LOCUS       sequence-347247-        6837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the B.1.617.2 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 6837)
  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 6837)
  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-NTD-B.1.617.2-AVI
FEATURES             Location/Qualifiers
     source          1..6837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..59
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(97..119)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     219..238
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        389..768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        769..972
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     923..943
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     969..993
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2902..2925
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3826..3870
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3893..4117
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4484..4501)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4655..5243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4735..4754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5785..6600)
                     /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(5785..6600)
                     /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     6508..6527
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6592..6611
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
       61 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      121 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      181 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      241 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      301 cagattggct attggccatt gcatacgttg tatccatatc ataatatgta catttatatt
      361 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa
      421 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      481 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      541 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      601 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      661 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      721 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      781 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      841 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      901 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      961 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     1021 gacctccata gaagacaccg ggaccgatcc agcctccatc ggctcgcatc tctccttcac
     1081 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     1141 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     1201 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     1261 tttgcctgac cctgcttgct caactctagt taacggtgga gggcagtgta gtctgagcag
     1321 tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc
     1381 tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg
     1441 ctctagacca ccatgggatg gtcatgtatc atcctttttc tagtagcaac tgcaaccggt
     1501 gtacattccg caccagagct gctgggagga cctagcgtgt tcctgtttcc ccctaagcct
     1561 aaggacaccc tgatgatcag ccggacccca gaggtgacat gcgtggtggt ggacgtgtcc
     1621 cacgaggacc ccgaggtgaa gttcaactgg tacgtggatg gcgtggaggt gcacaatgcc
     1681 aagaccaagc caagggagga gcagtacaac tctacctatc gcgtggtgag cgtgctgaca
     1741 gtgctgcacc aggactggct gaacggcaag gagtacaagt gtaaggtgtc taataaggcc
     1801 ctgcctgccc caatcgagaa gaccatcagc aaggcaaagg gacagccaag ggagcctcag
     1861 gtgtataccc tgccacccag ccgcgacgag ctgacaaaga accaggtgtc cctgtactgc
     1921 ctggtgaagg gcttctatcc ttccgatatc gccgtggagt gggagtctaa tggccagcca
     1981 gagaacaatt acaagaccac acctccagtg ctggactccg atggctcttt ctttctgtat
     2041 tccaagctga ccgtggataa gagccggtgg cagcagggca acgtgtttag ctgttccgtg
     2101 atgcacgagg ccctgcacaa tcactacaca cagaagtctc tgagcctgtc ccccggcaag
     2161 ggaggaggag gcagcggcgg aggaggctcc ggaggcggcg gctctggcgg cggcggcagc
     2221 gcccctgaac tgctgggagg accaagcgtg ttcctgtttc cacctaagcc caaagatacc
     2281 ctgatgatct ccaggacccc tgaagtcacc tgcgtggtcg tggacgtgtc tcatgaggac
     2341 cccgaagtca agttcaactg gtacgtggac ggagtcgaag tccataacgc caagaccaag
     2401 cctagagaag agcagtacaa ctccacctat agagtggtgt ctgtgctgac agtgctgcat
     2461 caggattggc tgaatggcaa ggaatataag tgtaaggtga gcaataaggc cctgcccgcc
     2521 cctatcgaga agaccatctc taaagccaaa ggacagccaa gggagccaca ggtgtataca
     2581 ctgccaccca gccgggacga gctgaccaag aaccaggtga gcctgacatg tctggtcaag
     2641 ggcttctacc ccagtgacat tgctgtggag tgggagtcca atggccagcc tgagaacaat
     2701 tataaaacta ccccaccagt gctggacagc gatggctcct tctttctgac cagcaagctg
     2761 acagtggata agtcccgctg gcagcaggga aacgtgtttt cttgtagcgt gatgcatgag
     2821 gccctgcata accactacac ccagaagtcc ctgtctctga gcccaggcaa gggaggcagc
     2881 ggaggcggag gcagcggagg cctggaggtg ctgtttcagg gccctcagtg cgtgaacctg
     2941 agaacccgaa cccagctccc accagcctac accaacagct ttacacgggg cgtgtactac
     3001 cctgacaagg tgttcagatc tagcgtcctg cacagcactc aggacctctt cctgccgttc
     3061 ttcagcaacg tgacatggtt ccacgccatc cacgtgagcg gcacaaacgg aaccaagcgg
     3121 tttgataacc ccgtcctgcc attcaatgat ggagtttact tcgccagtac cgagaagagt
     3181 aacatcatcc ggggctggat cttcggcacc accctggata gcaaaacaca gagcctcctg
     3241 atcgtgaaca atgccacgaa cgtcgtgatc aaggtgtgcg agttccagtt ttgcaatgat
     3301 cctttcctgg acgtgtacta ccacaagaac aacaagagct ggatggaaag cggcgtctac
     3361 agcagcgcca acaactgcac attcgagtac gtctctcagc cttttctgat ggaccttgag
     3421 gggaaacaag gcaacttcaa gaacctgaga gaattcgtgt tcaagaacat cgacggctac
     3481 ttcaaaatct actccaagca cacacccatc aacctggtcc gggacctccc tcagggcttc
     3541 agcgccctgg aacccctggt cgacctgccc ataggcatca acataacgcg gttccaaacc
     3601 ctgctggccc tgcatagatc ctacctgact cctggcgaca gcagcagcgg atggaccgcc
     3661 ggagctgcag cctactatgt gggctacctg caacctagaa ccttcctgct gaagtacaac
     3721 gagaacggca caatcacaga cgccgtcgac tgcgccctgg accctctctc tgagacaaag
     3781 tgcaccctga agtccggagg aggcctggtg ccacagcaga gcggaggcct gaacgacatc
     3841 ttcgaggccc agaagatcga gtggcacgag ggctgatgag gatccagatc tgctgtgcct
     3901 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3961 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4021 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4081 aatagcaggc atgctgggga tgcggtgggc tctatgggta cccaggtgct gaagaattga
     4141 cccggttcct cctgggccag aaagaagcag gcacatcccc ttctctgtga cacaccctgt
     4201 ccacgcccct ggttcttagt tccagcccca ctcataggac actcatagct caggagggct
     4261 ccgccttcaa tcccacccgc taaagtactt ggagcggtct ctccctccct catcagccca
     4321 ccaaaccaaa cctagcctcc aagagtggga agaaattaaa gcaagatagg ctattaagtg
     4381 cagagggaga gaaaatgcct ccaacatgtg aggaagtaat gagagaaatc atagaatttt
     4441 aaggccatga tttaaggcca tcatggcctt aatcttccgc ttcctcgctc actgactcgc
     4501 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4561 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4621 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4681 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4741 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4801 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4861 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4921 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4981 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5041 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5101 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5161 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5221 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5281 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5341 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5401 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5461 ttcgttcatc catagttgcc tgactcgggg ggggggggcg ctgaggtctg cctcgtgaag
     5521 aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg
     5581 agccacggtt gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct
     5641 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
     5701 aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     5761 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     5821 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     5881 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     5941 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     6001 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     6061 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     6121 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     6181 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     6241 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     6301 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     6361 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     6421 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     6481 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     6541 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
     6601 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     6661 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt cccccccccc
     6721 ccattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctga
//
