LOCUS       sequence-347380-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing L452R mutation 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 6558)
  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 6558)
  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-L452R-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..59
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(97..119)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     219..238
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        389..768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        769..972
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     923..943
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     969..993
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2902..2925
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3547..3591
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3614..3838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4205..4222)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4376..4964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4456..4475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5506..6321)
                     /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(5506..6321)
                     /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     6229..6248
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6313..6332
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
       61 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      121 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      181 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      241 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      301 cagattggct attggccatt gcatacgttg tatccatatc ataatatgta catttatatt
      361 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa
      421 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      481 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      541 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      601 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      661 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      721 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      781 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      841 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      901 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      961 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     1021 gacctccata gaagacaccg ggaccgatcc agcctccatc ggctcgcatc tctccttcac
     1081 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     1141 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     1201 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     1261 tttgcctgac cctgcttgct caactctagt taacggtgga gggcagtgta gtctgagcag
     1321 tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc
     1381 tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg
     1441 ctctagacca ccatgggatg gtcatgtatc atcctttttc tagtagcaac tgcaaccggt
     1501 gtacattccg caccagagct gctgggagga cctagcgtgt tcctgtttcc ccctaagcct
     1561 aaggacaccc tgatgatcag ccggacccca gaggtgacat gcgtggtggt ggacgtgtcc
     1621 cacgaggacc ccgaggtgaa gttcaactgg tacgtggatg gcgtggaggt gcacaatgcc
     1681 aagaccaagc caagggagga gcagtacaac tctacctatc gcgtggtgag cgtgctgaca
     1741 gtgctgcacc aggactggct gaacggcaag gagtacaagt gtaaggtgtc taataaggcc
     1801 ctgcctgccc caatcgagaa gaccatcagc aaggcaaagg gacagccaag ggagcctcag
     1861 gtgtataccc tgccacccag ccgcgacgag ctgacaaaga accaggtgtc cctgtactgc
     1921 ctggtgaagg gcttctatcc ttccgatatc gccgtggagt gggagtctaa tggccagcca
     1981 gagaacaatt acaagaccac acctccagtg ctggactccg atggctcttt ctttctgtat
     2041 tccaagctga ccgtggataa gagccggtgg cagcagggca acgtgtttag ctgttccgtg
     2101 atgcacgagg ccctgcacaa tcactacaca cagaagtctc tgagcctgtc ccccggcaag
     2161 ggaggaggag gcagcggcgg aggaggctcc ggaggcggcg gctctggcgg cggcggcagc
     2221 gcccctgaac tgctgggagg accaagcgtg ttcctgtttc cacctaagcc caaagatacc
     2281 ctgatgatct ccaggacccc tgaagtcacc tgcgtggtcg tggacgtgtc tcatgaggac
     2341 cccgaagtca agttcaactg gtacgtggac ggagtcgaag tccataacgc caagaccaag
     2401 cctagagaag agcagtacaa ctccacctat agagtggtgt ctgtgctgac agtgctgcat
     2461 caggattggc tgaatggcaa ggaatataag tgtaaggtga gcaataaggc cctgcccgcc
     2521 cctatcgaga agaccatctc taaagccaaa ggacagccaa gggagccaca ggtgtataca
     2581 ctgccaccca gccgggacga gctgaccaag aaccaggtga gcctgacatg tctggtcaag
     2641 ggcttctacc ccagtgacat tgctgtggag tgggagtcca atggccagcc tgagaacaat
     2701 tataaaacta ccccaccagt gctggacagc gatggctcct tctttctgac cagcaagctg
     2761 acagtggata agtcccgctg gcagcaggga aacgtgtttt cttgtagcgt gatgcatgag
     2821 gccctgcata accactacac ccagaagtcc ctgtctctga gcccaggcaa gggaggcagc
     2881 ggaggcggag gcagcggagg cctggaggtg ctgtttcagg gccctccaaa cattaccaac
     2941 ctctgcccct tcggcgaggt gtttaacgcc acaagatttg cctccgttta cgcctggaat
     3001 agaaagagaa tcagcaattg tgtggccgac tactccgtgc tgtataacag cgcctctttc
     3061 agcaccttca agtgctacgg cgtttcccca acaaagctga atgacctgtg cttcaccaac
     3121 gtgtacgccg actccttcgt aattagaggc gatgaggtgc ggcagatcgc accaggccag
     3181 accggtaaga tcgctgacta caactataag ctgcctgatg attttacagg ctgcgtgatc
     3241 gcctggaact ctaacaacct ggatagcaag gtgggcggca actacaacta ccggtaccgg
     3301 ctgtttcgca agtctaacct gaaacctttc gagagagaca tctccacaga gatctaccag
     3361 gccggttcta caccttgtaa cggggtggaa ggcttcaact gttacttccc tctgcaaagc
     3421 tacggcttcc agcctaccaa tggagtcggc taccagccat accgggtggt cgtgctgtcc
     3481 ttcgagttac tccacgcccc cgccaccgtc tgcggtggag gaggcctggt gccacagcag
     3541 agcggaggcc tgaacgacat cttcgaggcc cagaagatcg agtggcacga gggctgatga
     3601 ggatccagat ctgctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3661 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3721 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3781 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatgggt
     3841 acccaggtgc tgaagaattg acccggttcc tcctgggcca gaaagaagca ggcacatccc
     3901 cttctctgtg acacaccctg tccacgcccc tggttcttag ttccagcccc actcatagga
     3961 cactcatagc tcaggagggc tccgccttca atcccacccg ctaaagtact tggagcggtc
     4021 tctccctccc tcatcagccc accaaaccaa acctagcctc caagagtggg aagaaattaa
     4081 agcaagatag gctattaagt gcagagggag agaaaatgcc tccaacatgt gaggaagtaa
     4141 tgagagaaat catagaattt taaggccatg atttaaggcc atcatggcct taatcttccg
     4201 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4261 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4321 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4381 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4441 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4501 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4561 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4621 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4681 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4741 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4801 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4861 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4921 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4981 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5041 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5101 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcggg gggggggggc
     5221 gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat
     5281 catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt
     5341 tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga
     5401 tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt
     5461 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     5521 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     5581 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     5641 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     5701 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     5761 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     5821 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     5881 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     5941 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     6001 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     6061 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     6121 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     6181 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     6241 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt
     6301 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     6361 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     6421 aacgtggctt tccccccccc cccattattg aagcatttat cagggttatt gtctcatgag
     6481 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6541 ccgaaaagtg ccacctga
//
