LOCUS       sequence-364434-        6786 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain from the BA.2 variant 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 6786)
  AUTHORS   Teng IT, Nazzari AF, Choe M, Liu T, Oliveira de Souza M, Petrova Y, 
            Tsybovsky Y, Wang S, Zhang B, Artamonov M, Madan B, Huang A, Lopez 
            Acevedo SN, Pan X, Ruckwardt TJ, DeKosky BJ, Mascola JR, Misasi J, 
            Sullivan NJ, Zhou T, Kwong PD
  TITLE     Molecular probes of spike ectodomain and its subdomains for 
            SARS-CoV-2 variants, Alpha through Omicron.
  JOURNAL   PLoS One. 2022 May 24;17(5):e0268767. doi: 
            10.1371/journal.pone.0268767. eCollection 2022.
  PUBMED    35609088
REFERENCE   2  (bases 1 to 6786)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2022-05-24"; volume: "17(5):e0268767. doi"; pages: " 
            10.1371/journal.pone.0268767. eCollection 2022"
COMMENT     Sequence Label: pVRC8400-SARS-CoV-2-RBD-SD1-BA.2-AVI
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        160..539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        540..743
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     694..714
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     740..764
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2673..2696
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3546..3590
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3613..3837
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4204..4221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4375..4963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4455..4474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5505..6320)
                     /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(5505..6320)
                     /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     6228..6247
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6312..6331
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6598..6616
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6654..6676)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(6776..6786,1..9)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
       61 aaataccgca tcagattggc tattggccat tgcatacgtt gtatccatat cataatatgt
      121 acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta ttgactagtt
      181 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      241 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      301 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      361 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      421 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      481 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      541 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      601 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      661 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      721 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      781 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccat cggctcgcat
      841 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
      901 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
      961 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
     1021 gctctccacg ctttgcctga ccctgcttgc tcaactctag ttaacggtgg agggcagtgt
     1081 agtctgagca gtactcgttg ctgccgcgcg cgccaccaga cataatagct gacagactaa
     1141 cagactgttc ctttccatgg gtcttttctg cagtcaccgt cgtcgacacg tgtgatcaga
     1201 tatcgcggcc gctctagacc accatgggat ggtcatgtat catccttttt ctagtagcaa
     1261 ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg acctagcgtg ttcctgtttc
     1321 cccctaagcc taaggacacc ctgatgatca gccggacccc agaggtgaca tgcgtggtgg
     1381 tggacgtgtc ccacgaggac cccgaggtga agttcaactg gtacgtggat ggcgtggagg
     1441 tgcacaatgc caagaccaag ccaagggagg agcagtacaa ctctacctat cgcgtggtga
     1501 gcgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtacaag tgtaaggtgt
     1561 ctaataaggc cctgcctgcc ccaatcgaga agaccatcag caaggcaaag ggacagccaa
     1621 gggagcctca ggtgtatacc ctgccaccca gccgcgacga gctgacaaag aaccaggtgt
     1681 ccctgtactg cctggtgaag ggcttctatc cttccgatat cgccgtggag tgggagtcta
     1741 atggccagcc agagaacaat tacaagacca cacctccagt gctggactcc gatggctctt
     1801 tctttctgta ttccaagctg accgtggata agagccggtg gcagcagggc aacgtgttta
     1861 gctgttccgt gatgcacgag gccctgcaca atcactacac acagaagtct ctgagcctgt
     1921 cccccggcaa gggaggagga ggcagcggcg gaggaggctc cggaggcggc ggctctggcg
     1981 gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt gttcctgttt ccacctaagc
     2041 ccaaagatac cctgatgatc tccaggaccc ctgaagtcac ctgcgtggtc gtggacgtgt
     2101 ctcatgagga ccccgaagtc aagttcaact ggtacgtgga cggagtcgaa gtccataacg
     2161 ccaagaccaa gcctagagaa gagcagtaca actccaccta tagagtggtg tctgtgctga
     2221 cagtgctgca tcaggattgg ctgaatggca aggaatataa gtgtaaggtg agcaataagg
     2281 ccctgcccgc ccctatcgag aagaccatct ctaaagccaa aggacagcca agggagccac
     2341 aggtgtatac actgccaccc agccgggacg agctgaccaa gaaccaggtg agcctgacat
     2401 gtctggtcaa gggcttctac cccagtgaca ttgctgtgga gtgggagtcc aatggccagc
     2461 ctgagaacaa ttataaaact accccaccag tgctggacag cgatggctcc ttctttctga
     2521 ccagcaagct gacagtggat aagtcccgct ggcagcaggg aaacgtgttt tcttgtagcg
     2581 tgatgcatga ggccctgcat aaccactaca cccagaagtc cctgtctctg agcccaggca
     2641 agggaggcag cggaggcgga ggcagcggag gcctggaggt gctgtttcag ggccctcggg
     2701 tgcagcctac agagagcatc gtgcgatttc caaacattac caacctctgc cccttcgatg
     2761 aggtgtttaa cgccacaaga tttgcctccg tttacgcctg gaatagaaag agaatcagca
     2821 attgtgtggc cgactactcc gtgctgtata actttgcccc gttcttcgcc ttcaagtgct
     2881 acggcgtttc cccaacaaag ctgaatgacc tgtgcttcac caacgtgtac gccgactcct
     2941 tcgtaattag aggcaatgag gtgagccaga tcgcaccagg ccagaccggt aacatcgctg
     3001 actacaacta taagctgcct gatgatttta caggctgcgt gatcgcctgg aactctaaca
     3061 aactggatag caaggtgggc ggcaactaca actacctgta ccggctgttt cgcaagtcta
     3121 acctgaaacc tttcgagaga gacatctcca cagagatcta ccaggccggt aataaacctt
     3181 gtaacggggt ggcaggcttc aactgttact tccctctgcg tagctacggc ttccgtccta
     3241 cctatggagt cggccaccag ccataccggg tggtcgtgct gtccttcgag ttactccacg
     3301 cccccgccac cgtctgcggt cctaagaagt ccaccaatct ggttaagaac aaatgcgtga
     3361 acttcaactt caacggcctg accgggaccg gcgtgctgac cgaaagcaac aaaaagttcc
     3421 tccccttcca gcagttcggc cgtgatatcg ctgacaccac agatgccgtc agagatccac
     3481 agaccctgga aatcctggat attacaccct gctccggagg aggcctggtg ccacagcaga
     3541 gcggaggcct gaacgacatc ttcgaggccc agaagatcga gtggcacgag ggctgatgag
     3601 gatccagatc tgctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3661 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3721 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3781 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatgggta
     3841 cccaggtgct gaagaattga cccggttcct cctgggccag aaagaagcag gcacatcccc
     3901 ttctctgtga cacaccctgt ccacgcccct ggttcttagt tccagcccca ctcataggac
     3961 actcatagct caggagggct ccgccttcaa tcccacccgc taaagtactt ggagcggtct
     4021 ctccctccct catcagccca ccaaaccaaa cctagcctcc aagagtggga agaaattaaa
     4081 gcaagatagg ctattaagtg cagagggaga gaaaatgcct ccaacatgtg aggaagtaat
     4141 gagagaaatc atagaatttt aaggccatga tttaaggcca tcatggcctt aatcttccgc
     4201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactcgggg ggggggngcg
     5221 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc
     5281 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt
     5341 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat
     5401 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc
     5461 agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     5521 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     5581 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     5641 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     5701 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     5761 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     5821 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     5881 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     5941 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     6001 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     6061 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     6121 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     6181 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     6241 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt
     6301 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     6361 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     6421 acgtggcttt ccccccnccc ccattattga agcatttatc agggttattg tctcatgagc
     6481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6541 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6601 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6661 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6721 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6781 tcagag
//
