LOCUS       sequence-314797-        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"
     rep_origin      439..1027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     928..947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1181..1198
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1565..1789
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(1812..1856)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(5568..5591)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(7500..7524)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(7521..7724)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7550..7570)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7725..8104
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(8255..8274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8374..8396
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8434..8452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8719..8738)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             8730..9545
                     /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             8730..9545
                     /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     complement(8803..8822)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac
       61 caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga
      121 tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag gcagacctca
      181 gcgccccccc cccccgagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      241 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      301 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      361 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      421 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      481 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      541 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      601 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      661 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      721 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      781 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      841 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      901 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      961 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1021 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1081 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1141 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1201 agcggaagat taaggccatg atggccttaa atcatggcct taaaattcta tgatttctct
     1261 cattacttcc tcacatgttg gaggcatttt ctctccctct gcacttaata gcctatcttg
     1321 ctttaatttc ttcccactct tggaggctag gtttggtttg gtgggctgat gagggaggga
     1381 gagaccgctc caagtacttt agcgggtggg attgaaggcg gagccctcct gagctatgag
     1441 tgtcctatga gtggggctgg aactaagaac caggggcgtg gacagggtgt gtcacagaga
     1501 aggggatgtg cctgcttctt tctggcccag gaggaaccgg gtcaattctt cagcacctgg
     1561 gtacccatag agcccaccgc atccccagca tgcctgctat tgtcttccca atcctccccc
     1621 ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag acaatgcgat
     1681 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa
     1741 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagc agatctggat
     1801 cctcatcagc cctcgtgcca ctcgatcttc tgggcctcga agatgtcgtt caggcctccg
     1861 ctctgctgtg gcaccaggcc tcctccggat ctgcccagga atgtgctcag cagtacccac
     1921 tcgccgtcct tccgcacgta ggcctggcca tctctggggg cctcggggat gtatccagat
     1981 ccctgctcgt atttgccgag ctcctgcagg tcgatcaggg actcgttcag gttcttggcc
     2041 acctcgttca gtctgtcgat ttccttttgg atgttcacca cgctggcgtt aatgccactg
     2101 atgtcgccca ggtccacatc ggggctggtg tggtttttaa agtacttgtc cagctcttcc
     2161 ttaaaagagt ccagctcagg ttgcagaggg tcgtacactg tgttgttcac gattccgatg
     2221 accacgtcac aattgccact cacgaaggtg ttgtcggtgg tgataatctg gggttcgtag
     2281 aagtttcttt gtgtcacgaa ccagtgtgtg ccattggaaa cgaaaacgcc ctctcttggg
     2341 aagtgggcct tgccgtcgtg gcagatggca ggggcggttg tgaaattctt ctcttgggca
     2401 ggcacgtatg tcacatgcag gaacaccacg ccgtgcggcg cgctctgggg gaagctcatc
     2461 aggtgatagc ctttgccgca gaagtcaacg cgtttagact gtcccagcac acactcggac
     2521 atcttggtag cggcgagatt ggcgctggct cttatttcag cggccctgat cagctgttgg
     2581 gtcacgtagg tttgcaggga ctgcagtctg cctgtgatca gtctatcgat ctggacttcg
     2641 gcttctggag ggtccagtct agacaggata tcgttcagca cagagctgat ggcgccaaaa
     2701 ttgctgctga gctgcttcac cagggtgttc agggcttggg cgttctgatt caccacgtcc
     2761 tgcagtttgc ccagggcgct ggcggtagag gagagggagt cctggatctt gccgatggca
     2821 gaattgaact ggttagcgat cagcttctgg ttttcataca gcacgttctg ggtcacgccg
     2881 attccgttga atctataggc catctgcatg gcaaagggga tctgcagggc ggcgccggcg
     2941 ccaaatgtcc acccagatgt gatggtgcca gccagcaggg cagaggtgta ctgagcgatc
     3001 atctcgtcgg tcagcagagg tgggagaact gtcaggccgt tgaatttctg ggcgcagatc
     3061 agatctctgg cggctatgtc gcccaggcaa tcgccatact gtttgataaa gccggcatcg
     3121 gccagggtga ccttgttgaa cagcagatcc tcgataaagc tccgcttgct aggcttagaa
     3181 ggatcaggga gaatctggct aaagttaaag ccgccgaagt ccttgatagg tggggtcttg
     3241 tagatctgct tcacttgggc gaacacctcc tgggtatttt tgtcctgttc cacggcgatt
     3301 ccggtcagcg cccggttcag ttgggtgcaa aaggagccgt attgcagcag caggttgctg
     3361 cattcagtgc tgtcgccaca gatgtacatt gtacagtcca cgctggtctt ggtcatgctc
     3421 acgggcagga tctctgttgt gacgctaatt gtaaagttgg tgggaatggc gatagagttg
     3481 ttggaatagg ccacgctgtt ctctgcgccc aggctcatgg tgtaggcgat gatgctctgg
     3541 ctagctacag aggaagcaga cccagggctg tttgtctggg tctggtagct ggcacagatg
     3601 ccggcgccga tagggatgtc gcactcgtaa ctgttattca cgtgctcggc gccgatcaga
     3661 catccggccc tggtctggaa cacattggag ccagtggagt acactctcca agtaggtgtc
     3721 agctgatcgg cgtggatggc cacaggaacc tcggtgcagt taacatcttg gtacagcaca
     3781 gccacctggt tgctggtatt ggtcccgggg gtgatcacag aaactcctcc gaaggagcag
     3841 ggtgtaatat ccaggatttc cagggtctgt ggatctctga cggcatctgt ggtgtcagcg
     3901 atatcacggc cgaactgctg gaaggggagg aactttttgt tgctttcggt cagcacgccg
     3961 gtcccggtca ggccgttgaa gttgaagttc acgcatttgt tcttaaccag attggtggac
     4021 ttcttaggac cgcagacggt ggcgggggcg tggagtaact cgaaggacag cacgaccacc
     4081 cggtatggct ggtagccgac tccattggta ggctggaagc cgtagctttg cagagggaag
     4141 taacagttga agccttccac cccgttacaa ggtgtagaac cggcctggta gatctctgtg
     4201 gagatgtctc tctcgaaagg tttcaggtta gacttgcgaa acagccggta caggtagttg
     4261 tagttgccgc ccaccttgct atccaggttg ttagagttcc aggcgatcac gcagcctgta
     4321 aaatcatcag gcagcttata gttgtagtca gcgatcttac cggtctggcc tggtgcgatc
     4381 tgccgcacct catcgcctct aattacgaag gagtcggcgt acacgttggt gaagcacagg
     4441 tcattcagct ttgttgggga aacgccgtag cacttgaagg tgctgaaaga ggcgctgtta
     4501 tacagcacgg agtagtcggc cacacaattg ctgattctct ttctattcca ggcgtaaacg
     4561 gaggcaaatc ttgtggcgtt aaacacctcg ccgaaggggc agaggttggt aatgtttgga
     4621 aatcgcacga tgctctctgt aggctgcacc cggaagttgc tggtctggta gatgcccttt
     4681 tccacggtga aggacttcag ggtgcacttt gtctcagaga gagggtccag ggcgcagtcg
     4741 acggcgtctg tgattgtgcc gttctcgttg tacttcagca ggaaggttct aggttgcagg
     4801 tagcccacat agtaggctgc agctccggcg gtccatccgc tgctgctgtc gccaggagtc
     4861 aggtaggatc tatgcagggc cagcagggtt tggaaccgcg ttatgttgat gcctatgggc
     4921 aggtcgacca ggggttccag ggcgctgaag ccctgaggga ggtcccggac caggttgatg
     4981 ggtgtgtgct tggagtagat tttgaagtag ccgtcgatgt tcttgaacac gaattctctc
     5041 aggttcttga agttgccttg tttcccctca aggtccatca gaaaaggctg agagacgtac
     5101 tcgaatgtgc agttgttggc gctgctgtag actctgaact cgctttccat ccagctcttg
     5161 ttgttcttgt ggtagtacac acccaggaaa ggatcattgc aaaactggaa ctcgcacacc
     5221 ttgatcacga cgttcgtggc attgttcacg atcaggaggc tctgtgtttt gctatccagg
     5281 gtggtgccga agatccagcc ccggatgatg ttactcttct cggtactggc gaagtaaact
     5341 ccatcattga atggcaggac ggggttatca aaccgcttgg ttccgtttgt gccgctcacg
     5401 tggatggcgt ggaaccatgt cacgttgctg aagaacggca ggaagaggtc ctgagtgctg
     5461 tgcaggacgc tagatctgaa caccttgtca gggtagtaca cgccccgtgt aaagctgttg
     5521 gtgtaggctg gtgggagctg ggttcgggtg gtcaggttca cgcactgagg gccctgaaac
     5581 agcacctcca ggcctccgct gcctccgcct ccgctgcctc ccttgcctgg gctcagagac
     5641 agggacttct gggtgtagtg gttatgcagg gcctcatgca tcacgctaca agaaaacacg
     5701 tttccctgct gccagcggga cttatccact gtcagcttgc tggtcagaaa gaaggagcca
     5761 tcgctgtcca gcactggtgg ggtagtttta taattgttct caggctggcc attggactcc
     5821 cactccacag caatgtcact ggggtagaag cccttgacca gacatgtcag gctcacctgg
     5881 ttcttggtca gctcgtcccg gctgggtggc agtgtataca cctgtggctc ccttggctgt
     5941 cctttggctt tagagatggt cttctcgata ggggcgggca gggccttatt gctcacctta
     6001 cacttatatt ccttgccatt cagccaatcc tgatgcagca ctgtcagcac agacaccact
     6061 ctataggtgg agttgtactg ctcttctcta ggcttggtct tggcgttatg gacttcgact
     6121 ccgtccacgt accagttgaa cttgacttcg gggtcctcat gagacacgtc cacgaccacg
     6181 caggtgactt caggggtcct ggagatcatc agggtatctt tgggcttagg tggaaacagg
     6241 aacacgcttg gtcctcccag cagttcaggg gcgctgccgc cgccgccaga gccgccgcct
     6301 ccggagcctc ctccgccgct gcctcctcct cccttgccgg gggacaggct cagagacttc
     6361 tgtgtgtagt gattgtgcag ggcctcgtgc atcacggaac agctaaacac gttgccctgc
     6421 tgccaccggc tcttatccac ggtcagcttg gaatacagaa agaaagagcc atcggagtcc
     6481 agcactggag gtgtggtctt gtaattgttc tctggctggc cattagactc ccactccacg
     6541 gcgatatcgg aaggatagaa gcccttcacc aggcagtaca gggacacctg gttctttgtc
     6601 agctcgtcgc ggctgggtgg cagggtatac acctgaggct cccttggctg tccctttgcc
     6661 ttgctgatgg tcttctcgat tggggcaggc agggccttat tagacacctt acacttgtac
     6721 tccttgccgt tcagccagtc ctggtgcagc actgtcagca cgctcaccac gcgataggta
     6781 gagttgtact gctcctccct tggcttggtc ttggcattgt gcacctccac gccatccacg
     6841 taccagttga acttcacctc ggggtcctcg tgggacacgt ccaccaccac gcatgtcacc
     6901 tctggggtcc ggctgatcat cagggtgtcc ttaggcttag ggggaaacag gaacacgcta
     6961 ggtcctccca gcagctctgg tgcggaatgt acaccggttg cagttgctac tagaaaaagg
     7021 atgatacatg accatcccat ggtggtctag agcggccgcg atatctgatc acacgtgtcg
     7081 acgacggtga ctgcagaaaa gacccatgga aaggaacagt ctgttagtct gtcagctatt
     7141 atgtctggtg gcgcgcgcgg cagcaacgag tactgctcag actacactgc cctccaccgt
     7201 taactagagt tgagcaagca gggtcaggca aagcgtggag agccggctga gtctaggtag
     7261 gctccaaggg agcgccggac aaaggcccgg tctcgacctg agctttaaac ttacctagac
     7321 ggcggacgca gttcaggagg caccacaggc gggaggcggc agaacgcgac tcaaccggcg
     7381 tggatggcgg cctcaggtag ggcggcgggc gcgtgaagga gagatgcgag ccgatggagg
     7441 ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg
     7501 atctgacggt tcactaaacg agctctgctt atatagacct cccaccgtac acgcctaccg
     7561 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     7621 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     7681 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     7741 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     7801 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     7861 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     7921 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     7981 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     8041 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     8101 tgtcaacatg gcggtaatgt tggacatgag ccaatataaa tgtacatatt atgatatgga
     8161 tacaacgtat gcaatggcca atagccaatc tgatgcggta ttttctcctt acgcatctgt
     8221 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     8281 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     8341 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     8401 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     8461 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     8521 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8581 aataaccctg ataaatgctt caataatggg ggggggggga aagccacgtt gtgtctcaaa
     8641 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     8701 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     8761 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     8821 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     8881 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     8941 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     9001 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     9061 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     9121 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     9181 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     9241 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     9301 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     9361 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     9421 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     9481 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     9541 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     9601 acggcggctt tgttgaataa atcgaacttt tgctgagttg aaggatca
//
