LOCUS       sequence-354264-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of SARS-CoV-2 Spike protein variant 
            OMICRON with furin side replaced by GSAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  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 8349)
  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-GSAS-OMICRON
FEATURES             Location/Qualifiers
     source          1..8349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..642
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          643..1659
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1583..1605
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5458..5481
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5485..5508
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5515..5538
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5578..5601
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5703..5722)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5768..5823
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5822..5841)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6184..6200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6184..6200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6197..6219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6208..6224
                     /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(6232..6238,6239..6256,6257..6262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6277..6298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6427..6444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6598..7186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6678..6697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7357..8148,8149..8217))
                     /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     7980..7999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8218..8322)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg gggggggcgg ggcgaggggc ggggcggggc gaggcggaga
      541 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      601 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      661 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      721 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      781 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      841 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
      901 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
      961 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     1021 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     1081 gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct
     1141 gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg
     1201 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
     1261 gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
     1321 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
     1381 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     1441 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
     1501 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     1561 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagcgcc
     1621 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     1681 gttgtgctgt ctcatcattt tggcaaagaa ttgcggccgt ctcaggccga gttcggtacc
     1741 gccaccatgt tcgtgttcct ggtgctcctg cctctggtga gcagccagtg cgtgaacctg
     1801 accacccgaa cccagctccc accagcctac accaacagct ttacacgggg cgtgtactac
     1861 cctgacaagg tgttcagatc tagcgtcctg cacagcactc aggacctctt cctgccgttc
     1921 ttcagcaacg tgacatggtt ccacgtcatc agcggcacaa acggaaccaa gcggtttgat
     1981 aaccccgtcc tgccattcaa tgatggagtt tacttcgcca gtatcgagaa gagtaacatc
     2041 atccggggct ggatcttcgg caccaccctg gatagcaaaa cacagagcct cctgatcgtg
     2101 aacaatgcca cgaacgtcgt gatcaaggtg tgcgagttcc agttttgcaa tgatcctttc
     2161 ctggaccaca agaacaacaa gagctggatg gaaagcgagt tcagagtcta cagcagcgcc
     2221 aacaactgca cattcgagta cgtctctcag ccttttctga tggaccttga ggggaaacaa
     2281 ggcaacttca agaacctgag agaattcgtg ttcaagaaca tcgacggcta cttcaaaatc
     2341 tactccaagc acacacccat catcgtccgg gagcctgaag acctccctca gggcttcagc
     2401 gccctggaac ccctggtcga cctgcccata ggcatcaaca taacgcggtt ccaaaccctg
     2461 ctggccctgc atagatccta cctgactcct ggcgacagca gcagcggatg gaccgccgga
     2521 gctgcagcct actatgtggg ctacctgcaa cctagaacct tcctgctgaa gtacaacgag
     2581 aacggcacaa tcacagacgc cgtcgactgc gccctggacc ctctctctga gacaaagtgc
     2641 accctgaagt ccttcaccgt ggaaaagggc atctaccaga ccagcaactt ccgggtgcag
     2701 cctacagaga gcatcgtgcg atttccaaac attaccaacc tctgcccctt cgacgaggtg
     2761 tttaacgcca caagatttgc ctccgtttac gcctggaata gaaagagaat cagcaattgt
     2821 gtggccgact actccgtgct gtataacctc gcccctttct tcaccttcaa gtgctacggc
     2881 gtttccccaa caaagctgaa tgacctgtgc ttcaccaacg tgtacgccga ctccttcgta
     2941 attagaggcg atgaggtgcg gcagatcgca ccaggccaga ccggtaacat cgctgactac
     3001 aactataagc tgcctgatga ttttacaggc tgcgtgatcg cctggaactc taacaagctg
     3061 gatagcaagg tgagcggcaa ctacaactac ctgtaccggc tgtttcgcaa gtctaacctg
     3121 aaacctttcg agagagacat ctccacagag atctaccagg ccggtaacaa gccttgtaac
     3181 ggggtggccg gcttcaactg ttacttccct ctgagaagct acagcttccg gcctacctat
     3241 ggagtcggcc accagccata ccgggtggtc gtgctgtcct tcgagttact ccacgccccc
     3301 gccaccgtct gcggtcctaa gaagtccacc aatctggtta agaacaaatg cgtgaacttc
     3361 aacttcaacg gcctgaaggg gaccggcgtg ctgaccgaaa gcaacaaaaa gttcctcccc
     3421 ttccagcagt tcggccgtga tatcgctgac accacagatg ccgtcagaga tccacagacc
     3481 ctggaaatcc tggatattac accctgctcc ttcggaggag tttctgtgat cacccccggg
     3541 accaatacca gcaaccaggt ggctgtgctg taccaaggtg ttaactgcac cgaggttcct
     3601 gtggccatcc acgccgatca gctgacacct acttggagag tgtactccac tggctccaat
     3661 gtgttccaga ccagggccgg atgtctgatc ggcgccgagt acgtgaataa cagttacgag
     3721 tgcgacatcc ctatcggcgc cggcatctgt gccagctacc agacccagac aaagagccat
     3781 gggtctgctt cctctgtagc tagccagagc atcatcgcct acaccatgag cctgggcgca
     3841 gagaacagcg tggcctattc caacaactct atcgccattc ccaccaactt tacaattagc
     3901 gtcacaacag agatcctgcc cgtgagcatg accaagacca gcgtggactg tacaatgtac
     3961 atctgtggcg acagcactga atgcagcaac ctgctgctgc aatacggctc cttttgcacc
     4021 caactgaagc gggcgctgac cggaatcgcc gtggaacagg acaaaaatac ccaggaggtg
     4081 ttcgcccaag tgaagcagat ctacaagacc ccacctatca agtacttcgg cggctttaac
     4141 tttagccaga ttctccctga tccttctaag cctagcaagc ggagctttat cgaggatctg
     4201 ctgttcaaca aggtcaccct ggccgatgcc ggctttatca aacagtatgg cgattgcctg
     4261 ggcgacatag ccgccagaga tctgatctgc gcccagaaat tcaagggcct gacagttctc
     4321 ccacctctgc tgaccgacga gatgatcgct cagtacacct ctgccctgct ggctggcacc
     4381 atcacatctg ggtggacatt tggcgccggc gccgccctgc agatcccctt tgccatgcag
     4441 atggcctata gattcaacgg aatcggcgtg acccagaacg tgctgtatga aaaccagaag
     4501 ctgatcgcta accagttcaa ttctgccatc ggcaagatcc aggactccct ctcctctacc
     4561 gccagcgccc tgggcaaact gcaggacgtg gtgaatcaca acgcccaagc cctgaacacc
     4621 ctggtgaagc agctcagcag caagtttggc gccatcagct ctgtgctgaa cgatatcttc
     4681 tctagactgg acaaggtgga agccgaagtc cagatcgata gactgatcac aggcagactg
     4741 cagtccctgc aaacctacgt gacccaacag ctgatcaggg ccgctgaaat aagagccagc
     4801 gccaatctcg ccgctaccaa gatgtccgag tgtgtgctgg gacagtctaa acgcgttgac
     4861 ttctgcggca aaggctatca cctgatgagc ttcccccaga gcgcgccgca cggcgtggtg
     4921 ttcctgcatg tgacatacgt gcctgcccaa gagaagaatt tcacaaccgc ccctgccatc
     4981 tgccacgacg gcaaggccca cttcccaaga gagggcgttt tcgtttccaa tggcacacac
     5041 tggttcgtga cacaaagaaa cttctacgaa ccccagatta tcaccaccga caacaccttc
     5101 gtgagtggca attgtgacgt ggtcatcgga atcgtgaaca acacagtgta cgaccctctg
     5161 caacctgagc tggactcttt taaggaagag ctggacaagt actttaaaaa ccacaccagc
     5221 cccgatgtgg acctgggcga catcagtggc attaacgcca gcgtggtgaa catccaaaag
     5281 gaaatcgaca gactgaacga ggtggccaag aacctgaacg agtccctgat cgacctgcag
     5341 gagctcggca aatacgagca gggatccgga tacatccccg aggcccccag agatggccag
     5401 gcctacgtgc ggaaggacgg cgagtgggta ctgctgagca cattcctggg cagatccctg
     5461 gaggtgctgt tccagggccc aggccatcac caccatcacc accatcatag cgcctggtcc
     5521 cacccccagt tcgagaaggg cggcggtagt ggagggggcg gatctggcgg ctcagcttgg
     5581 agccaccccc agttcgaaaa gtgataatga ctcgagcgat aattcactcc tcaggtgcag
     5641 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     5701 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     5761 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     5821 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     5881 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     5941 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     6001 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     6061 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     6121 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
     6181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatcgatcc
     6361 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6421 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6481 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6541 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6601 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6661 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6721 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6781 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6841 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6901 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6961 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7021 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7081 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7141 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7201 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7261 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7321 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7381 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7441 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7501 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7561 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7621 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7681 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7741 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7801 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7861 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7921 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7981 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8041 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8101 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8161 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8221 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8281 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8341 gccacctgg
//
