgg cctattccaa caactctatc gccattccca
     3901 ccaactttac aattagcgtc acaacagaga tcctgcccgt gagcatgacc aagaccagcg
     3961 tggactgtac aatgtacatc tgtggcgaca gcactgaatg cagcaacctg ctgctgcaat
     4021 acggctcctt ttgcacccaa ctgaagcggg cgctgaccgg aatcgccgtg gaacaggaca
     4081 aaaataccca ggaggtgttc gcccaagtga agcagatcta caagacccca cctatcaagt
     4141 acttcggcgg ctttaacttt agccagattc tccctgatcc ttctaagcct agcaagcgga
     4201 gctttatcga ggatctgctg ttcaacaagg tcaccctggc cgatgccggc tttatcaaac
     4261 agtatggcga ttgcctgggc gacatagccg ccagagatct gatctgcgcc cagaaattca
     4321 agggcctgac agttctccca cctctgctga ccgacgagat gatcgctcag tacacctctg
     4381 ccctgctggc tggcaccatc acatctgggt ggacatttgg cgccggcgcc gccctgcaga
     4441 tcccctttgc catgcagatg gcctatagat tcaacggaat cggcgtgacc cagaacgtgc
     4501 tgtatgaaaa ccagaagctg atcgctaacc agttcaattc tgccatcggc aagatccagg
     4561 actccctctc ctctaccgcc agcgccctgg gcaaactgca ggacgtggtg aatcacaacg
     4621 cccaagccct gaacaccctg gtgaagcagc tcagcagcaa gtttggcgcc atcagctctg
     4681 tgctgaacga tatcttctct agactggaca aggtggaagc cgaagtccag atcgatagac
     4741 tgatcacagg cagactgcag tccctgcaaa cctacgtgac ccaacagctg atcagggccg
     4801 ctgaaataag agccagcgcc aatctcgccg ctaccaagat gtccgagtgt gtgctgggac
     4861 agtctaaacg cgttgacttc tgcggcaaag gctatcacct gatgagcttc ccccagagcg
     4921 cgccgcacgg cgtggtgttc ctgcatgtga catacgtgcc tgcccaagag aagaatttca
     4981 caaccgcccc tgccatctgc cacgacggca aggcccactt cccaagagag ggcgttttcg
     5041 tttccaatgg cacacactgg ttcgtgacac aaagaaactt ctacgaaccc cagattatca
     5101 ccaccgacaa caccttcgtg agtggcaatt gtgacgtggt catcggaatc gtgaacaaca
     5161 cagtgtacga ccctctgcaa cctgagctgg actcttttaa ggaagagctg gacaagtact
     5221 ttaaaaacca caccagcccc gatgtggacc tgggcgacat cagtggcatt aacgccagcg
     5281 tggtgaacat ccaaaaggaa atcgacagac tgaacgaggt ggccaagaac ctgaacgagt
     5341 ccctgatcga cctgcaggag ctcggcaaat acgagcaggg atccggatac atccccgagg
     5401 cccccagaga tggccaggcc tacgtgcgga aggacggcga gtgggtactg ctgagcacat
     5461 tcctgggcag atccctggag gtgctgttcc agggcccagg ccatcaccac catcaccacc
     5521 atcatagcgc ctggtcccac ccccagttcg agaagggcgg cggtagtgga gggggcggat
     5581 ctggcggctc agcttggagc cacccccagt tcgaaaagtg ataatgactc gagcgataat
     5641 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     5701 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     5761 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     5821 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     5881 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     5941 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     6001 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     6061 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     6121 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     6181 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6241 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6301 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6361 tgccagcgga tcgatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6421 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7381 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7501 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7681 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7741 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7801 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7861 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7921 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7981 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8041 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8101 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8161 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8221 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8281 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8341 acatttcccc gaaaagtgcc acctgggtc
//
