LOCUS       sequence-374987-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD 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 6828)
  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  JOURNAL   Unpublished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 6828)
  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            "Unpublished"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-NTD-BA.4/5-AVI
FEATURES             Location/Qualifiers
     source          1..6828
                     /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             3749..3793
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3816..4040
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4407..4424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4578..5166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4658..4677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5708..6523)
                     /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(5708..6523)
                     /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     6431..6450
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6515..6534
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6801..6819
                     /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 gggccctcag tgcgtgaacc tgatcacccg
     2881 aacccagagc tacaccaaca gctttacacg gggcgtgtac taccctgaca aggtgttcag
     2941 atctagcgtc ctgcacagca ctcaggacct cttcctgccg ttcttcagca acgtgacatg
     3001 gttccacgcc atcagcggca caaacggaac caagcggttt gataaccccg tcctgccatt
     3061 caatgatgga gtttacttcg ccagtaccga gaagagtaac atcatccggg gctggatctt
     3121 cggcaccacc ctggatagca aaacacagag cctcctgatc gtgaacaatg ccacgaacgt
     3181 cgtgatcaag gtgtgcgagt tccagttttg caatgatcct ttcctggatg tgtactacca
     3241 caagaacaac aagagctgga tggaaagcga gttcagagtc tacagcagcg ccaacaactg
     3301 cacattcgag tacgtctctc agccttttct gatggacctt gaggggaaac aaggcaactt
     3361 caagaacctg agagaattcg tgttcaagaa catcgacggc tacttcaaaa tctactccaa
     3421 gcacacaccc atcaacctgg gccgggacct ccctcagggc ttcagcgccc tggaacccct
     3481 ggtcgacctg cccataggca tcaacataac gcggttccaa accctgctgg ccctgcatag
     3541 atcctacctg actcctggcg acagcagcag cggatggacc gccggagctg cagcctacta
     3601 tgtgggctac ctgcaaccta gaaccttcct gctgaagtac aacgagaacg gcacaatcac
     3661 agacgccgtc gactgcgccc tggaccctct ctctgagaca aagtgcaccc tgaagtccgg
     3721 aggaggcctg gtgccacagc agagcggagg cctgaacgac atcttcgagg cccagaagat
     3781 cgagtggcac gagggctgat gaggatccag atctgctgtg ccttctagtt gccagccatc
     3841 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3901 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3961 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4021 ggatgcggtg ggctctatgg gtacccaggt gctgaagaat tgacccggtt cctcctgggc
     4081 cagaaagaag caggcacatc cccttctctg tgacacaccc tgtccacgcc cctggttctt
     4141 agttccagcc ccactcatag gacactcata gctcaggagg gctccgcctt caatcccacc
     4201 cgctaaagta cttggagcgg tctctccctc cctcatcagc ccaccaaacc aaacctagcc
     4261 tccaagagtg ggaagaaatt aaagcaagat aggctattaa gtgcagaggg agagaaaatg
     4321 cctccaacat gtgaggaagt aatgagagaa atcatagaat tttaaggcca tgatttaagg
     4381 ccatcatggc cttaatcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4741 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4981 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5221 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5281 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5341 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5401 gcctgactcg gggggggggg gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca
     5461 taccaggcct gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga
     5521 gctttgttgt aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct
     5581 gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa
     5641 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
     5701 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     5761 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     5821 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     5881 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     5941 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     6001 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     6061 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     6121 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     6181 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     6241 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     6301 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     6361 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     6421 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     6481 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     6541 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     6601 aacatcagag attttgagac acaacgtggc tttccccccc cccccattat tgaagcattt
     6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6721 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6781 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
