LOCUS       sequence-324846-        8810 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 soluble Spike protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8810)
  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 8810)
  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 D614 soluble Spike
FEATURES             Location/Qualifiers
     source          1..8810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          278..1294
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1218..1240
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1302..1321
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4989..5006
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5100..5117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5175..5194)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5240..5295
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5294..5313)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5656..5672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5656..5672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5669..5691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5680..5696
                     /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(5704..5710,5711..5728,5729..5734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5749..5770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5829..6024
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5875..6010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6030..6164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6067..6086)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6121..6140
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6232..6249)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6403..6991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6483..6502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7162..7953,7954..8022))
                     /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     7785..7804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8023..8127)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8158..8537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggggggg gggggggggs sssssscssc ssrccsccsc cccccccccc cccccanrtt
       61 ttgktttttn tttktttttt aatkttttkg gggggggggg gggggggggg gggggggggg
      121 gcgcgcgccr ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
      181 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      241 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
      301 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      361 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
      421 ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg
      481 ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      541 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
      601 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
      661 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
      721 tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca
      781 cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg
      841 cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg
      901 ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg
      961 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     1021 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
     1081 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     1141 ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg
     1201 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
     1261 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     1321 gctgtctcat cattttggca aaggccacca tgttcgtgtt tctggtgctg ctgcctctgg
     1381 tgtccagcca gtgtgtgaac ctgaccacaa gaacccagct gcctccagcc tacaccaaca
     1441 gctttaccag aggcgtgtac taccccgaca aggtgttcag atccagcgtg ctgcactcta
     1501 cccaggacct gttcctgcct ttcttcagca acgtgacctg gttccacgcc atccacgtgt
     1561 ccggcaccaa tggcaccaag agattcgaca accccgtgct gcccttcaac gacggggtgt
     1621 actttgccag caccgagaag tccaacatca tcagaggctg gatcttcggc accacactgg
     1681 acagcaagac ccagagcctg ctgatcgtga acaacgccac caacgtggtc atcaaagtgt
     1741 gcgagttcca gttctgcaac gaccccttcc tgggcgtcta ctatcacaag aacaacaaga
     1801 gctggatgga aagcgagttc cgggtgtaca gcagcgccaa caactgcacc ttcgagtacg
     1861 tgtcccagcc tttcctgatg gacctggaag gcaagcaggg caacttcaag aacctgcgcg
     1921 agttcgtgtt caagaacatc gacggctact tcaagatcta cagcaagcac acccctatca
     1981 acctcgtgcg ggatctgcct cagggcttct ctgctctgga acccctggtg gatctgccca
     2041 tcggcatcaa catcacccgg tttcagacac tgctggccct gcacagaagc tacctgacac
     2101 ctggcgatag cagcagcgga tggacagctg gtgccgccgc ttactatgtg ggctacctgc
     2161 agcctagaac ctttctgctg aagtacaacg agaacggcac catcaccgac gccgtggatt
     2221 gtgctctgga tcctctgagc gagacaaagt gcaccctgaa gtccttcacc gtggaaaagg
     2281 gcatctacca gaccagcaac ttccgggtgc agcccaccga atccatcgtg cggttcccca
     2341 atatcaccaa tctgtgcccc ttcggcgagg tgttcaatgc caccagattc gcctctgtgt
     2401 acgcctggaa ccggaagcgg atcagcaatt gcgtggccga ctactccgtg ctgtacaact
     2461 ccgccagctt cagcaccttc aagtgctacg gcgtgtcccc taccaagctg aacgacctgt
     2521 gcttcacaaa cgtgtacgcc gacagcttcg tgatccgggg agatgaagtg cggcagattg
     2581 cccctggaca gacaggcaag atcgccgact acaactacaa gctgcccgac gacttcaccg
     2641 gctgtgtgat tgcctggaac agcaacaacc tggactccaa agtcggcggc aactacaatt
     2701 acctgtaccg gctgttccgg aagtccaatc tgaagccctt cgagcgggac atctccaccg
     2761 agatctatca ggccggcagc accccttgta acggcgtgga aggcttcaac tgctacttcc
     2821 cactgcagtc ctacggcttt cagcccacaa atggcgtggg ctatcagccc tacagagtgg
     2881 tggtgctgag cttcgaactg ctgcatgccc ctgccacagt gtgcggccct aagaaaagca
     2941 ccaatctcgt gaagaacaaa tgcgtgaact tcaacttcaa cggcctgacc ggcaccggcg
     3001 tgctgacaga gagcaacaag aagttcctgc cattccagca gtttggccgg gatatcgccg
     3061 ataccacaga cgccgttaga gatccccaga cactggaaat cctggacatc accccttgca
     3121 gcttcggcgg agtgtctgtg atcacccctg gcaccaacac cagcaatcag gtggcagtgc
     3181 tgtaccagga cgtgaactgt accgaagtgc ccgtggccat tcacgccgat cagctgacac
     3241 ctacatggcg ggtgtactcc accggcagca atgtgtttca gaccagagcc ggctgtctga
     3301 tcggagccga gcacgtgaac aatagctacg agtgcgacat ccccatcggc gctggcatct
     3361 gtgccagcta ccagacacag acaaacagcc ccgcctctgt ggccagccag agcatcattg
     3421 cctacacaat gtctctgggc gccgagaaca gcgtggccta ctccaacaac tctatcgcta
     3481 tccccaccaa cttcaccatc agcgtgacca cagagatcct gcctgtgtcc atgaccaaga
     3541 ccagcgtgga ctgcaccatg tacatctgcg gcgattccac cgagtgctcc aacctgctgc
     3601 tgcagtacgg cagcttctgc acccagctga atagagccct gacagggatc gccgtggaac
     3661 aggacaagaa cacccaagag gtgttcgccc aagtgaagca gatctacaag acccctccta
     3721 tcaaggactt cggcggcttc aatttcagcc agattctgcc cgatcctagc aagcccagca
     3781 agcggagctt catcgaggac ctgctgttca acaaagtgac actggccgac gccggcttca
     3841 tcaagcagta tggcgattgt ctgggcgaca ttgccgccag ggatctgatt tgcgcccaga
     3901 agtttaacgg actgacagtg ctgcctcctc tgctgaccga tgagatgatc gcccagtaca
     3961 catctgccct gctggccggc acaatcacaa gcggctggac atttggagct ggcgccgctc
     4021 tgcagatccc ctttgctatg cagatggcct accggttcaa cggcatcgga gtgacccaga
     4081 atgtgctgta cgagaaccag aagctgatcg ccaaccagtt caacagcgcc atcggcaaga
     4141 tccaggacag cctgagcagc acagcaagcg ccctgggaaa gctgcaggac gtggtcaacc
     4201 agaatgccca ggcactgaac accctggtca agcagctgtc ctccaacttc ggcgccatca
     4261 gctctgtgct gaacgatatc ctgagcagac tggaccctcc tgaagccgag gtgcagatcg
     4321 acagactgat caccggaagg ctgcagtccc tgcagaccta cgttacccag cagctgatca
     4381 gagccgccga gattagagcc tctgccaatc tggccgccac caagatgtct gagtgtgtgc
     4441 tgggccagag caagagagtg gacttttgcg gcaagggcta ccacctgatg agcttccctc
     4501 agtctgcccc tcacggcgtg gtgtttctgc acgtgacata cgtgcccgct caagagaaga
     4561 atttcaccac cgctccagcc atctgccacg acggcaaagc ccactttcct agagaaggcg
     4621 tgttcgtgtc caacggcacc cattggttcg tgacccagcg gaacttctac gagccccaga
     4681 tcatcaccac cgacaacacc ttcgtgtctg gcaactgcga cgtcgtgatc ggcattgtga
     4741 acaataccgt gtacgaccct ctgcagcccg agctggacag cttcaaagag gaactggata
     4801 agtactttaa gaaccacaca agccccgacg tggacctggg cgatatcagc ggaatcaatg
     4861 ccagcgtcgt gaacatccag aaagagatcg accggctgaa cgaggtggcc aagaatctga
     4921 acgagagcct gatcgacctg caagaactgg ggaagtacga gcagtacatc aagtggccca
     4981 gcggccgctt ggtcccacgt ggctcacccg gatctggata catcccggag gcccctaggg
     5041 acggtcaagc ttacgtgaga aaggacggcg aatgggttct gctgtcgacc ttcttgggac
     5101 atcatcatca tcatcactaa tgaaattcga gctcgcggcc gcatcgatct taagtcgcga
     5161 ctcgagctag cagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     5221 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     5281 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     5341 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     5401 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     5461 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     5521 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     5581 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     5641 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5701 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5761 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5821 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5881 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5941 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6001 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     6061 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6121 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     6181 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6841 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7081 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac
     8161 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     8221 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     8281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     8341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     8401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     8461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     8521 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     8581 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     8641 gggcgggggg gggggggggg gsgcgnccmr gggggggggg gggggggggg gggggggggg
     8701 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     8761 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
//
