LOCUS       sequence-363833-        5498 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian cell expression of RBD of SARS-CoV-2 spike protein of 
            variant OMICRON.BA.1 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 5498)
  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 5498)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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.1-RBD
FEATURES             Location/Qualifiers
     source          1..5498
                     /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"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2445..2462
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2520..2539)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2585..2640
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2639..2658)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3001..3017)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3001..3017)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3014..3036)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3025..3041
                     /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(3049..3055,3056..3073,3074..3079))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3094..3115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3174..3369
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3220..3355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3375..3509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3412..3431)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3466..3485
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3577..3594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3748..4336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3828..3847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4507..5298,5299..5367))
                     /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     5130..5149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5368..5472)
                     /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 rggcggrgag 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 ggrggggcgc 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 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaaggcc accatgttcg
     1741 tgtttctggt gctgctgcct ctggtgtcca gccagcgggt gcagcctaca gagagcatcg
     1801 tgcgatttcc aaacattacc aacctctgcc ccttcgacga ggtgtttaac gccacaagat
     1861 ttgcctccgt ttacgcctgg aatagaaaga gaatcagcaa ttgtgtggcc gactactccg
     1921 tgctgtataa cctcgcccct ttcttcacct tcaagtgcta cggcgtttcc ccaacaaagc
     1981 tgaatgacct gtgcttcacc aacgtgtacg ccgactcctt cgtaattaga ggcgatgagg
     2041 tgcggcagat cgcaccaggc cagaccggta acatcgctga ctacaactat aagctgcctg
     2101 atgattttac aggctgcgtg atcgcctgga actctaacaa gctggatagc aaggtgagcg
     2161 gcaactacaa ctacctgtac cggctgtttc gcaagtctaa cctgaaacct ttcgagagag
     2221 acatctccac agagatctac caggccggta acaagccttg taacggggtg gccggcttca
     2281 actgttactt ccctctgaga agctacagct tccggcctac ctatggagtc ggccaccagc
     2341 cataccgggt ggtcgtgctg tccttcgagt tactccacgc ccccgccacc gtctgcggtc
     2401 ctaagaagtc caccaatctg gttaagaaca aatgcgtgaa cttccaccat caccatcacc
     2461 attgataaaa ttcgagctcg cggccgcatc gatcttaagt cgcgactcga gctagcagat
     2521 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     2581 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     2641 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     2701 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     2761 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     2821 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     2881 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     2941 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     3001 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3061 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3121 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     3181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3361 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3481 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     3541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5461 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
