LOCUS       sequence-324833-        8363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SARS-CoV-2 Spike protein with furin site mutated to HRV3C site and 
            D614G mutation (S-HRV3C-D614G variant) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8363)
  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 8363)
  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/D614G
FEATURES             Location/Qualifiers
     source          1..8363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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"
     CDS             3808..3831
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5500..5523
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5530..5553
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5593..5616
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5718..5737)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5783..5838
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5837..5856)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6199..6215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6199..6215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6212..6234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6223..6239
                     /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(6247..6253,6254..6271,6272..6277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6292..6313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6442..6459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6613..7201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6693..6712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7372..8163,8164..8232))
                     /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     7995..8014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8233..8337)
                     /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 ggggggsgsg ccvggsgggg sggggsgggg sgrggggsgg
      541 ggsggggsgr ggsggrgrgg kgsggsggcr 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 kggcggcrgg kgggggkgcc gggcggggcg
     1261 gggccgcckc gggcsggggr gggskcgggg grggggcgcg gcggcccccg grgcgccggc
     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 ctagcgcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt gcggccgtct
     1741 caggccgagt tcggtaccgc caccatgttc gtgttcctgg tgctcctgcc tctggtgagc
     1801 agccagtgcg tgaacctgac cacccgaacc cagctcccac cagcctacac caacagcttt
     1861 acacggggcg tgtactaccc tgacaaggtg ttcagatcta gcgtcctgca cagcactcag
     1921 gacctcttcc tgccgttctt cagcaacgtg acatggttcc acgccatcca cgtgagcggc
     1981 acaaacggaa ccaagcggtt tgataacccc gtcctgccat tcaatgatgg agtttacttc
     2041 gccagtaccg agaagagtaa catcatccgg ggctggatct tcggcaccac cctggatagc
     2101 aaaacacaga gcctcctgat cgtgaacaat gccacgaacg tcgtgatcaa ggtgtgcgag
     2161 ttccagtttt gcaatgatcc tttcctgggt gtgtactacc acaagaacaa caagagctgg
     2221 atggaaagcg agttcagagt ctacagcagc gccaacaact gcacattcga gtacgtctct
     2281 cagccttttc tgatggacct tgaggggaaa caaggcaact tcaagaacct gagagaattc
     2341 gtgttcaaga acatcgacgg ctacttcaaa atctactcca agcacacacc catcaacctg
     2401 gtccgggacc tccctcaggg cttcagcgcc ctggaacccc tggtcgacct gcccataggc
     2461 atcaacataa cgcggttcca aaccctgctg gccctgcata gatcctacct gactcctggc
     2521 gacagcagca gcggatggac cgccggagct gcagcctact atgtgggcta cctgcaacct
     2581 agaaccttcc tgctgaagta caacgagaac ggcacaatca cagacgccgt cgactgcgcc
     2641 ctggaccctc tctctgagac aaagtgcacc ctgaagtcct tcaccgtgga aaagggcatc
     2701 taccagacca gcaacttccg ggtgcagcct acagagagca tcgtgcgatt tccaaacatt
     2761 accaacctct gccccttcgg cgaggtgttt aacgccacaa gatttgcctc cgtttacgcc
     2821 tggaatagaa agagaatcag caattgtgtg gccgactact ccgtgctgta taacagcgcc
     2881 tctttcagca ccttcaagtg ctacggcgtt tccccaacaa agctgaatga cctgtgcttc
     2941 accaacgtgt acgccgactc cttcgtaatt agaggcgatg aggtgcggca gatcgcacca
     3001 ggccagaccg gtaagatcgc tgactacaac tataagctgc ctgatgattt tacaggctgc
     3061 gtgatcgcct ggaactctaa caacctggat agcaaggtgg gcggcaacta caactacctg
     3121 taccggctgt ttcgcaagtc taacctgaaa cctttcgaga gagacatctc cacagagatc
     3181 taccaggccg gttctacacc ttgtaacggg gtggaaggct tcaactgtta cttccctctg
     3241 caaagctacg gcttccagcc taccaatgga gtcggctacc agccataccg ggtggtcgtg
     3301 ctgtccttcg agttactcca cgcccccgcc accgtctgcg gtcctaagaa gtccaccaat
     3361 ctggttaaga acaaatgcgt gaacttcaac ttcaacggcc tgaccgggac cggcgtgctg
     3421 accgaaagca acaaaaagtt cctccccttc cagcagttcg gccgtgatat cgctgacacc
     3481 acagatgccg tcagagatcc acagaccctg gaaatcctgg atattacacc ctgctccttc
     3541 ggaggagttt ctgtgatcac ccccgggacc aataccagca accaggtggc tgtgctgtac
     3601 caaggtgtta actgcaccga ggttcctgtg gccatccacg ccgatcagct gacacctact
     3661 tggagagtgt actccactgg ctccaatgtg ttccagacca gggccggatg tctgatcggc
     3721 gccgagcacg tgaataacag ttacgagtgc gacatcccta tcggcgccgg catctgtgcc
     3781 agctaccaga cccagacaaa cagccctctg gaggtgctgt tccagggccc atctgtagct
     3841 agccagagca tcatcgccta caccatgagc ctgggcgcag agaacagcgt ggcctattcc
     3901 aacaactcta tcgccattcc caccaacttt acaattagcg tcacaacaga gatcctgccc
     3961 gtgagcatga ccaagaccag cgtggactgt acaatgtaca tctgtggcga cagcactgaa
     4021 tgcagcaacc tgctgctgca atacggctcc ttttgcaccc aactgaaccg ggcgctgacc
     4081 ggaatcgccg tggaacagga caaaaatacc caggaggtgt tcgcccaagt gaagcagatc
     4141 tacaagaccc cacctatcaa ggacttcggc ggctttaact ttagccagat tctccctgat
     4201 ccttctaagc ctagcaagcg gagctttatc gaggatctgc tgttcaacaa ggtcaccctg
     4261 gccgatgccg gctttatcaa acagtatggc gattgcctgg gcgacatagc cgccagagat
     4321 ctgatctgcg cccagaaatt caacggcctg acagttctcc cacctctgct gaccgacgag
     4381 atgatcgctc agtacacctc tgccctgctg gctggcacca tcacatctgg gtggacattt
     4441 ggcgccggcg ccgccctgca gatccccttt gccatgcaga tggcctatag attcaacgga
     4501 atcggcgtga cccagaacgt gctgtatgaa aaccagaagc tgatcgctaa ccagttcaat
     4561 tctgccatcg gcaagatcca ggactccctc tcctctaccg ccagcgccct gggcaaactg
     4621 caggacgtgg tgaatcagaa cgcccaagcc ctgaacaccc tggtgaagca gctcagcagc
     4681 aattttggcg ccatcagctc tgtgctgaac gatatcctgt ctagactgga caaggtggaa
     4741 gccgaagtcc agatcgatag actgatcaca ggcagactgc agtccctgca aacctacgtg
     4801 acccaacagc tgatcagggc cgctgaaata agagccagcg ccaatctcgc cgctaccaag
     4861 atgtccgagt gtgtgctggg acagtctaaa cgcgttgact tctgcggcaa aggctatcac
     4921 ctgatgagct tcccccagag cgcgccgcac ggcgtggtgt tcctgcatgt gacatacgtg
     4981 cctgcccaag agaagaattt cacaaccgcc cctgccatct gccacgacgg caaggcccac
     5041 ttcccaagag agggcgtttt cgtttccaat ggcacacact ggttcgtgac acaaagaaac
     5101 ttctacgaac cccagattat caccaccgac aacaccttcg tgagtggcaa ttgtgacgtg
     5161 gtcatcggaa tcgtgaacaa cacagtgtac gaccctctgc aacctgagct ggactctttt
     5221 aaggaagagc tggacaagta ctttaaaaac cacaccagcc ccgatgtgga cctgggcgac
     5281 atcagtggca ttaacgccag cgtggtgaac atccaaaagg aaatcgacag actgaacgag
     5341 gtggccaaga acctgaacga gtccctgatc gacctgcagg agctcggcaa atacgagcag
     5401 ggatccggat acatccccga ggcccccaga gatggccagg cctacgtgcg gaaggacggc
     5461 gagtgggtac tgctgagcac attcctgggc agatccggcc atcaccacca tcaccaccat
     5521 catagcgcct ggtcccaccc ccagttcgag aagggcggcg gtagtggagg gggcggatct
     5581 ggcggctcag cttggagcca cccccagttc gaaaagtgat aatgactcga gcgataattc
     5641 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     5701 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     5761 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     5821 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     5881 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
     5941 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     6001 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
     6061 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     6121 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc
     6181 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6241 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6301 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6361 ccagcggatc gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7021 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8341 atttccccga aaagtgccac ctg
//
