LOCUS       sequence-314861-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein 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 9648)
  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 9648)
  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-S2P-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /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             6569..6613
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6636..6860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7227..7244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7398..7986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7478..7497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8528..9343)
                     /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(8528..9343)
                     /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     9251..9270
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9335..9354
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9621..9639
                     /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 tgaccacccg
     2881 aacccagctc ccaccagcct acaccaacag ctttacacgg ggcgtgtact accctgacaa
     2941 ggtgttcaga tctagcgtcc tgcacagcac tcaggacctc ttcctgccgt tcttcagcaa
     3001 cgtgacatgg ttccacgcca tccacgtgag cggcacaaac ggaaccaagc ggtttgataa
     3061 ccccgtcctg ccattcaatg atggagttta cttcgccagt accgagaaga gtaacatcat
     3121 ccggggctgg atcttcggca ccaccctgga tagcaaaaca cagagcctcc tgatcgtgaa
     3181 caatgccacg aacgtcgtga tcaaggtgtg cgagttccag ttttgcaatg atcctttcct
     3241 gggtgtgtac taccacaaga acaacaagag ctggatggaa agcgagttca gagtctacag
     3301 cagcgccaac aactgcacat tcgagtacgt ctctcagcct tttctgatgg accttgaggg
     3361 gaaacaaggc aacttcaaga acctgagaga attcgtgttc aagaacatcg acggctactt
     3421 caaaatctac tccaagcaca cacccatcaa cctggtccgg gacctccctc agggcttcag
     3481 cgccctggaa cccctggtcg acctgcccat aggcatcaac ataacgcggt tccaaaccct
     3541 gctggccctg catagatcct acctgactcc tggcgacagc agcagcggat ggaccgccgg
     3601 agctgcagcc tactatgtgg gctacctgca acctagaacc ttcctgctga agtacaacga
     3661 gaacggcaca atcacagacg ccgtcgactg cgccctggac cctctctctg agacaaagtg
     3721 caccctgaag tccttcaccg tggaaaaggg catctaccag accagcaact tccgggtgca
     3781 gcctacagag agcatcgtgc gatttccaaa cattaccaac ctctgcccct tcggcgaggt
     3841 gtttaacgcc acaagatttg cctccgttta cgcctggaat agaaagagaa tcagcaattg
     3901 tgtggccgac tactccgtgc tgtataacag cgcctctttc agcaccttca agtgctacgg
     3961 cgtttcccca acaaagctga atgacctgtg cttcaccaac gtgtacgccg actccttcgt
     4021 aattagaggc gatgaggtgc ggcagatcgc accaggccag accggtaaga tcgctgacta
     4081 caactataag ctgcctgatg attttacagg ctgcgtgatc gcctggaact ctaacaacct
     4141 ggatagcaag gtgggcggca actacaacta cctgtaccgg ctgtttcgca agtctaacct
     4201 gaaacctttc gagagagaca tctccacaga gatctaccag gccggttcta caccttgtaa
     4261 cggggtggaa ggcttcaact gttacttccc tctgcaaagc tacggcttcc agcctaccaa
     4321 tggagtcggc taccagccat accgggtggt cgtgctgtcc ttcgagttac tccacgcccc
     4381 cgccaccgtc tgcggtccta agaagtccac caatctggtt aagaacaaat gcgtgaactt
     4441 caacttcaac ggcctgaccg ggaccggcgt gctgaccgaa agcaacaaaa agttcctccc
     4501 cttccagcag ttcggccgtg atatcgctga caccacagat gccgtcagag atccacagac
     4561 cctggaaatc ctggatatta caccctgctc cttcggagga gtttctgtga tcacccccgg
     4621 gaccaatacc agcaaccagg tggctgtgct gtaccaagat gttaactgca ccgaggttcc
     4681 tgtggccatc cacgccgatc agctgacacc tacttggaga gtgtactcca ctggctccaa
     4741 tgtgttccag accagggccg gatgtctgat cggcgccgag cacgtgaata acagttacga
     4801 gtgcgacatc cctatcggcg ccggcatctg tgccagctac cagacccaga caaacagccc
     4861 tgggtctgct tcctctgtag ctagccagag catcatcgcc tacaccatga gcctgggcgc
     4921 agagaacagc gtggcctatt ccaacaactc tatcgccatt cccaccaact ttacaattag
     4981 cgtcacaaca gagatcctgc ccgtgagcat gaccaagacc agcgtggact gtacaatgta
     5041 catctgtggc gacagcactg aatgcagcaa cctgctgctg caatacggct ccttttgcac
     5101 ccaactgaac cgggcgctga ccggaatcgc cgtggaacag gacaaaaata cccaggaggt
     5161 gttcgcccaa gtgaagcaga tctacaagac cccacctatc aaggacttcg gcggctttaa
     5221 ctttagccag attctccctg atccttctaa gcctagcaag cggagcttta tcgaggatct
     5281 gctgttcaac aaggtcaccc tggccgatgc cggctttatc aaacagtatg gcgattgcct
     5341 gggcgacata gccgccagag atctgatctg cgcccagaaa ttcaacggcc tgacagttct
     5401 cccacctctg ctgaccgacg agatgatcgc tcagtacacc tctgccctgc tggctggcac
     5461 catcacatct gggtggacat ttggcgccgg cgccgccctg cagatcccct ttgccatgca
     5521 gatggcctat agattcaacg gaatcggcgt gacccagaac gtgctgtatg aaaaccagaa
     5581 gctgatcgct aaccagttca attctgccat cggcaagatc caggactccc tctcctctac
     5641 cgccagcgcc ctgggcaaac tgcaggacgt ggtgaatcag aacgcccaag ccctgaacac
     5701 cctggtgaag cagctcagca gcaattttgg cgccatcagc tctgtgctga acgatatcct
     5761 gtctagactg gaccctccag aagccgaagt ccagatcgat agactgatca caggcagact
     5821 gcagtccctg caaacctacg tgacccaaca gctgatcagg gccgctgaaa taagagccag
     5881 cgccaatctc gccgctacca agatgtccga gtgtgtgctg ggacagtcta aacgcgttga
     5941 cttctgcggc aaaggctatc acctgatgag cttcccccag agcgcgccgc acggcgtggt
     6001 gttcctgcat gtgacatacg tgcctgccca agagaagaat ttcacaaccg cccctgccat
     6061 ctgccacgac ggcaaggccc acttcccaag agagggcgtt ttcgtttcca atggcacaca
     6121 ctggttcgtg acacaaagaa acttctacga accccagatt atcaccaccg acaacacctt
     6181 cgtgagtggc aattgtgacg tggtcatcgg aatcgtgaac aacacagtgt acgaccctct
     6241 gcaacctgag ctggactctt ttaaggaaga gctggacaag tactttaaaa accacaccag
     6301 ccccgatgtg gacctgggcg acatcagtgg cattaacgcc agcgtggtga acatccaaaa
     6361 ggaaatcgac agactgaacg aggtggccaa gaacctgaac gagtccctga tcgacctgca
     6421 ggagctcggc aaatacgagc agggatctgg atacatcccc gaggccccca gagatggcca
     6481 ggcctacgtg cggaaggacg gcgagtgggt actgctgagc acattcctgg gcagatccgg
     6541 aggaggcctg gtgccacagc agagcggagg cctgaacgac atcttcgagg cccagaagat
     6601 cgagtggcac gagggctgat gaggatccag atctgctgtg ccttctagtt gccagccatc
     6661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     6841 ggatgcggtg ggctctatgg gtacccaggt gctgaagaat tgacccggtt cctcctgggc
     6901 cagaaagaag caggcacatc cccttctctg tgacacaccc tgtccacgcc cctggttctt
     6961 agttccagcc ccactcatag gacactcata gctcaggagg gctccgcctt caatcccacc
     7021 cgctaaagta cttggagcgg tctctccctc cctcatcagc ccaccaaacc aaacctagcc
     7081 tccaagagtg ggaagaaatt aaagcaagat aggctattaa gtgcagaggg agagaaaatg
     7141 cctccaacat gtgaggaagt aatgagagaa atcatagaat tttaaggcca tgatttaagg
     7201 ccatcatggc cttaatcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7561 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7801 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8041 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8101 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8161 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8221 gcctgactcg gggggggggg gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca
     8281 taccaggcct gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga
     8341 gctttgttgt aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct
     8401 gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa
     8461 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
     8521 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     8581 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     8641 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     8701 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     8761 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     8821 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     8881 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     8941 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     9001 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     9061 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     9121 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     9181 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     9241 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     9301 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     9361 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     9421 aacatcagag attttgagac acaacgtggc tttccccccc cccccattat tgaagcattt
     9481 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9541 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     9601 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
