LOCUS       sequence-314421-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain 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 6636)
  AUTHORS   Zhou T, Teng IT, Olia AS, Cerutti G, Gorman J, Nazzari A, Shi W, 
            Tsybovsky Y, Wang L, Wang S, Zhang B, Zhang Y, Katsamba PS, Petrova 
            Y, Banach BB, Fahad AS, Liu L, Lopez Acevedo SN, Madan B, Oliveira 
            de Souza M, Pan X, Wang P, Wolfe JR, Yin M, Ho DD, Phung E, DiPiazza
            A, Chang LA, Abiona OM, Corbett KS, DeKosky BJ, Graham BS, Mascola 
            JR, Misasi J, Ruckwardt T, Sullivan NJ, Shapiro L, Kwong PD
  TITLE     Structure-Based Design with Tag-Based Purification and In-Process 
            Biotinylation Enable Streamlined Development of SARS-CoV-2 Spike 
            Molecular Probes.
  JOURNAL   Cell Rep. 2020 Oct 27;33(4):108322. doi: 
            10.1016/j.celrep.2020.108322. Epub 2020 Oct 12.
  PUBMED    33091382
REFERENCE   2  (bases 1 to 6636)
  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: "Cell Rep.";
            date: "2020-10-27"; volume: "33(4):108322. doi"; pages: " 
            10.1016/j.celrep.2020.108322. Epub 2020 Oct 12"
COMMENT     Sequence Label: pVRC8400-SARS-CoV-2-RBD-SD1-AVI
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     702..722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     748..772
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2681..2704
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3404..3448
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3471..3695
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4062..4079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4233..4821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4313..4332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5363..6178)
                     /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(5363..6178)
                     /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     6086..6105
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6170..6189
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6456..6474
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6512..6534)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(6634..6636,1..17)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
       61 taaggagaaa ataccgcatc agattggcta ttggccattg catacgttgt atccatatca
      121 taatatgtac atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt
      181 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      241 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      601 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      661 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      721 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
      781 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccatcg
      841 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
      901 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
      961 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     1021 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctagtt aacggtggag
     1081 ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg ccaccagaca taatagctga
     1141 cagactaaca gactgttcct ttccatgggt cttttctgca gtcaccgtcg tcgacacgtg
     1201 tgatcagata tcgcggccgc tctagaccac catgggatgg tcatgtatca tcctttttct
     1261 agtagcaact gcaaccggtg tacattccgc accagagctg ctgggaggac ctagcgtgtt
     1321 cctgtttccc cctaagccta aggacaccct gatgatcagc cggaccccag aggtgacatg
     1381 cgtggtggtg gacgtgtccc acgaggaccc cgaggtgaag ttcaactggt acgtggatgg
     1441 cgtggaggtg cacaatgcca agaccaagcc aagggaggag cagtacaact ctacctatcg
     1501 cgtggtgagc gtgctgacag tgctgcacca ggactggctg aacggcaagg agtacaagtg
     1561 taaggtgtct aataaggccc tgcctgcccc aatcgagaag accatcagca aggcaaaggg
     1621 acagccaagg gagcctcagg tgtataccct gccacccagc cgcgacgagc tgacaaagaa
     1681 ccaggtgtcc ctgtactgcc tggtgaaggg cttctatcct tccgatatcg ccgtggagtg
     1741 ggagtctaat ggccagccag agaacaatta caagaccaca cctccagtgc tggactccga
     1801 tggctctttc tttctgtatt ccaagctgac cgtggataag agccggtggc agcagggcaa
     1861 cgtgtttagc tgttccgtga tgcacgaggc cctgcacaat cactacacac agaagtctct
     1921 gagcctgtcc cccggcaagg gaggaggagg cagcggcgga ggaggctccg gaggcggcgg
     1981 ctctggcggc ggcggcagcg cccctgaact gctgggagga ccaagcgtgt tcctgtttcc
     2041 acctaagccc aaagataccc tgatgatctc caggacccct gaagtcacct gcgtggtcgt
     2101 ggacgtgtct catgaggacc ccgaagtcaa gttcaactgg tacgtggacg gagtcgaagt
     2161 ccataacgcc aagaccaagc ctagagaaga gcagtacaac tccacctata gagtggtgtc
     2221 tgtgctgaca gtgctgcatc aggattggct gaatggcaag gaatataagt gtaaggtgag
     2281 caataaggcc ctgcccgccc ctatcgagaa gaccatctct aaagccaaag gacagccaag
     2341 ggagccacag gtgtatacac tgccacccag ccgggacgag ctgaccaaga accaggtgag
     2401 cctgacatgt ctggtcaagg gcttctaccc cagtgacatt gctgtggagt gggagtccaa
     2461 tggccagcct gagaacaatt ataaaactac cccaccagtg ctggacagcg atggctcctt
     2521 ctttctgacc agcaagctga cagtggataa gtcccgctgg cagcagggaa acgtgttttc
     2581 ttgtagcgtg atgcatgagg ccctgcataa ccactacacc cagaagtccc tgtctctgag
     2641 cccaggcaag ggaggcagcg gaggcggagg cagcggaggc ctggaggtgc tgtttcaggg
     2701 ccctagggtg cagcctaccg agtccatcgt gcgctttccc aatatcacaa acctgtgccc
     2761 ttttggcgag gtgttcaacg caaccaggtt cgccagcgtg tacgcatgga ataggaagcg
     2821 catctccaac tgcgtggccg actattctgt gctgtacaac agcgcctcct tctctacctt
     2881 taagtgctat ggcgtgagcc ccacaaagct gaatgacctg tgctttacca acgtgtacgc
     2941 cgattccttc gtgatcaggg gcgacgaggt gcgccagatc gcaccaggac agacaggcaa
     3001 gatcgcagac tacaattata agctgcctga cgatttcacc ggctgcgtga tcgcctggaa
     3061 ctctaacaat ctggatagca aagtgggcgg caactacaat tatctgtacc ggctgtttag
     3121 aaagtctaat ctgaagccat tcgagaggga catctccaca gaaatctacc aggccggctc
     3181 taccccctgc aatggcgtgg agggctttaa ctgttatttc cctctgcaga gctacggctt
     3241 ccagccaaca aacggcgtgg gctatcagcc ctaccgcgtg gtggtgctgt cttttgagct
     3301 gctgcacgca cctgcaacag tgtgcggacc aaagaagagc accaatctgg tgaagaacaa
     3361 gtgcgtgaac ttcggaggag gcctggtgcc acagcagagc ggaggcctga acgacatctt
     3421 cgaggcccag aagatcgagt ggcacgaggg ctgatgagga tccagatctg ctgtgccttc
     3481 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3541 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3601 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3661 tagcaggcat gctggggatg cggtgggctc tatgggtacc caggtgctga agaattgacc
     3721 cggttcctcc tgggccagaa agaagcaggc acatcccctt ctctgtgaca caccctgtcc
     3781 acgcccctgg ttcttagttc cagccccact cataggacac tcatagctca ggagggctcc
     3841 gccttcaatc ccacccgcta aagtacttgg agcggtctct ccctccctca tcagcccacc
     3901 aaaccaaacc tagcctccaa gagtgggaag aaattaaagc aagataggct attaagtgca
     3961 gagggagaga aaatgcctcc aacatgtgag gaagtaatga gagaaatcat agaattttaa
     4021 ggccatgatt taaggccatc atggccttaa tcttccgctt cctcgctcac tgactcgctg
     4081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5041 cgttcatcca tagttgcctg actcgggggg ggggggcgct gaggtctgcc tcgtgaagaa
     5101 ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag
     5161 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
     5221 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     5281 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt
     5341 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     5401 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     5461 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     5521 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     5581 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     5641 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     5701 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     5761 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     5821 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     5881 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     5941 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     6001 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     6061 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     6121 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     6181 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     6241 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc cccccccccc
     6301 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6361 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6421 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6481 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6541 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6601 gtgttggcgg gtgtcggggc tggcttaact atgcgg
//
