LOCUS       sequence-374985-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD protein from the BA.4 and BA.5 variants 
            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)
  TITLE     Molecular probes of spike ectodomain and its subdomains for 
            SARS-CoV-2 BA.4 and BA.5 variants
  JOURNAL   Unpublished
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: 
            "Unpublished"
COMMENT     Sequence Label: pVRC8400-SARS-CoV-2-RBD-BA.4/5-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        321..700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        701..904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     855..875
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     901..925
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2834..2857
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3479..3523
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3546..3770
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4137..4154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4308..4896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4388..4407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5438..6253)
                     /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(5438..6253)
                     /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     6161..6180
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6245..6264
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6531..6549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcagattgg
      241 ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
      301 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      721 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      781 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      841 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      901 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
      961 tagaagacac cgggaccgat ccagcctcca tcggctcgca tctctccttc acgcgcccgc
     1021 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     1081 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     1141 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     1201 accctgcttg ctcaactcta gttaacggtg gagggcagtg tagtctgagc agtactcgtt
     1261 gctgccgcgc gcgccaccag acataatagc tgacagacta acagactgtt cctttccatg
     1321 ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag atatcgcggc cgctctagac
     1381 caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg gtgtacattc
     1441 cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc ctaaggacac
     1501 cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt cccacgagga
     1561 ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg ccaagaccaa
     1621 gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga cagtgctgca
     1681 ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg ccctgcctgc
     1741 cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc aggtgtatac
     1801 cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact gcctggtgaa
     1861 gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc cagagaacaa
     1921 ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt attccaagct
     1981 gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg tgatgcacga
     2041 ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca agggaggagg
     2101 aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca gcgcccctga
     2161 actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata ccctgatgat
     2221 ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg accccgaagt
     2281 caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca agcctagaga
     2341 agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc atcaggattg
     2401 gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg cccctatcga
     2461 gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata cactgccacc
     2521 cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca agggcttcta
     2581 ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca attataaaac
     2641 taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc tgacagtgga
     2701 taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg aggccctgca
     2761 taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca gcggaggcgg
     2821 aggcagcgga ggcctggagg tgctgtttca gggccctcca aacattacca acctctgccc
     2881 cttcgacgag gtgtttaacg ccacaagatt tgcctccgtt tacgcctgga atagaaagag
     2941 aatcagcaat tgtgtggccg actactccgt gctgtataac ttcgcccctt tcttcgcctt
     3001 caagtgctac ggcgtttccc caacaaagct gaatgacctg tgcttcacca acgtgtacgc
     3061 cgactccttc gtaattagag gcaatgaggt gtcccagatc gcaccaggcc agaccggtaa
     3121 catcgctgac tacaactata agctgcctga tgattttaca ggctgcgtga tcgcctggaa
     3181 ctccaacaag ctggatagca aggtgggcgg caactacaac tacagatacc ggctgtttcg
     3241 caagtctaac ctgaaacctt tcgagagaga catctccaca gagatctacc aggccggtaa
     3301 caagccttgt aacggggtgg ccggcgtgaa ctgttacttc cctctgcaaa gctacggctt
     3361 ccggcctacc tatggagtcg gccaccagcc ataccgggtg gtcgtgctgt ccttcgagtt
     3421 actccacgcc cccgccaccg tctgcggtgg aggaggcctg gtgccacagc agagcggagg
     3481 cctgaacgac atcttcgagg cccagaagat cgagtggcac gagggctgat gaggatccag
     3541 atctgctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3601 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3661 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3721 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gtacccaggt
     3781 gctgaagaat tgacccggtt cctcctgggc cagaaagaag caggcacatc cccttctctg
     3841 tgacacaccc tgtccacgcc cctggttctt agttccagcc ccactcatag gacactcata
     3901 gctcaggagg gctccgcctt caatcccacc cgctaaagta cttggagcgg tctctccctc
     3961 cctcatcagc ccaccaaacc aaacctagcc tccaagagtg ggaagaaatt aaagcaagat
     4021 aggctattaa gtgcagaggg agagaaaatg cctccaacat gtgaggaagt aatgagagaa
     4081 atcatagaat tttaaggcca tgatttaagg ccatcatggc cttaatcttc cgcttcctcg
     4141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4741 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4861 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4981 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5041 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5101 gcgatctgtc tatttcgttc atccatagtt gcctgactcg gggggggggg gcgctgaggt
     5161 ctgcctcgtg aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc
     5221 cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt
     5281 ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc
     5341 ttcaactcag caaaagttcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa
     5401 tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca
     5461 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     5521 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     5581 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     5641 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     5701 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     5761 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     5821 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     5881 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     5941 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     6001 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     6061 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     6121 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     6181 catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt
     6241 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     6301 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
     6361 tttccccccc cccccattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6481 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6541 tcacgaggcc ctttcgtc
//
