LOCUS       sequence-310799-        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"
     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             3557..3601
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3624..3848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4215..4232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4386..4974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4466..4485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5516..6331)
                     /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(5516..6331)
                     /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     6239..6258
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6323..6342
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6609..6627
                     /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 tgctgttcca gggccccagg gtgcagccta ccgagtccat
     2881 cgtgcgcttt cccaatatca caaacctgtg cccttttggc gaggtgttca acgcaaccag
     2941 gttcgccagc gtgtacgcat ggaataggaa gcgcatctcc aactgcgtgg ccgactattc
     3001 tgtgctgtac aacagcgcct ccttctctac ctttaagtgc tatggcgtga gccccacaaa
     3061 gctgaatgac ctgtgcttta ccaacgtgta cgccgattcc ttcgtgatca ggggcgacga
     3121 ggtgcgccag atcgcaccag gacagacagg caagatcgca gactacaatt ataagctgcc
     3181 tgacgatttc accggctgcg tgatcgcctg gaactctaac aatctggata gcaaagtggg
     3241 cggcaactac aattatctgt accggctgtt tagaaagtct aatctgaagc cattcgagag
     3301 ggacatctcc acagaaatct accaggccgg ctctaccccc tgcaatggcg tggagggctt
     3361 taactgttat ttccctctgc agagctacgg cttccagcca acaaacggcg tgggctatca
     3421 gccctaccgc gtggtggtgc tgtcttttga gctgctgcac gcacctgcaa cagtgtgcgg
     3481 accaaagaag agcaccaatc tggtgaagaa caagtgcgtg aacttcggag gaggcctggt
     3541 gccacagcag agcggaggcc tgaacgacat cttcgaggcc cagaagatcg agtggcacga
     3601 gggctgatga ggatccagat ctgctgtgcc ttctagttgc cagccatctg ttgtttgccc
     3661 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3721 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3781 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3841 ctctatgggt acccaggtgc tgaagaattg acccggttcc tcctgggcca gaaagaagca
     3901 ggcacatccc cttctctgtg acacaccctg tccacgcccc tggttcttag ttccagcccc
     3961 actcatagga cactcatagc tcaggagggc tccgccttca atcccacccg ctaaagtact
     4021 tggagcggtc tctccctccc tcatcagccc accaaaccaa acctagcctc caagagtggg
     4081 aagaaattaa agcaagatag gctattaagt gcagagggag agaaaatgcc tccaacatgt
     4141 gaggaagtaa tgagagaaat catagaattt taaggccatg atttaaggcc atcatggcct
     4201 taatcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcggg
     5221 gggggggggc gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga
     5281 atcgccccat catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag
     5341 gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
     5401 agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt
     5461 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     5521 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     5581 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5641 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5701 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5761 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     5821 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     5881 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     5941 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     6001 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     6061 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     6121 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     6181 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     6241 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     6301 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     6361 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     6421 tttgagacac aacgtggctt tccccccccc cccattattg aagcatttat cagggttatt
     6481 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6541 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6601 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
