LOCUS       sequence-354484-        9645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.621 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 9645)
  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 9645)
  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.1.621-AVI
FEATURES             Location/Qualifiers
     source          1..9645
                     /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             2828..2851
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6566..6610
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6633..6857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7224..7241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7395..7983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7475..7494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8525..9340)
                     /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(8525..9340)
                     /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     9248..9267
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9332..9351
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9618..9636
                     /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 cgcccaccat
     1381 gggatggtca tgtatcatcc tttttctagt agcaactgca accggcgtac attccgcacc
     1441 agagctgctg ggaggaccta gcgtgttcct gtttccccct aagcctaagg acaccctgat
     1501 gatcagccgg accccagagg tgacatgcgt ggtggtggac gtgtcccacg aggaccccga
     1561 ggtgaagttc aactggtacg tggatggcgt ggaggtgcac aatgccaaga ccaagccaag
     1621 ggaggagcag tacaactcta cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga
     1681 ctggctgaac ggcaaggagt acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat
     1741 cgagaagacc atcagcaagg caaagggaca gccaagggag cctcaggtgt ataccctgcc
     1801 acccagccgc gacgagctga caaagaacca ggtgtccctg tactgcctgg tgaagggctt
     1861 ctatccttcc gatatcgccg tggagtggga gtctaatggc cagccagaga acaattacaa
     1921 gaccacacct ccagtgctgg actccgatgg ctctttcttt ctgtattcca agctgaccgt
     1981 ggataagagc cggtggcagc agggcaacgt gtttagctgt tccgtgatgc acgaggccct
     2041 gcacaatcac tacacacaga agtctctgag cctgtccccc ggcaagggag gaggaggcag
     2101 cggcggagga ggctccggag gcggcggctc tggcggcggc ggcagcgccc ctgaactgct
     2161 gggaggacca agcgtgttcc tgtttccacc taagcccaaa gataccctga tgatctccag
     2221 gacccctgaa gtcacctgcg tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt
     2281 caactggtac gtggacggag tcgaagtcca taacgccaag accaagccta gagaagagca
     2341 gtacaactcc acctatagag tggtgtctgt gctgacagtg ctgcatcagg attggctgaa
     2401 tggcaaggaa tataagtgta aggtgagcaa taaggccctg cccgccccta tcgagaagac
     2461 catctctaaa gccaaaggac agccaaggga gccacaggtg tatacactgc cacccagccg
     2521 ggacgagctg accaagaacc aggtgagcct gacatgtctg gtcaagggct tctaccccag
     2581 tgacattgct gtggagtggg agtccaatgg ccagcctgag aacaattata aaactacccc
     2641 accagtgctg gacagcgatg gctccttctt tctgaccagc aagctgacag tggataagtc
     2701 ccgctggcag cagggaaacg tgttttcttg tagcgtgatg catgaggccc tgcataacca
     2761 ctacacccag aagtccctgt ctctgagccc aggcaaggga ggcagcggag gcggaggcag
     2821 cggaggcctg gaggtgctgt ttcagggccc tcagtgcgtg aacctgacca cccgaaccca
     2881 gctcccacca gcctacacca acagctttac acggggcgtg tactaccctg acaaggtgtt
     2941 cagatctagc gtcctgcaca gcactcagga cctcttcctg ccgttcttca gcaacgtgac
     3001 atggttccac gccatccacg tgagcggcac aaacggaacc aagcggtttg ataaccccgt
     3061 cctgccattc aatgatggag tttacttcgc cagtatcgag aagagtaaca tcatccgggg
     3121 ctggatcttc ggcaccaccc tggatagcaa aacacagagc ctcctgatcg tgaacaatgc
     3181 cacgaacgtc gtgatcaagg tgtgcgagtt ccagttttgc aatgatcctt tcctgggtgt
     3241 gacctccaac cacaagaaca acaagagctg gatggaaagc gagttcagag tctacagcag
     3301 cgccaacaac tgcacattcg agtacgtctc tcagcctttt ctgatggacc ttgaggggaa
     3361 acaaggcaac ttcaagaacc tgagagaatt cgtgttcaag aacatcgacg gctacttcaa
     3421 aatctactcc aagcacacac ccatcaacct ggtccgggac ctccctcagg gcttcagcgc
     3481 cctggaaccc ctggtcgacc tgcccatagg catcaacata acgcggttcc aaaccctgct
     3541 ggccctgcat agatcctacc tgactcctgg cgacagcagc agcggatgga ccgccggagc
     3601 tgcagcctac tatgtgggct acctgcaacc tagaaccttc ctgctgaagt acaacgagaa
     3661 cggcacaatc acagacgccg tcgactgcgc cctggaccct ctctctgaga caaagtgcac
     3721 cctgaagtcc ttcaccgtgg aaaagggcat ctaccagacc agcaacttcc gggtgcagcc
     3781 tacagagagc atcgtgcgat ttccaaacat taccaacctc tgccccttcg gcgaggtgtt
     3841 taacgccaca aagtttgcct ccgtttacgc ctggaataga aagagaatca gcaattgtgt
     3901 ggccgactac tccgtgctgt ataacagcgc ctctttcagc accttcaagt gctacggcgt
     3961 ttccccaaca aagctgaatg acctgtgctt caccaacgtg tacgccgact ccttcgtaat
     4021 tagaggcgat gaggtgcggc agatcgcacc aggccagacc ggtaagatcg ctgactacaa
     4081 ctataagctg cctgatgatt ttacaggctg cgtgatcgcc tggaactcta acaacctgga
     4141 tagcaaggtg ggcggcaact acaactacct gtaccggctg tttcgcaagt ctaacctgaa
     4201 acctttcgag agagacatct ccacagagat ctaccaggcc ggttctacac cttgtaacgg
     4261 ggtgaaaggc ttcaactgtt acttccctct gcaaagctac ggcttccagc ctacctacgg
     4321 agtcggctac cagccatacc gggtggtcgt gctgtccttc gagttactcc acgcccccgc
     4381 caccgtctgc ggtcctaaga agtccaccaa tctggttaag aacaaatgcg tgaacttcaa
     4441 cttcaacggc ctgaccggga ccggcgtgct gaccgaaagc aacaaaaagt tcctcccctt
     4501 ccagcagttc ggccgtgata tcgctgacac cacagatgcc gtcagagatc cacagaccct
     4561 ggaaatcctg gatattacac cctgctcctt cggaggagtt tctgtgatca cccccgggac
     4621 caataccagc aaccaggtgg ctgtgctgta ccaaggtgtt aactgcaccg aggttcctgt
     4681 ggccatccac gccgatcagc tgacacctac ttggagagtg tactccactg gctccaatgt
     4741 gttccagacc agggccggat gtctgatcgg cgccgagcac gtgaataaca gttacgagtg
     4801 cgacatccct atcggcgccg gcatctgtgc cagctaccag acccagacaa acagccacgg
     4861 gtctgcttcc tctgtagcta gccagagcat catcgcctac accatgagcc tgggcgcaga
     4921 gaacagcgtg gcctattcca acaactctat cgccattccc accaacttta caattagcgt
     4981 cacaacagag atcctgcccg tgagcatgac caagaccagc gtggactgta caatgtacat
     5041 ctgtggcgac agcactgaat gcagcaacct gctgctgcaa tacggctcct tttgcaccca
     5101 actgaaccgg gcgctgaccg gaatcgccgt ggaacaggac aaaaataccc aggaggtgtt
     5161 cgcccaagtg aagcagatct acaagacccc acctatcaag gacttcggcg gctttaactt
     5221 tagccagatt ctccctgatc cttctaagcc tagcaagcgg agctttatcg aggatctgct
     5281 gttcaacaag gtcaccctgg ccgatgccgg ctttatcaaa cagtatggcg attgcctggg
     5341 cgacatagcc gccagagatc tgatctgcgc ccagaaattc aacggcctga cagttctccc
     5401 acctctgctg accgacgaga tgatcgctca gtacacctct gccctgctgg ctggcaccat
     5461 cacatctggg tggacatttg gcgccggcgc cgccctgcag atcccctttg ccatgcagat
     5521 ggcctataga ttcaacggaa tcggcgtgac ccagaacgtg ctgtatgaaa accagaagct
     5581 gatcgctaac cagttcaatt ctgccatcgg caagatccag gactccctct cctctaccgc
     5641 cagcgccctg ggcaaactgc agaacgtggt gaatcagaac gcccaagccc tgaacaccct
     5701 ggtgaagcag ctcagcagca attttggcgc catcagctct gtgctgaacg atatcctgtc
     5761 tagactggac cctccagaag ccgaagtcca gatcgataga ctgatcacag gcagactgca
     5821 gtccctgcaa acctacgtga cccaacagct gatcagggcc gctgaaataa gagccagcgc
     5881 caatctcgcc gctaccaaga tgtccgagtg tgtgctggga cagtctaaac gcgttgactt
     5941 ctgcggcaaa ggctatcacc tgatgagctt cccccagagc gcgccgcacg gcgtggtgtt
     6001 cctgcatgtg acatacgtgc ctgcccaaga gaagaatttc acaaccgccc ctgccatctg
     6061 ccacgacggc aaggcccact tcccaagaga gggcgttttc gtttccaatg gcacacactg
     6121 gttcgtgaca caaagaaact tctacgaacc ccagattatc accaccgaca acaccttcgt
     6181 gagtggcaat tgtgacgtgg tcatcggaat cgtgaacaac acagtgtacg accctctgca
     6241 acctgagctg gactctttta aggaagagct ggacaagtac tttaaaaacc acaccagccc
     6301 cgatgtggac ctgggcgaca tcagtggcat taacgccagc gtggtgaaca tccaaaagga
     6361 aatcgacaga ctgaacgagg tggccaagaa cctgaacgag tccctgatcg acctgcagga
     6421 gctcggcaaa tacgagcagg gatctggata catccccgag gcccccagag atggccaggc
     6481 ctacgtgcgg aaggacggcg agtgggtact gctgagcaca ttcctgggca gatccggagg
     6541 aggcctggtg ccacagcaga gcggaggcct gaacgacatc ttcgaggccc agaagatcga
     6601 gtggcacgag ggctgatgag gatccagatc tgctgtgcct tctagttgcc agccatctgt
     6661 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6721 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6781 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     6841 tgcggtgggc tctatgggta cccaggtgct gaagaattga cccggttcct cctgggccag
     6901 aaagaagcag gcacatcccc ttctctgtga cacaccctgt ccacgcccct ggttcttagt
     6961 tccagcccca ctcataggac actcatagct caggagggct ccgccttcaa tcccacccgc
     7021 taaagtactt ggagcggtct ctccctccct catcagccca ccaaaccaaa cctagcctcc
     7081 aagagtggga agaaattaaa gcaagatagg ctattaagtg cagagggaga gaaaatgcct
     7141 ccaacatgtg aggaagtaat gagagaaatc atagaatttt aaggccatga tttaaggcca
     7201 tcatggcctt aatcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7501 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7801 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8041 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8221 tgactcgggg ggggggggcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     8281 caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct
     8341 ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg
     8401 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     8461 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     8521 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     8581 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     8641 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     8701 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     8761 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     8821 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     8881 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     8941 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     9001 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     9061 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     9121 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     9181 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     9241 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     9301 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     9361 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     9421 atcagagatt ttgagacaca acgtggcttt cccccccccc ccattattga agcatttatc
     9481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9541 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     9601 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
