LOCUS       sequence-319569-        8358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SARS-CoV-2 Spike protein with furin site mutated to HRV3C site 
            (S-HRV3C variant) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8358)
  AUTHORS   Gobeil SM, Janowska K, McDowell S, Mansouri K, Parks R, Manne K, 
            Stalls V, Kopp MF, Henderson R, Edwards RJ, Haynes BF, Acharya P
  TITLE     D614G Mutation Alters SARS-CoV-2 Spike Conformation and Enhances 
            Protease Cleavage at the S1/S2 Junction.
  JOURNAL   Cell Rep. 2021 Jan 12;34(2):108630. doi: 
            10.1016/j.celrep.2020.108630. Epub 2020 Dec 26.
  PUBMED    33417835
REFERENCE   2  (bases 1 to 8358)
  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: "2021-01-12"; volume: "34(2):108630. doi"; pages: " 
            10.1016/j.celrep.2020.108630. Epub 2020 Dec 26"
COMMENT     Sequence Label: p-alpha-H-S-HRV3C
FEATURES             Location/Qualifiers
     source          1..8358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             3805..3828
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5497..5520
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5527..5550
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5590..5613
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5715..5734)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5780..5835
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5834..5853)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6194..6210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6194..6210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6207..6229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6218..6234
                     /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(6242..6248,6249..6266,6267..6272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6287..6308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6437..6454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6608..7196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6688..6707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7367..8158,8159..8227))
                     /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     7990..8009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8228..8332)
                     /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 tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gcgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattgcg gccgtctcag
     1741 gccgagttcg gtaccgccac catgttcgtg ttcctggtgc tcctgcctct ggtgagcagc
     1801 cagtgcgtga acctgaccac ccgaacccag ctcccaccag cctacaccaa cagctttaca
     1861 cggggcgtgt actaccctga caaggtgttc agatctagcg tcctgcacag cactcaggac
     1921 ctcttcctgc cgttcttcag caacgtgaca tggttccacg ccatccacgt gagcggcaca
     1981 aacggaacca agcggtttga taaccccgtc ctgccattca atgatggagt ttacttcgcc
     2041 agtaccgaga agagtaacat catccggggc tggatcttcg gcaccaccct ggatagcaaa
     2101 acacagagcc tcctgatcgt gaacaatgcc acgaacgtcg tgatcaaggt gtgcgagttc
     2161 cagttttgca atgatccttt cctgggtgtg tactaccaca agaacaacaa gagctggatg
     2221 gaaagcgagt tcagagtcta cagcagcgcc aacaactgca cattcgagta cgtctctcag
     2281 ccttttctga tggaccttga ggggaaacaa ggcaacttca agaacctgag agaattcgtg
     2341 ttcaagaaca tcgacggcta cttcaaaatc tactccaagc acacacccat caacctggtc
     2401 cgggacctcc ctcagggctt cagcgccctg gaacccctgg tcgacctgcc cataggcatc
     2461 aacataacgc ggttccaaac cctgctggcc ctgcatagat cctacctgac tcctggcgac
     2521 agcagcagcg gatggaccgc cggagctgca gcctactatg tgggctacct gcaacctaga
     2581 accttcctgc tgaagtacaa cgagaacggc acaatcacag acgccgtcga ctgcgccctg
     2641 gaccctctct ctgagacaaa gtgcaccctg aagtccttca ccgtggaaaa gggcatctac
     2701 cagaccagca acttccgggt gcagcctaca gagagcatcg tgcgatttcc aaacattacc
     2761 aacctctgcc ccttcggcga ggtgtttaac gccacaagat ttgcctccgt ttacgcctgg
     2821 aatagaaaga gaatcagcaa ttgtgtggcc gactactccg tgctgtataa cagcgcctct
     2881 ttcagcacct tcaagtgcta cggcgtttcc ccaacaaagc tgaatgacct gtgcttcacc
     2941 aacgtgtacg ccgactcctt cgtaattaga ggcgatgagg tgcggcagat cgcaccaggc
     3001 cagaccggta agatcgctga ctacaactat aagctgcctg atgattttac aggctgcgtg
     3061 atcgcctgga actctaacaa cctggatagc aaggtgggcg gcaactacaa ctacctgtac
     3121 cggctgtttc gcaagtctaa cctgaaacct ttcgagagag acatctccac agagatctac
     3181 caggccggtt ctacaccttg taacggggtg gaaggcttca actgttactt ccctctgcaa
     3241 agctacggct tccagcctac caatggagtc ggctaccagc cataccgggt ggtcgtgctg
     3301 tccttcgagt tactccacgc ccccgccacc gtctgcggtc ctaagaagtc caccaatctg
     3361 gttaagaaca aatgcgtgaa cttcaacttc aacggcctga ccgggaccgg cgtgctgacc
     3421 gaaagcaaca aaaagttcct ccccttccag cagttcggcc gtgatatcgc tgacaccaca
     3481 gatgccgtca gagatccaca gaccctggaa atcctggata ttacaccctg ctccttcgga
     3541 ggagtttctg tgatcacccc cgggaccaat accagcaacc aggtggctgt gctgtaccaa
     3601 gatgttaact gcaccgaggt tcctgtggcc atccacgccg atcagctgac acctacttgg
     3661 agagtgtact ccactggctc caatgtgttc cagaccaggg ccggatgtct gatcggcgcc
     3721 gagcacgtga ataacagtta cgagtgcgac atccctatcg gcgccggcat ctgtgccagc
     3781 taccagaccc agacaaacag ccctctggag gtgctgttcc agggcccatc tgtagctagc
     3841 cagagcatca tcgcctacac catgagcctg ggcgcagaga acagcgtggc ctattccaac
     3901 aactctatcg ccattcccac caactttaca attagcgtca caacagagat cctgcccgtg
     3961 agcatgacca agaccagcgt ggactgtaca atgtacatct gtggcgacag cactgaatgc
     4021 agcaacctgc tgctgcaata cggctccttt tgcacccaac tgaaccgggc gctgaccgga
     4081 atcgccgtgg aacaggacaa aaatacccag gaggtgttcg cccaagtgaa gcagatctac
     4141 aagaccccac ctatcaagga cttcggcggc tttaacttta gccagattct ccctgatcct
     4201 tctaagccta gcaagcggag ctttatcgag gatctgctgt tcaacaaggt caccctggcc
     4261 gatgccggct ttatcaaaca gtatggcgat tgcctgggcg acatagccgc cagagatctg
     4321 atctgcgccc agaaattcaa cggcctgaca gttctcccac ctctgctgac cgacgagatg
     4381 atcgctcagt acacctctgc cctgctggct ggcaccatca catctgggtg gacatttggc
     4441 gccggcgccg ccctgcagat cccctttgcc atgcagatgg cctatagatt caacggaatc
     4501 ggcgtgaccc agaacgtgct gtatgaaaac cagaagctga tcgctaacca gttcaattct
     4561 gccatcggca agatccagga ctccctctcc tctaccgcca gcgccctggg caaactgcag
     4621 gacgtggtga atcagaacgc ccaagccctg aacaccctgg tgaagcagct cagcagcaat
     4681 tttggcgcca tcagctctgt gctgaacgat atcctgtcta gactggacaa ggtggaagcc
     4741 gaagtccaga tcgatagact gatcacaggc agactgcagt ccctgcaaac ctacgtgacc
     4801 caacagctga tcagggccgc tgaaataaga gccagcgcca atctcgccgc taccaagatg
     4861 tccgagtgtg tgctgggaca gtctaaacgc gttgacttct gcggcaaagg ctatcacctg
     4921 atgagcttcc cccagagcgc gccgcacggc gtggtgttcc tgcatgtgac atacgtgcct
     4981 gcccaagaga agaatttcac aaccgcccct gccatctgcc acgacggcaa ggcccacttc
     5041 ccaagagagg gcgttttcgt ttccaatggc acacactggt tcgtgacaca aagaaacttc
     5101 tacgaacccc agattatcac caccgacaac accttcgtga gtggcaattg tgacgtggtc
     5161 atcggaatcg tgaacaacac agtgtacgac cctctgcaac ctgagctgga ctcttttaag
     5221 gaagagctgg acaagtactt taaaaaccac accagccccg atgtggacct gggcgacatc
     5281 agtggcatta acgccagcgt ggtgaacatc caaaaggaaa tcgacagact gaacgaggtg
     5341 gccaagaacc tgaacgagtc cctgatcgac ctgcaggagc tcggcaaata cgagcaggga
     5401 tccggataca tccccgaggc ccccagagat ggccaggcct acgtgcggaa ggacggcgag
     5461 tgggtactgc tgagcacatt cctgggcaga tccggccatc accaccatca ccaccatcat
     5521 agcgcctggt cccaccccca gttcgagaag ggcggcggta gtggaggggg cggatctggc
     5581 ggctcagctt ggagccaccc ccagttcgaa aagtgataat gactcgagcg ataattcact
     5641 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     5701 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     5761 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     5821 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     5881 gagtatttgg tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg
     5941 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     6001 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     6061 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     6121 actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctgc agcccaagct
     6181 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6241 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6301 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6361 ggatcgatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6421 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6781 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     6841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7021 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7321 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7381 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7441 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7501 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7561 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7621 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7681 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7741 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7801 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7861 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7921 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7981 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8041 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8101 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8161 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8221 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8341 cccgaaaagt gccacctg
//
