LOCUS       sequence-347142-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.617.1 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 9648)
  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 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: "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-S2P-B.1.617.1-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     722..741
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     806..825
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1092..1110
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1148..1170)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1270..1289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1440..1819
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1820..2023
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1974..1994
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2020..2044
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3953..3976
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             7688..7732
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    7755..7979
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8346..8363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8517..9105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8597..8616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9647..9648,1..814))
                     /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(join(9647..9648,1..814))
                     /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"
ORIGIN
        1 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
       61 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
      121 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
      181 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
      241 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
      301 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
      361 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
      421 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
      481 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
      541 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
      601 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
      661 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
      721 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
      781 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
      841 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
      901 gattttgaga cacaacgtgg ctttcccccc ccccccatta ttgaagcatt tatcagggtt
      961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1021 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1081 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     1141 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     1201 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     1261 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     1321 cgcacagatg cgtaaggaga aaataccgca tcagattggc tattggccat tgcatacgtt
     1381 gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg
     1441 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1501 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1561 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1621 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1681 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1741 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1801 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     1861 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     1921 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     1981 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     2041 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     2101 cagcctccat cggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     2161 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     2221 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     2281 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctag
     2341 ttaacggtgg agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga
     2401 cataatagct gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt
     2461 cgtcgacacg tgtgatcaga tatcgcggcc gctctagacc accatgggat ggtcatgtat
     2521 catccttttt ctagtagcaa ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg
     2581 acctagcgtg ttcctgtttc cccctaagcc taaggacacc ctgatgatca gccggacccc
     2641 agaggtgaca tgcgtggtgg tggacgtgtc ccacgaggac cccgaggtga agttcaactg
     2701 gtacgtggat ggcgtggagg tgcacaatgc caagaccaag ccaagggagg agcagtacaa
     2761 ctctacctat cgcgtggtga gcgtgctgac agtgctgcac caggactggc tgaacggcaa
     2821 ggagtacaag tgtaaggtgt ctaataaggc cctgcctgcc ccaatcgaga agaccatcag
     2881 caaggcaaag ggacagccaa gggagcctca ggtgtatacc ctgccaccca gccgcgacga
     2941 gctgacaaag aaccaggtgt ccctgtactg cctggtgaag ggcttctatc cttccgatat
     3001 cgccgtggag tgggagtcta atggccagcc agagaacaat tacaagacca cacctccagt
     3061 gctggactcc gatggctctt tctttctgta ttccaagctg accgtggata agagccggtg
     3121 gcagcagggc aacgtgttta gctgttccgt gatgcacgag gccctgcaca atcactacac
     3181 acagaagtct ctgagcctgt cccccggcaa gggaggagga ggcagcggcg gaggaggctc
     3241 cggaggcggc ggctctggcg gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt
     3301 gttcctgttt ccacctaagc ccaaagatac cctgatgatc tccaggaccc ctgaagtcac
     3361 ctgcgtggtc gtggacgtgt ctcatgagga ccccgaagtc aagttcaact ggtacgtgga
     3421 cggagtcgaa gtccataacg ccaagaccaa gcctagagaa gagcagtaca actccaccta
     3481 tagagtggtg tctgtgctga cagtgctgca tcaggattgg ctgaatggca aggaatataa
     3541 gtgtaaggtg agcaataagg ccctgcccgc ccctatcgag aagaccatct ctaaagccaa
     3601 aggacagcca agggagccac aggtgtatac actgccaccc agccgggacg agctgaccaa
     3661 gaaccaggtg agcctgacat gtctggtcaa gggcttctac cccagtgaca ttgctgtgga
     3721 gtgggagtcc aatggccagc ctgagaacaa ttataaaact accccaccag tgctggacag
     3781 cgatggctcc ttctttctga ccagcaagct gacagtggat aagtcccgct ggcagcaggg
     3841 aaacgtgttt tcttgtagcg tgatgcatga ggccctgcat aaccactaca cccagaagtc
     3901 cctgtctctg agcccaggca agggaggcag cggaggcgga ggcagcggag gcctggaggt
     3961 gctgtttcag ggccctcagt gcgtgaacct gaccacccga acccagctcc caccagccta
     4021 caccaacagc tttacacggg gcgtgtacta ccctgacaag gtgttcagat ctagcgtcct
     4081 gcacagcact caggacctct tcctgccgtt cttcagcaac gtgacatggt tccacgccat
     4141 ccacgtgagc ggcacaaacg gaaccaagcg gtttgataac cccgtcctgc cattcaatga
     4201 tggagtttac ttcgccagta tcgagaagag taacatcatc cggggctgga tcttcggcac
     4261 caccctggat agcaaaacac agagcctcct gatcgtgaac aatgccacga acgtcgtgat
     4321 caaggtgtgc gagttccagt tttgcaatga tcctttcctg gacgtgtact accacaagaa
     4381 caacaagagc tggatgaaga gcgagttcag agtctacagc agcgccaaca actgcacatt
     4441 cgagtacgtc tctcagcctt ttctgatgga ccttgagggg aaacaaggca acttcaagaa
     4501 cctgagagaa ttcgtgttca agaacatcga cggctacttc aaaatctact ccaagcacac
     4561 acccatcaac ctggtccggg acctccctca gggcttcagc gccctggaac ccctggtcga
     4621 cctgcccata ggcatcaaca taacgcggtt ccaaaccctg ctggccctgc atagatccta
     4681 cctgactcct ggcgacagca gcagcggatg gaccgccgga gctgcagcct actatgtggg
     4741 ctacctgcaa cctagaacct tcctgctgaa gtacaacgag aacggcacaa tcacagacgc
     4801 cgtcgactgc gccctggacc ctctctctga gacaaagtgc accctgaagt ccttcaccgt
     4861 ggaaaagggc atctaccaga ccagcaactt ccgggtgcag cctacagaga gcatcgtgcg
     4921 atttccaaac attaccaacc tctgcccctt cggcgaggtg tttaacgcca caagatttgc
     4981 ctccgtttac gcctggaata gaaagagaat cagcaattgt gtggccgact actccgtgct
     5041 gtataacagc gcctctttca gcaccttcaa gtgctacggc gtttccccaa caaagctgaa
     5101 tgacctgtgc ttcaccaacg tgtacgccga ctccttcgta attagaggcg atgaggtgcg
     5161 gcagatcgca ccaggccaga ccggtaagat cgctgactac aactataagc tgcctgatga
     5221 ttttacaggc tgcgtgatcg cctggaactc taacaacctg gatagcaagg tgggcggcaa
     5281 ctacaactac agataccggc tgtttcgcaa gtctaacctg aaacctttcg agagagacat
     5341 ctccacagag atctaccagg ccggttctac accttgtaac ggggtgcagg gcttcaactg
     5401 ttacttccct ctgcaaagct acggcttcca gcctaccaat ggagtcggct accagccata
     5461 ccgggtggtc gtgctgtcct tcgagttact ccacgccccc gccaccgtct gcggtcctaa
     5521 gaagtccacc aatctggtta agaacaaatg cgtgaacttc aacttcaacg gcctgaccgg
     5581 gaccggcgtg ctgaccgaaa gcaacaaaaa gttcctcccc ttccagcagt tcggccgtga
     5641 tatcgctgac accacagatg ccgtcagaga tccacagacc ctggaaatcc tggatattac
     5701 accctgctcc ttcggaggag tttctgtgat cacccccggg accaatacca gcaaccaggt
     5761 ggctgtgctg taccaaggtg ttaactgcac cgaggttcct gtggccatcc acgccgatca
     5821 gctgacacct acttggagag tgtactccac tggctccaat gtgttccaga ccagggccgg
     5881 atgtctgatc ggcgccgagc acgtgaataa cagttacgag tgcgacatcc ctatcggcgc
     5941 cggcatctgt gccagctacc agacccagac aaacagcaga gggtctgctt cctctgtagc
     6001 tagccagagc atcatcgcct acaccatgag cctgggcgca gagaacagcg tggcctattc
     6061 caacaactct atcgccattc ccaccaactt tacaattagc gtcacaacag agatcctgcc
     6121 cgtgagcatg accaagacca gcgtggactg tacaatgtac atctgtggcg acagcactga
     6181 atgcagcaac ctgctgctgc aatacggctc cttttgcacc caactgaacc gggcgctgac
     6241 cggaatcgcc gtggaacagg acaaaaatac ccaggaggtg ttcgcccaag tgaagcagat
     6301 ctacaagacc ccacctatca aggacttcgg cggctttaac tttagccaga ttctccctga
     6361 tccttctaag cctagcaagc ggagctttat cgaggatctg ctgttcaaca aggtcaccct
     6421 ggccgatgcc ggctttatca aacagtatgg cgattgcctg ggcgacatag ccgccagaga
     6481 tctgatctgc gcccagaaat tcaacggcct gacagttctc ccacctctgc tgaccgacga
     6541 gatgatcgct cagtacacct ctgccctgct ggctggcacc atcacatctg ggtggacatt
     6601 tggcgccggc gccgccctgc agatcccctt tgccatgcag atggcctata gattcaacgg
     6661 aatcggcgtg acccagaacg tgctgtatga aaaccagaag ctgatcgcta accagttcaa
     6721 ttctgccatc ggcaagatcc aggactccct ctcctctacc gccagcgccc tgggcaaact
     6781 gcaggacgtg gtgaatcaga acgcccaagc cctgaacacc ctggtgaagc agctcagcag
     6841 caattttggc gccatcagct ctgtgctgaa cgatatcctg tctagactgg accctccaga
     6901 agccgaagtc cagatcgata gactgatcac aggcagactg cagtccctgc aaacctacgt
     6961 gacccaacag ctgatcaggg ccgctgaaat aagagccagc gccaatctcg ccgctaccaa
     7021 gatgtccgag tgtgtgctgg gacagtctaa acgcgttgac ttctgcggca aaggctatca
     7081 cctgatgagc ttcccccaga gcgcgccgca cggcgtggtg ttcctgcatg tgacatacgt
     7141 gcctgcccac gagaagaatt tcacaaccgc ccctgccatc tgccacgacg gcaaggccca
     7201 cttcccaaga gagggcgttt tcgtttccaa tggcacacac tggttcgtga cacaaagaaa
     7261 cttctacgaa ccccagatta tcaccaccga caacaccttc gtgagtggca attgtgacgt
     7321 ggtcatcgga atcgtgaaca acacagtgta cgaccctctg caacctgagc tggactcttt
     7381 taaggaagag ctggacaagt actttaaaaa ccacaccagc cccgatgtgg acctgggcga
     7441 catcagtggc attaacgcca gcgtggtgaa catccaaaag gaaatcgaca gactgaacga
     7501 ggtggccaag aacctgaacg agtccctgat cgacctgcag gagctcggca aatacgagca
     7561 gggatctgga tacatccccg aggcccccag agatggccag gcctacgtgc ggaaggacgg
     7621 cgagtgggta ctgctgagca cattcctggg cagatccgga ggaggcctgg tgccacagca
     7681 gagcggaggc ctgaacgaca tcttcgaggc ccagaagatc gagtggcacg agggctgatg
     7741 aggatccaga tctgctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     7801 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7861 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7921 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg
     7981 tacccaggtg ctgaagaatt gacccggttc ctcctgggcc agaaagaagc aggcacatcc
     8041 ccttctctgt gacacaccct gtccacgccc ctggttctta gttccagccc cactcatagg
     8101 acactcatag ctcaggaggg ctccgccttc aatcccaccc gctaaagtac ttggagcggt
     8161 ctctccctcc ctcatcagcc caccaaacca aacctagcct ccaagagtgg gaagaaatta
     8221 aagcaagata ggctattaag tgcagaggga gagaaaatgc ctccaacatg tgaggaagta
     8281 atgagagaaa tcatagaatt ttaaggccat gatttaaggc catcatggcc ttaatcttcc
     8341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     8401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     8461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     8521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     8581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactcgg gggggggggg
     9361 cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca
     9421 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     9481 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     9541 atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag
     9601 tcagcgtaat gctctgccag tgttacaacc aattaaccaa ttctgatt
//
