LOCUS       sequence-324845-        8367 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 soluble Spike protein with a D614G mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8367)
  AUTHORS   Yurkovetskiy L, Wang X, Pascal KE, Tomkins-Tinch C, Nyalile TP, Wang
            Y, Baum A, Diehl WE, Dauphin A, Carbone C, Veinotte K, Egri SB, 
            Schaffner SF, Lemieux JE, Munro JB, Rafique A, Barve A, Sabeti PC, 
            Kyratsous CA, Dudkina NV, Shen K, Luban J
  TITLE     Structural and Functional Analysis of the D614G SARS-CoV-2 Spike 
            Protein Variant.
  JOURNAL   Cell. 2020 Sep 15. pii: S0092-8674(20)31229-0. doi: 
            10.1016/j.cell.2020.09.032.
  PUBMED    32991842
REFERENCE   2  (bases 1 to 8367)
  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. 2020 
            Sep 15. pii: S0092-8674(20)31229-0. doi: 
            10.1016/j.cell.2020.09.032."
COMMENT     Sequence Label: pCAGGS D614G soluble Spike
FEATURES             Location/Qualifiers
     source          1..8367
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          488..1504
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1428..1450
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1512..1531
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5199..5216
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5310..5327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5385..5404)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5450..5505
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5504..5523)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5866..5882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5866..5882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5879..5901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5890..5906
                     /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(5914..5920,5921..5938,5939..5944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5959..5980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6039..6234
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6085..6220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6240..6374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6277..6296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6331..6350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     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 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccacggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      481 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      541 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      601 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      661 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
      721 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
      781 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
      841 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
      901 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
      961 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
     1021 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
     1081 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
     1141 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     1201 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
     1261 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     1321 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1381 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1441 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1501 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aaggccacca
     1561 tgttcgtgtt tctggtgctg ctgcctctgg tgtccagcca gtgtgtgaac ctgaccacaa
     1621 gaacccagct gcctccagcc tacaccaaca gctttaccag aggcgtgtac taccccgaca
     1681 aggtgttcag atccagcgtg ctgcactcta cccaggacct gttcctgcct ttcttcagca
     1741 acgtgacctg gttccacgcc atccacgtgt ccggcaccaa tggcaccaag agattcgaca
     1801 accccgtgct gcccttcaac gacggggtgt actttgccag caccgagaag tccaacatca
     1861 tcagaggctg gatcttcggc accacactgg acagcaagac ccagagcctg ctgatcgtga
     1921 acaacgccac caacgtggtc atcaaagtgt gcgagttcca gttctgcaac gaccccttcc
     1981 tgggcgtcta ctatcacaag aacaacaaga gctggatgga aagcgagttc cgggtgtaca
     2041 gcagcgccaa caactgcacc ttcgagtacg tgtcccagcc tttcctgatg gacctggaag
     2101 gcaagcaggg caacttcaag aacctgcgcg agttcgtgtt caagaacatc gacggctact
     2161 tcaagatcta cagcaagcac acccctatca acctcgtgcg ggatctgcct cagggcttct
     2221 ctgctctgga acccctggtg gatctgccca tcggcatcaa catcacccgg tttcagacac
     2281 tgctggccct gcacagaagc tacctgacac ctggcgatag cagcagcgga tggacagctg
     2341 gtgccgccgc ttactatgtg ggctacctgc agcctagaac ctttctgctg aagtacaacg
     2401 agaacggcac catcaccgac gccgtggatt gtgctctgga tcctctgagc gagacaaagt
     2461 gcaccctgaa gtccttcacc gtggaaaagg gcatctacca gaccagcaac ttccgggtgc
     2521 agcccaccga atccatcgtg cggttcccca atatcaccaa tctgtgcccc ttcggcgagg
     2581 tgttcaatgc caccagattc gcctctgtgt acgcctggaa ccggaagcgg atcagcaatt
     2641 gcgtggccga ctactccgtg ctgtacaact ccgccagctt cagcaccttc aagtgctacg
     2701 gcgtgtcccc taccaagctg aacgacctgt gcttcacaaa cgtgtacgcc gacagcttcg
     2761 tgatccgggg agatgaagtg cggcagattg cccctggaca gacaggcaag atcgccgact
     2821 acaactacaa gctgcccgac gacttcaccg gctgtgtgat tgcctggaac agcaacaacc
     2881 tggactccaa agtcggcggc aactacaatt acctgtaccg gctgttccgg aagtccaatc
     2941 tgaagccctt cgagcgggac atctccaccg agatctatca ggccggcagc accccttgta
     3001 acggcgtgga aggcttcaac tgctacttcc cactgcagtc ctacggcttt cagcccacaa
     3061 atggcgtggg ctatcagccc tacagagtgg tggtgctgag cttcgaactg ctgcatgccc
     3121 ctgccacagt gtgcggccct aagaaaagca ccaatctcgt gaagaacaaa tgcgtgaact
     3181 tcaacttcaa cggcctgacc ggcaccggcg tgctgacaga gagcaacaag aagttcctgc
     3241 cattccagca gtttggccgg gatatcgccg ataccacaga cgccgttaga gatccccaga
     3301 cactggaaat cctggacatc accccttgca gcttcggcgg agtgtctgtg atcacccctg
     3361 gcaccaacac cagcaatcag gtggcagtgc tgtaccaggg cgtgaactgt accgaagtgc
     3421 ccgtggccat tcacgccgat cagctgacac ctacatggcg ggtgtactcc accggcagca
     3481 atgtgtttca gaccagagcc ggctgtctga tcggagccga gcacgtgaac aatagctacg
     3541 agtgcgacat ccccatcggc gctggcatct gtgccagcta ccagacacag acaaacagcc
     3601 ccgcctctgt ggccagccag agcatcattg cctacacaat gtctctgggc gccgagaaca
     3661 gcgtggccta ctccaacaac tctatcgcta tccccaccaa cttcaccatc agcgtgacca
     3721 cagagatcct gcctgtgtcc atgaccaaga ccagcgtgga ctgcaccatg tacatctgcg
     3781 gcgattccac cgagtgctcc aacctgctgc tgcagtacgg cagcttctgc acccagctga
     3841 atagagccct gacagggatc gccgtggaac aggacaagaa cacccaagag gtgttcgccc
     3901 aagtgaagca gatctacaag acccctccta tcaaggactt cggcggcttc aatttcagcc
     3961 agattctgcc cgatcctagc aagcccagca agcggagctt catcgaggac ctgctgttca
     4021 acaaagtgac actggccgac gccggcttca tcaagcagta tggcgattgt ctgggcgaca
     4081 ttgccgccag ggatctgatt tgcgcccaga agtttaacgg actgacagtg ctgcctcctc
     4141 tgctgaccga tgagatgatc gcccagtaca catctgccct gctggccggc acaatcacaa
     4201 gcggctggac atttggagct ggcgccgctc tgcagatccc ctttgctatg cagatggcct
     4261 accggttcaa cggcatcgga gtgacccaga atgtgctgta cgagaaccag aagctgatcg
     4321 ccaaccagtt caacagcgcc atcggcaaga tccaggacag cctgagcagc acagcaagcg
     4381 ccctgggaaa gctgcaggac gtggtcaacc agaatgccca ggcactgaac accctggtca
     4441 agcagctgtc ctccaacttc ggcgccatca gctctgtgct gaacgatatc ctgagcagac
     4501 tggaccctcc tgaagccgag gtgcagatcg acagactgat caccggaagg ctgcagtccc
     4561 tgcagaccta cgttacccag cagctgatca gagccgccga gattagagcc tctgccaatc
     4621 tggccgccac caagatgtct gagtgtgtgc tgggccagag caagagagtg gacttttgcg
     4681 gcaagggcta ccacctgatg agcttccctc agtctgcccc tcacggcgtg gtgtttctgc
     4741 acgtgacata cgtgcccgct caagagaaga atttcaccac cgctccagcc atctgccacg
     4801 acggcaaagc ccactttcct agagaaggcg tgttcgtgtc caacggcacc cattggttcg
     4861 tgacccagcg gaacttctac gagccccaga tcatcaccac cgacaacacc ttcgtgtctg
     4921 gcaactgcga cgtcgtgatc ggcattgtga acaataccgt gtacgaccct ctgcagcccg
     4981 agctggacag cttcaaagag gaactggata agtactttaa gaaccacaca agccccgacg
     5041 tggacctggg cgatatcagc ggaatcaatg ccagcgtcgt gaacatccag aaagagatcg
     5101 accggctgaa cgaggtggcc aagaatctga acgagagcct gatcgacctg caagaactgg
     5161 ggaagtacga gcagtacatc aagtggccca gcggccgctt ggtcccacgt ggctcacccg
     5221 gatctggata catcccggag gcccctaggg acggtcaagc ttacgtgaga aaggacggcg
     5281 aatgggttct gctgtcgacc ttcttgggac atcatcatca tcatcactaa tgaaattcga
     5341 gctcgcggcc gcatcgatct taagtcgcga ctcgagctag cagatctttt tccctctgcc
     5401 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     5461 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     5521 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     5581 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     5641 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     5701 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     5761 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     5821 ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     5881 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5941 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6001 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     6061 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     6241 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6301 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6361 atcatgtctg 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 ctgggtc
//
