LOCUS       sequence-363822-        8539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses codon-optimized full length SARS-CoV-2 BA.2.12.1 Spike.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8539)
  AUTHORS   Gong SY, Ding S, Benlarbi M, Chen Y, Vezina D, Marchitto L, 
            Beaudoin-Bussieres G, Goyette G, Bourassa C, Bo Y, Medjahed H, 
            Levade I, Pazgier M, Cote M, Richard J, Prevost J, Finzi A
  TITLE     Temperature Influences the Interaction between SARS-CoV-2 Spike from
            Omicron Subvariants and Human ACE2.
  JOURNAL   Viruses. 2022 Sep 30;14(10):2178. doi: 10.3390/v14102178.
  PUBMED    36298733
REFERENCE   2  (bases 1 to 8539)
  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: "Viruses."; 
            date: "2022-09-30"; volume: "14(10):2178. doi"; pages: " 
            10.3390/v14102178"
COMMENT     Sequence Label: pCAGGS SARS-CoV-2 BA.2.12.1 Spike
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(5560..5579)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5625..5680
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5679..5698)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6041..6057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6041..6057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6054..6076)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6065..6081
                     /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(6089..6095,6096..6113,6114..6119))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6134..6155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6214..6409
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6260..6395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6415..6549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6452..6471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6506..6525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6617..6634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6788..7376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6868..6887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7547..8338,8339..8407))
                     /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     8170..8189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8408..8512)
                     /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 ggggggcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggsggggsgr ggsggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgccaccatg
     1741 ttcgtcttcc tcgtcctcct ccccctcgtc tcctcccagt gcgtcaacct catcacccgc
     1801 acccagagct acaccaactc cttcacccgc ggcgtctact accccgacaa ggtcttccgc
     1861 tcctccgtcc tccactccac ccaggacctc ttcctcccct tcttctccaa cgtcacctgg
     1921 ttccacgcca tccacgtctc cggcaccaac ggcaccaagc gcttcgacaa ccccgtcctc
     1981 cccttcaacg acggcgtcta cttcgcctcc accgagaagt ccaacatcat ccgcggctgg
     2041 atcttcggca ccaccctcga ctccaagacc cagtccctcc tcatcgtcaa caacgccacc
     2101 aacgtcgtca tcaaggtctg cgagttccag ttctgcaacg accccttcct cgacgtctac
     2161 taccacaaga acaacaagtc ctggatggag tccgagttcc gcgtctactc ctccgccaac
     2221 aactgcacct tcgagtacgt ctcccagccc ttcctcatgg acctcgaggg caagcagggc
     2281 aacttcaaga acctccgcga gttcgtcttc aagaacatcg acggctactt caagatctac
     2341 tccaagcaca cccccatcaa cctcggccgc gacctccccc agggcttctc cgccctcgag
     2401 cccctcgtcg acctccccat cggcatcaac atcacccgct tccagaccct cctcgccctc
     2461 caccgctcct acctcacccc cggcgactcc tcctccggct ggaccgccgg cgccgccgcc
     2521 tactacgtcg gctacctcca gccccgcacc ttcctcctca agtacaacga gaacggcacc
     2581 atcaccgacg ccgtcgactg cgccctcgac cccctctccg agaccaagtg caccctcaag
     2641 tccttcaccg tcgagaaggg catctaccag acctccaact tccgcgtcca gcccaccgag
     2701 tccatcgtcc gcttccccaa catcaccaac ctctgcccct tcgacgaggt cttcaacgcc
     2761 acccgcttcg cctccgtcta cgcctggaac cgcaagcgca tctccaactg cgtcgccgac
     2821 tactccgtcc tctacaactt cgcccctttc ttcgccttca agtgctacgg cgtctccccc
     2881 accaagctca acgacctctg cttcaccaac gtctacgccg actccttcgt catccgcggc
     2941 aacgaggtca gccagatcgc ccccggccag accggcaaca tcgccgacta caactacaag
     3001 ctccccgacg acttcaccgg ctgcgtcatc gcctggaact ccaacaagct cgactccaag
     3061 gtcggcggca actacaacta ccagtaccgc ctcttccgca agtccaacct caagcccttc
     3121 gagcgcgaca tctccaccga gatctaccag gccggcaaca aaccctgcaa cggcgtcgcc
     3181 ggcttcaact gctacttccc cctcaggtcc tacggcttca ggcccaccta cggcgtcggc
     3241 caccagccct accgcgtcgt cgtcctctcc ttcgagctcc tccacgcccc cgccaccgtc
     3301 tgcggcccca agaagtccac caacctcgtc aagaacaagt gcgtcaactt caacttcaac
     3361 ggcctcaccg gcaccggcgt cctcaccgag tccaacaaga agttcctccc cttccagcag
     3421 ttcggccgcg acatcgccga caccaccgac gccgtccgcg acccccagac cctcgagatc
     3481 ctcgacatca ccccctgctc cttcggcggc gtctccgtca tcacccccgg caccaacacc
     3541 tccaaccagg tcgccgtcct ctaccagggt gtcaactgca ccgaggtccc cgtcgccatc
     3601 cacgccgacc agctcacccc cacctggcgc gtctactcca ccggctccaa cgtcttccag
     3661 acccgcgccg gctgcctcat cggcgccgag tacgtcaaca actcctacga gtgcgacatc
     3721 cccatcggcg ccggcatctg cgcctcctac cagacccaga ccaagtccca ccgccgcgcc
     3781 cgctccgtcg cctcccagtc catcatcgcc tacaccatgt ccctcggcgc cgagaacctg
     3841 gtcgcctact ccaacaactc catcgccatc cccaccaact tcaccatctc cgtcaccacc
     3901 gagatcctcc ccgtctccat gaccaagacc tccgtcgact gcaccatgta catctgcggc
     3961 gactccaccg agtgctccaa cctcctcctc cagtacggct ccttctgcac ccagctcaag
     4021 cgcgccctca ccggcatcgc cgtcgagcag gacaagaaca cccaggaggt cttcgcccag
     4081 gtcaagcaga tctacaagac cccccccatc aagtacttcg gcggcttcaa cttctcccag
     4141 atcctccccg acccctccaa gccctccaag cgctccttca tcgaggacct cctcttcaac
     4201 aaggtcaccc tcgccgacgc cggcttcatc aagcagtacg gcgactgcct cggcgacatc
     4261 gccgcccgcg acctcatctg cgcccagaag ttcaacggcc tcaccgtcct cccccccctc
     4321 ctcaccgacg agatgatcgc ccagtacacc tccgccctcc tcgccggcac catcacctcc
     4381 ggctggacct tcggcgccgg cgccgccctc cagatcccct tcgccatgca gatggcctac
     4441 cgcttcaacg gcatcggcgt cacccagaac gtcctctacg agaaccagaa gctcatcgcc
     4501 aaccagttca actccgccat cggcaagatc caggactccc tctcctccac cgcctccgcc
     4561 ctcggcaagc tccaggacgt cgtcaaccac aacgcccagg ccctcaacac cctcgtcaag
     4621 cagctctcct ccaagttcgg cgccatctcc tccgtcctca acgacatcct gtcccgcctc
     4681 gacaaggtcg aggccgaggt ccagatcgac cgcctcatca ccggccgcct ccagtccctc
     4741 cagacctacg tcacccagca gctcatccgc gccgccgaga tccgcgcctc cgccaacctc
     4801 gccgccacca agatgtccga gtgcgtcctc ggccagtcca agcgcgtcga cttctgcggc
     4861 aagggctacc acctcatgtc cttcccccag tccgcccccc acggcgtcgt cttcctccac
     4921 gtcacctacg tccccgccca ggagaagaac ttcaccaccg cccccgccat ctgccacgac
     4981 ggcaaggccc acttcccccg cgagggcgtc ttcgtctcca acggcaccca ctggttcgtc
     5041 acccagcgca acttctacga gccccagatc atcaccaccg acaacacctt cgtctccggc
     5101 aactgcgacg tcgtcatcgg catcgtcaac aacaccgtct acgaccccct ccagcccgag
     5161 ctcgactcct tcaaggagga gctcgacaag tacttcaaga accacacctc ccccgacgtc
     5221 gacctcggcg acatctccgg catcaacgcc tccgtcgtca acatccagaa ggagatcgac
     5281 cgcctcaacg aggtcgccaa gaacctcaac gagtccctca tcgacctcca ggagctcggc
     5341 aagtacgagc agtacatcaa gtggccctgg tacatctggc tcggcttcat cgccggcctc
     5401 atcgccatcg tcatggtcac catcatgctc tgctgcatga cctcctgctg ctcctgcctc
     5461 aagggctgct gctcctgcgg ctcctgctgc aagttcgacg aggacgactc cgagcccgtc
     5521 ctcaagggcg tcaagctcca ctacacctaa gctagcagat ctttttccct ctgccaaaaa
     5581 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     5641 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     5701 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     5761 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     5821 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     5881 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     5941 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     6001 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6061 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6121 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6181 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     6241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     6421 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6481 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6541 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6601 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8521 cccgaaaagt gccacctgg
//
