LOCUS       sequence-347378-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain containing K417N-L452R-T478K 
            mutations with single chain Fc, HRV3C protease cleavage site, and 
            AVI tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6786)
  AUTHORS   Teng IT, Nazzari AF, Choe M, Liu T, Oliveira de Souza M, Petrova Y, 
            Tsybovsky Y, Wang S, Zhang B, Artamonov M, Madan B, Huang A, Lopez 
            Acevedo SN, Pan X, Ruckwardt TJ, DeKosky BJ, Mascola JR, Misasi J, 
            Sullivan NJ, Zhou T, Kwong PD
  TITLE     Molecular probes of spike ectodomain and its subdomains for 
            SARS-CoV-2 variants, Alpha through Omicron.
  JOURNAL   PLoS One. 2022 May 24;17(5):e0268767. doi: 
            10.1371/journal.pone.0268767. eCollection 2022.
  PUBMED    35609088
REFERENCE   2  (bases 1 to 6786)
  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: "PLoS One.";
            date: "2022-05-24"; volume: "17(5):e0268767. doi"; pages: " 
            10.1371/journal.pone.0268767. eCollection 2022"
COMMENT     Sequence Label: pVRC8400-SARS-CoV-2-RBD-SD1-K417N-L452R-T478K-...
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(174..989)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(174..989)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     897..916
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     981..1000
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1267..1285
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1323..1345)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1445..1464
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1615..1994
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1995..2198
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2149..2169
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2195..2219
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4128..4151
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5001..5045
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5068..5292
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5659..5676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5830..6418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5910..5929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
       61 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc
      121 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
      181 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
      241 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
      301 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
      361 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
      421 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
      481 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
      541 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
      601 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
      661 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
      721 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
      781 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
      841 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
      901 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
      961 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     1021 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     1081 tgagacacaa cgtggctttc cccccccccc cattattgaa gcatttatca gggttattgt
     1141 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1201 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     1261 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     1321 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     1381 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     1441 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     1501 agatgcgtaa ggagaaaata ccgcatcaga ttggctattg gccattgcat acgttgtatc
     1561 catatcataa tatgtacatt tatattggct catgtccaac attaccgcca tgttgacatt
     1621 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     1681 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     1741 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     1801 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1861 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1921 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1981 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2041 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2101 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2161 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     2221 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     2281 tccatcggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     2341 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     2401 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     2461 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctagttaac
     2521 ggtggagggc agtgtagtct gagcagtact cgttgctgcc gcgcgcgcca ccagacataa
     2581 tagctgacag actaacagac tgttcctttc catgggtctt ttctgcagtc accgtcgtcg
     2641 acacgtgtga tcagatatcg cggccgctct agaccaccat gggatggtca tgtatcatcc
     2701 tttttctagt agcaactgca accggtgtac attccgcacc agagctgctg ggaggaccta
     2761 gcgtgttcct gtttccccct aagcctaagg acaccctgat gatcagccgg accccagagg
     2821 tgacatgcgt ggtggtggac gtgtcccacg aggaccccga ggtgaagttc aactggtacg
     2881 tggatggcgt ggaggtgcac aatgccaaga ccaagccaag ggaggagcag tacaactcta
     2941 cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga ctggctgaac ggcaaggagt
     3001 acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat cgagaagacc atcagcaagg
     3061 caaagggaca gccaagggag cctcaggtgt ataccctgcc acccagccgc gacgagctga
     3121 caaagaacca ggtgtccctg tactgcctgg tgaagggctt ctatccttcc gatatcgccg
     3181 tggagtggga gtctaatggc cagccagaga acaattacaa gaccacacct ccagtgctgg
     3241 actccgatgg ctctttcttt ctgtattcca agctgaccgt ggataagagc cggtggcagc
     3301 agggcaacgt gtttagctgt tccgtgatgc acgaggccct gcacaatcac tacacacaga
     3361 agtctctgag cctgtccccc ggcaagggag gaggaggcag cggcggagga ggctccggag
     3421 gcggcggctc tggcggcggc ggcagcgccc ctgaactgct gggaggacca agcgtgttcc
     3481 tgtttccacc taagcccaaa gataccctga tgatctccag gacccctgaa gtcacctgcg
     3541 tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt caactggtac gtggacggag
     3601 tcgaagtcca taacgccaag accaagccta gagaagagca gtacaactcc acctatagag
     3661 tggtgtctgt gctgacagtg ctgcatcagg attggctgaa tggcaaggaa tataagtgta
     3721 aggtgagcaa taaggccctg cccgccccta tcgagaagac catctctaaa gccaaaggac
     3781 agccaaggga gccacaggtg tatacactgc cacccagccg ggacgagctg accaagaacc
     3841 aggtgagcct gacatgtctg gtcaagggct tctaccccag tgacattgct gtggagtggg
     3901 agtccaatgg ccagcctgag aacaattata aaactacccc accagtgctg gacagcgatg
     3961 gctccttctt tctgaccagc aagctgacag tggataagtc ccgctggcag cagggaaacg
     4021 tgttttcttg tagcgtgatg catgaggccc tgcataacca ctacacccag aagtccctgt
     4081 ctctgagccc aggcaaggga ggcagcggag gcggaggcag cggaggcctg gaggtgctgt
     4141 ttcagggccc tcgggtgcag cctacagaga gcatcgtgcg atttccaaac attaccaacc
     4201 tctgcccctt cggcgaggtg tttaacgcca caagatttgc ctccgtttac gcctggaata
     4261 gaaagagaat cagcaattgt gtggccgact actccgtgct gtataacagc gcctctttca
     4321 gcaccttcaa gtgctacggc gtttccccaa caaagctgaa tgacctgtgc ttcaccaacg
     4381 tgtacgccga ctccttcgta attagaggcg atgaggtgcg gcagatcgca ccaggccaga
     4441 ccggtaacat cgctgactac aactataagc tgcctgatga ttttacaggc tgcgtgatcg
     4501 cctggaactc taacaacctg gatagcaagg tgggcggcaa ctacaactac agataccggc
     4561 tgtttcgcaa gtctaacctg aaacctttcg agagagacat ctccacagag atctaccagg
     4621 ccggttctaa gccttgtaac ggggtggaag gcttcaactg ttacttccct ctgcaaagct
     4681 acggcttcca gcctaccaat ggagtcggct accagccata ccgggtggtc gtgctgtcct
     4741 tcgagttact ccacgccccc gccaccgtct gcggtcctaa gaagtccacc aatctggtta
     4801 agaacaaatg cgtgaacttc aacttcaacg gcctgactgg gaccggcgtg ctgaccgaaa
     4861 gcaacaaaaa gttcctcccc ttccagcagt tcggccgtga tatcgctgac accacagatg
     4921 ccgtcagaga tccacagacc ctggaaatcc tggatattac accctgctcc ggaggaggcc
     4981 tggtgccaca gcagagcgga ggcctgaacg acatcttcga ggcccagaag atcgagtggc
     5041 acgagggctg atgaggatcc agatctgctg tgccttctag ttgccagcca tctgttgttt
     5101 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5161 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5221 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     5281 tgggctctat gggtacccag gtgctgaaga attgacccgg ttcctcctgg gccagaaaga
     5341 agcaggcaca tccccttctc tgtgacacac cctgtccacg cccctggttc ttagttccag
     5401 ccccactcat aggacactca tagctcagga gggctccgcc ttcaatccca cccgctaaag
     5461 tacttggagc ggtctctccc tccctcatca gcccaccaaa ccaaacctag cctccaagag
     5521 tgggaagaaa ttaaagcaag ataggctatt aagtgcagag ggagagaaaa tgcctccaac
     5581 atgtgaggaa gtaatgagag aaatcataga attttaaggc catgatttaa ggccatcatg
     5641 gccttaatct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5701 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5761 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5821 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5881 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5941 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6001 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6061 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6241 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6481 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6541 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6601 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6661 cggggggggg gggcgctgag gtctgcctcg tgaagaaggt gttgctgact cataccaggc
     6721 ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga gagctttgtt
     6781 gtaggt
//
