LOCUS       sequence-314491-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-L455RA475RG502R 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 6557)
  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 6557)
  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-L455RA475RG502R-AVI
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        285..664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        665..868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     819..839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..889
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2798..2821
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3443..3487
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3510..3734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4101..4118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4272..4860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4352..4371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5401..6216)
                     /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(5401..6216)
                     /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     6124..6143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6208..6227
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6494..6512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6550..6557,1..15))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
       61 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      121 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
      181 ggagaaaata ccgcatcaga ttggctattg gccattgcat acgttgtatc catatcataa
      241 tatgtacatt tatattggct catgtccaac attaccgcca tgttgacatt gattattgac
      301 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      661 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      721 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      781 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      841 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      901 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccatcggct
      961 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     1021 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     1081 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     1141 agccggctct ccacgctttg cctgaccctg cttgctcaac tctagttaac ggtggagggc
     1201 agtgtagtct gagcagtact cgttgctgcc gcgcgcgcca ccagacataa tagctgacag
     1261 actaacagac tgttcctttc catgggtctt ttctgcagtc accgtcgtcg acacgtgtga
     1321 tcagatatcg cggccgctct agaccaccat gggatggtca tgtatcatcc tttttctagt
     1381 agcaactgca accggtgtac attccgcacc agagctgctg ggaggaccta gcgtgttcct
     1441 gtttccccct aagcctaagg acaccctgat gatcagccgg accccagagg tgacatgcgt
     1501 ggtggtggac gtgtcccacg aggaccccga ggtgaagttc aactggtacg tggatggcgt
     1561 ggaggtgcac aatgccaaga ccaagccaag ggaggagcag tacaactcta cctatcgcgt
     1621 ggtgagcgtg ctgacagtgc tgcaccagga ctggctgaac ggcaaggagt acaagtgtaa
     1681 ggtgtctaat aaggccctgc ctgccccaat cgagaagacc atcagcaagg caaagggaca
     1741 gccaagggag cctcaggtgt ataccctgcc acccagccgc gacgagctga caaagaacca
     1801 ggtgtccctg tactgcctgg tgaagggctt ctatccttcc gatatcgccg tggagtggga
     1861 gtctaatggc cagccagaga acaattacaa gaccacacct ccagtgctgg actccgatgg
     1921 ctctttcttt ctgtattcca agctgaccgt ggataagagc cggtggcagc agggcaacgt
     1981 gtttagctgt tccgtgatgc acgaggccct gcacaatcac tacacacaga agtctctgag
     2041 cctgtccccc ggcaagggag gaggaggcag cggcggagga ggctccggag gcggcggctc
     2101 tggcggcggc ggcagcgccc ctgaactgct gggaggacca agcgtgttcc tgtttccacc
     2161 taagcccaaa gataccctga tgatctccag gacccctgaa gtcacctgcg tggtcgtgga
     2221 cgtgtctcat gaggaccccg aagtcaagtt caactggtac gtggacggag tcgaagtcca
     2281 taacgccaag accaagccta gagaagagca gtacaactcc acctatagag tggtgtctgt
     2341 gctgacagtg ctgcatcagg attggctgaa tggcaaggaa tataagtgta aggtgagcaa
     2401 taaggccctg cccgccccta tcgagaagac catctctaaa gccaaaggac agccaaggga
     2461 gccacaggtg tatacactgc cacccagccg ggacgagctg accaagaacc aggtgagcct
     2521 gacatgtctg gtcaagggct tctaccccag tgacattgct gtggagtggg agtccaatgg
     2581 ccagcctgag aacaattata aaactacccc accagtgctg gacagcgatg gctccttctt
     2641 tctgaccagc aagctgacag tggataagtc ccgctggcag cagggaaacg tgttttcttg
     2701 tagcgtgatg catgaggccc tgcataacca ctacacccag aagtccctgt ctctgagccc
     2761 aggcaaggga ggcagcggag gcggaggcag cggaggcctg gaggtgctgt ttcagggccc
     2821 tcccaatatc acaaacctgt gcccttttgg cgaggtgttc aacgcaacca ggttcgccag
     2881 cgtgtacgca tggaatagga agcgcatctc caactgcgtg gccgactatt ctgtgctgta
     2941 caacagcgcc tccttctcta cctttaagtg ctatggcgtg agccccacaa agctgaatga
     3001 cctgtgcttt accaacgtgt acgccgattc cttcgtgatc aggggcgacg aggtgcgcca
     3061 gatcgcacca ggacagacag gcaagatcgc agactacaat tataagctgc ctgacgattt
     3121 caccggctgc gtgatcgcct ggaactctaa caatctggat agcaaagtgg gcggcaacta
     3181 caattatctg taccggaggt ttagaaagtc taatctgaag ccattcgaga gggacatctc
     3241 cacagaaatc taccagcgcg gctctacccc ctgcaatggc gtggagggct ttaactgtta
     3301 tttccctctg cagagctacg gcttccagcc aacaaaccgc gtgggctatc agccctaccg
     3361 cgtggtggtg ctgtcttttg agctgctgca cgcacctgca acagtgtgcg gaggaggagg
     3421 cctggtgcca cagcagagcg gaggcctgaa cgacatcttc gaggcccaga agatcgagtg
     3481 gcacgagggc tgatgaggat ccagatctgc tgtgccttct agttgccagc catctgttgt
     3541 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3601 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3661 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3721 ggtgggctct atgggtaccc aggtgctgaa gaattgaccc ggttcctcct gggccagaaa
     3781 gaagcaggca catccccttc tctgtgacac accctgtcca cgcccctggt tcttagttcc
     3841 agccccactc ataggacact catagctcag gagggctccg ccttcaatcc cacccgctaa
     3901 agtacttgga gcggtctctc cctccctcat cagcccacca aaccaaacct agcctccaag
     3961 agtgggaaga aattaaagca agataggcta ttaagtgcag agggagagaa aatgcctcca
     4021 acatgtgagg aagtaatgag agaaatcata gaattttaag gccatgattt aaggccatca
     4081 tggccttaat cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4141 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4201 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4261 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4321 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4381 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4441 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4501 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4561 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4621 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4681 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4741 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4801 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4921 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4981 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5041 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5101 ctcggggggg ggggcgctga ggtctgcctc gtgaagaagg tgttgctgac tcataccagg
     5161 cctgaatcgc cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt
     5221 tgtaggtgga ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt
     5281 cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc
     5341 gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga
     5401 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     5461 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     5521 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     5581 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     5641 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     5701 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     5761 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     5821 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     5881 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     5941 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     6001 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     6061 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     6121 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     6181 cctcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6241 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     6301 gagattttga gacacaacgt ggctttcccc ccccccccat tattgaagca tttatcaggg
     6361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6421 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6481 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     6541 cggtgaaaac ctctgac
//
