LOCUS       sequence-348111-        8055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient mammalian expression of codon optimized SARS-CoV-2 Spike 
            proteins (D614G, N501Y). This version gives lower expression of 
            Spike and is more convenient for generating VLPs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  AUTHORS   Syed AM, Taha TY, Tabata T, Chen IP, Ciling A, Khalid MM, Sreekumar 
            B, Chen PY, Hayashi JM, Soczek KM, Ott M, Doudna JA
  TITLE     Rapid assessment of SARS-CoV-2 evolved variants using virus-like 
            particles.
  JOURNAL   Science. 2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184.
  PUBMED    34735219
REFERENCE   2  (bases 1 to 8055)
  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: "Science. 
            2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184."
COMMENT     Sequence Label: CoV2-Spike-EF1a-D614G-N501Y
FEATURES             Location/Qualifiers
     source          1..8055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..1345
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          394..1336
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1293..1313
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    5207..5795
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5260..5280)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5678..5689)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5864..5881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5870..6094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6230..6818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6310..6329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6989..7780,7781..7849))
                     /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     7612..7631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7850..7954)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8029..8048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
       61 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      121 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtgctccgg tgcccgtcag
      181 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
      241 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
      301 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
      361 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
      421 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct
      481 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
      541 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg
      601 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
      661 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
      721 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
      781 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
      841 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
      901 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
      961 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     1021 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     1081 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     1141 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     1201 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     1261 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     1321 ttttcttcca tttcaggtgt cgtgagctag catgtttgtc ttcctggtcc tgctgcctct
     1381 ggtctcgtct cagtgcgtga acctgactac tagaacccag ctgcctcctg cctatactaa
     1441 ctccttcacc cgcggcgtgt actacccaga caaggtgttc cgcagctccg tgctgcactc
     1501 cacccaggat ctgttcctgc ccttcttcag caacgtgacc tggttccacg ccatccacgt
     1561 gagcggcacc aatggcacca agcggttcga caatcccgtg ctgccattca acgatggcgt
     1621 gtacttcgcc tccaccgaga agagcaacat catccgcggc tggatcttcg gcaccaccct
     1681 ggactccaag acccagagcc tgctgatcgt gaacaatgcc accaacgtgg tcatcaaggt
     1741 gtgcgagttc cagttctgca atgatccatt cctgggcgtg tactaccaca agaacaataa
     1801 gtcctggatg gagagcgagt tccgcgtgta cagctccgcc aacaattgca ccttcgagta
     1861 cgtgtcccag cccttcctga tggacctgga gggcaagcag ggcaatttca agaacctgcg
     1921 cgagttcgtg ttcaagaata tcgatggcta cttcaagatc tactccaagc acacccccat
     1981 caacctggtg cgcgacctgc cacagggctt cagcgccctg gagccactgg tggatctgcc
     2041 aatcggcatc aacatcacca ggttccagac cctgctggcc ctgcaccgca gctacctgac
     2101 cccaggcgac agctccagcg gatggaccgc tggagctgct gcctactacg tgggctacct
     2161 gcagccccgc accttcctgc tgaagtacaa cgagaatggc accatcaccg acgccgtgga
     2221 ttgcgccctg gatccactgt ccgagacaaa gtgcaccctg aagagcttca ccgtggagaa
     2281 gggcatctac cagacctcca atttccgcgt gcagccaacc gagagcatcg tgcgcttccc
     2341 caatatcacc aacctgtgcc cattcggcga ggtgttcaac gctaccaggt tcgccagcgt
     2401 gtacgcttgg aatcgcaagc gcatctccaa ctgcgtggcc gactacagcg tgctgtacaa
     2461 ctccgccagc ttctccacct tcaagtgcta cggcgtgtcc cccaccaagc tgaatgatct
     2521 gtgcttcacc aacgtgtacg ccgatagctt cgtgatcagg ggcgacgagg tgcgccagat
     2581 cgctccagga cagaccggca agatcgctga ctacaattac aagctgcccg acgatttcac
     2641 cggctgcgtg atcgcctgga actccaacaa tctggatagc aaagtgggcg gcaactacaa
     2701 ttacctgtac cgcctgttcc gcaagtccaa tctgaagcca ttcgagcgcg acatctccac
     2761 cgagatctac caggctggaa gcaccccatg caatggagtg gagggcttca actgctactt
     2821 ccccctgcag agctacggct tccagccaac ctacggagtg ggataccagc catacagggt
     2881 ggtggtgctg tccttcgagc tgctgcacgc tccagctacc gtgtgcggac caaagaagag
     2941 caccaatctg gtgaagaaca agtgcgtgaa cttcaatttc aacggcctga ccggaaccgg
     3001 cgtgctgacc gagtccaaca agaagttcct gccattccag cagttcggaa gggacatcgc
     3061 tgataccacc gacgccgtgc gcgacccaca gaccctggag atcctggata tcaccccatg
     3121 ctccttcggc ggcgtgagcg tgatcacccc aggaaccaat accagcaacc aggtggccgt
     3181 gctgtaccag ggcgtgaatt gcaccgaggt gccagtggct atccacgctg atcagctgac
     3241 cccaacctgg cgcgtgtaca gcaccggatc caacgtgttc cagacccgcg ccggatgcct
     3301 gatcggagct gagcacgtga acaattccta cgagtgcgac atcccaatcg gagctggaat
     3361 ctgcgccagc taccagaccc agaccaactc cccaaggagg gctcgcagcg tggccagcca
     3421 gtccatcatc gcctacacca tgtccctggg cgccgagaat agcgtggcct acagcaacaa
     3481 ttccatcgcc atcccaacca acttcaccat ctccgtgacc accgagatcc tgcccgtgtc
     3541 catgaccaag accagcgtgg actgcaccat gtacatctgc ggcgattcca ccgagtgcag
     3601 caacctgctg ctgcagtacg gcagcttctg cacccagctg aatcgcgccc tgaccggaat
     3661 cgctgtggag caggataaga acacccagga ggtgttcgcc caggtgaagc agatctacaa
     3721 gaccccccca atcaaggact tcggcggctt caatttcagc cagatcctgc ccgatccaag
     3781 caagccctcc aagcgcagct tcatcgagga cctgctgttc aacaaggtga ccctggccga
     3841 tgccggcttc atcaagcagt acggcgattg cctgggcgac atcgctgccc gcgacctgat
     3901 ctgcgcccag aagttcaatg gcctgaccgt gctgccacca ctgctgaccg atgagatgat
     3961 cgctcagtac acctccgccc tgctggccgg aaccatcacc agcggatgga ccttcggcgc
     4021 tggagccgcc ctgcagatcc ccttcgccat gcagatggcc taccgcttca acggcatcgg
     4081 cgtgacccag aatgtgctgt acgagaacca gaagctgatc gccaatcagt tcaactccgc
     4141 catcggcaag atccaggact ccctgtccag caccgccagc gccctgggca agctgcagga
     4201 tgtggtgaat cagaacgccc aggccctgaa taccctggtg aagcagctgt ccagcaactt
     4261 cggcgccatc tccagcgtgc tgaatgatat cctgagccgc ctggacaagg tggaggctga
     4321 ggtgcagatc gataggctga tcaccggccg cctgcagtcc ctgcagacct acgtgaccca
     4381 gcagctgatc agggctgctg agatcagggc cagcgccaat ctggctgcta ccaagatgtc
     4441 cgagtgcgtg ctgggacaga gcaagagggt ggacttctgc ggcaagggct accacctgat
     4501 gtccttccca cagagcgccc cacacggagt ggtgttcctg cacgtgacct acgtgccagc
     4561 ccaggagaag aacttcacca ccgctccagc tatctgccac gatggcaagg ctcacttccc
     4621 acgcgagggc gtgttcgtgt ccaacggcac ccactggttc gtgacccagc gcaatttcta
     4681 cgagccccag atcatcacca ccgacaatac cttcgtgagc ggcaactgcg acgtggtcat
     4741 cggaatcgtg aacaataccg tgtacgatcc cctgcagcca gagctggact ccttcaagga
     4801 ggagctggat aagtacttca agaatcacac cagccccgac gtggatctgg gcgacatctc
     4861 cggcatcaat gccagcgtgg tgaacatcca gaaggagatc gaccgcctga acgaggtggc
     4921 caagaatctg aacgagtccc tgatcgatct gcaggagctg ggcaagtacg agcagtacat
     4981 caagtggcca tggtacatct ggctgggctt catcgccggc ctgatcgcca tcgtgatggt
     5041 gaccatcatg ctgtgctgca tgacctcctg ctgcagctgc ctgaagggct gctgctcctg
     5101 cggcagctgc tgcaagttcg atgaggacga tagcgagccc gtgctgaagg gcgtcaaact
     5161 gcactataca tagctcgaca tataaggatc cactcgacga ctcgacaatc aacctctgga
     5221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5341 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5401 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5461 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5521 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5581 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5641 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5701 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg catcttcgcc ttcgccctca
     5761 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattcga gctcggtacc
     5821 ttgattctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     5881 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5941 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     6001 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     6061 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagtcaa
     6121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6421 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6661 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7981 acctccacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     8041 tgatgccgca tagtt
//
