LOCUS       sequence-333799-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SARS-CoV-2 S protein receptor binding domain (RBD) expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  AUTHORS   Sinegubova MV, Orlova NA, Kovnir SV, Dayanova LK, Vorobiev II
  TITLE     High-level expression of the monomeric SARS-CoV-2 S protein RBD 
            320-537 in stably transfected CHO cells by the EEF1A1-based plasmid 
            vector.
  JOURNAL   PLoS One. 2021 Feb 2;16(2):e0242890. doi: 
            10.1371/journal.pone.0242890. eCollection 2021.
  PUBMED    33529230
REFERENCE   2  (bases 1 to 7513)
  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: "2021-02-2"; volume: "16(2):e0242890. doi"; pages: " 
            10.1371/journal.pone.0242890. eCollection 2021"
COMMENT     Sequence Label: pTM-RBDv2
FEATURES             Location/Qualifiers
     source          1..7513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2883..2892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3618..3647
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3660..3689
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    3713..4286
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3736..4288
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3880..3897)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4107..4126
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      4303..4312
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4309..4872
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     primer_bind     complement(5541..5558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5712..6300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5792..5811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6471..7262,7263..7331))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7094..7113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7332..7436)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccccgatcc ccggaacgcc agccgccatc caccgcccgc agcccccgga gcccgcggac
       61 cccgaccccc cgccgctcgc ccgccgttgc gccgctctct gcgggggggc tagggggccg
      121 cggggggaag gccacgcccc ctccactttt tccaggaatg cgcggcatga cccgcccccc
      181 ccatgacccg ccaacacccc cccgatgccc cccgggggtc tttcctgggg ggcctttgtc
      241 agggttgcct gtgtcaccca cgggcccctt ggccacgctt gcctgggtca ccccggagcc
      301 ccgggtcacg tttgccgggg tcagccctgg ggcgcttctt cacgctccat tgccatgcag
      361 aattcccaca tgtggacacc ggacagtagg tcctcaaatg ctccttatta ggttggttca
      421 ataatatcaa ttgtttgtta ctaggcagtg atgttgtaca tctggaggag atctcttgag
      481 cccataatca ggttattagg aataaatact ctaaggctaa aaatgtagct tagtgataag
      541 agtgcttgcc tggtgtgctg agaccctcgg ttccatctcc acaaccccat attccattac
      601 aaaatacctt ttcaccgtcc ctagcattaa gaaacaaaac aacaaagaag tttttctttc
      661 ttctgagatc ctgcccggag aggcatttaa aactggccag ggccaaaaaa aaaaaaaaaa
      721 gaaaaaaaag aaaagaaaac aggctagggc cggcatggtg gcgcacgcct ttaatcccag
      781 cacgcaggag gcagaggcag ggcggatctc tgtgagtttg aggtcagcct ggtctaccta
      841 gtgagtttca gggcacccag ggctaaagag actgtctcaa aaacaaaaca gccacacaat
      901 cagaaccaca gcaaaacgca gttatgatcc ttggaactgt aggaatgaca agcatttaaa
      961 taataggacg agccattttt gagaagctct gatttcacaa gtgtcaggga tgggctctgg
     1021 gcgagtaaga ttgctaatgc tggcctctaa atgagaccac gtggagttga ttagattctt
     1081 ttcatgttcc tcgtgctcta tcaaataact gtacccaaat acacacacac acacacacac
     1141 acaatgcgcg cacacacaaa atcctttttt agcttaagaa gcccagaatc agaagtaaag
     1201 ctaactgtgg gacttaagta ttattctgaa cggaactccc agggcgtgaa gcgcgcttca
     1261 ggcttccaga gaagcagctg gcgctggatg gaatgaacca agaggccagc acaggggcag
     1321 atccgtcgag ctctcggcca ccgagctgag cccttaggtt ctggggctgg gaagggtccc
     1381 taggattgtg cacctctccc gcgggggaca agcaggggga tggcggggct gacgtcggga
     1441 ggtggcctcc acgggaaggg acacccggat ctcgacacag ccttggcagt ggagtcagga
     1501 agggtaggga cagattctgg acgccctctt ggccagctcc tcaccgcccc acccccgact
     1561 ggagccgaga gtaattcata caaaaggagg gatcgccttc gcccctggga atcccaggga
     1621 ccgtcgctaa attctggccg gcctcccagc ccggaaccgc tgtgcccgcc cagcgcggcg
     1681 ggaggagcct gcgcctaggg cggatcgcgg gtcggcggga gagcacaagc ccacagtccc
     1741 cggcggtggg ggaggggcgc gctgagcggg ggcccgggag ccagcgcggg gcaaactggg
     1801 aaagtggcgt cgtgtgctgg ctccgccctc ttcccgaggg tgggggagaa cggtataaaa
     1861 gtgcggtagt cgcgttggac gttctttttc gcaacgggtt tgccgtcaga acgcaggtga
     1921 gtggcgggtg tggcctccgc gggcccgggc tccctccttt gagcggggtc gggccgccgt
     1981 gcgggtgtcg tcggccgggc ttctctgcga gcgttcccgc cctggatggc gggctgtgcg
     2041 ggagggcgag ggggggaggc ctggcggcgg ccccggagcc tcgcctcgtg tcgggcgtga
     2101 ggcctagcgt ggcttccgcc ccgccgcgtg ccaccgcggc cgcgctttgc tgtctgcccg
     2161 gctgccctcg attgcctgcc cgcggcccgg gccaacaaag ggagggcgtg gagctggctg
     2221 gtagggagcc ccgtagtccg catgtcgggc agggagagcg gcagcagtcg ggggggggac
     2281 cgggcccgcc cgtcccgcag cacatgtcca acgccgcctg gacgggtagc ggcctgtgtc
     2341 ctgataaggc ggccgggcgg tgggttttag atgccgggtt caggtggccc cgggtcccgg
     2401 cccggtctgg ccagtacccc gtagtggctt agctccgagg agggcgaggc ccgcccgccc
     2461 ggcaccagtt gcgtgcgcgg aaagatggcc gctcccgggc cctgtagcaa ggagctcaaa
     2521 atggaggacg cggcagcccg gcggagcggg gcgggtgagt cacccacaca aaggaagagg
     2581 gccttgcccc tcgccggccg ctgcttcctg tgaccccgtg gtgtaccggc cgcacttcag
     2641 tcaccccggg cgctctttcg gagcaccgct ggcctccgct gggggagggg atctgtctaa
     2701 tggcgttgga gtttgctcac atttggtggg tggagactgt agccaggcca gcctggccat
     2761 gaaagtaatt cttggaattt gcccattttg agtttggagc gaagctgatt gacaaagctg
     2821 cttagccgtt caaaggtatt cttcgaactt tttttttaag gtgttgtgaa aacctcgagg
     2881 ccgccaccat ggatgcaatg aagagagggc tctgctgtgt gctgctgctg tgtggagcag
     2941 tcttcgtctc ggctagcgtg cagcccaccg aatccatcgt gcggttcccc aatatcacca
     3001 atctgtgccc cttcggcgag gtgttcaatg ccaccagatt cgcctctgtg tacgcctgga
     3061 accggaagcg gatcagcaat tgcgtggccg actactccgt gctgtacaac tccgccagct
     3121 tcagcacctt caagtgctac ggcgtgtccc ctaccaagct gaacgacctg tgcttcacaa
     3181 acgtgtacgc cgacagcttc gtgatccggg gagatgaagt gcggcagatt gcccctggac
     3241 agacaggcaa gatcgccgac tacaactaca agctgcccga cgacttcacc ggctgtgtga
     3301 ttgcctggaa cagcaacaac ctggactcca aagtcggcgg caactacaat tacctgtacc
     3361 ggctgttccg gaagtccaat ctgaagccct tcgagcggga catctccacc gagatctatc
     3421 aggccggcag caccccttgt aacggcgtgg aaggcttcaa ctgctacttc ccactgcagt
     3481 cctacggctt tcagcccaca aatggcgtgg gctatcagcc ctacagagtg gtggtgctga
     3541 gcttcgaact gctgcatgcc cctgccacag tgtgcggccc taagaaaagc accaatctcg
     3601 tgaagaacaa acccggtgaa caaaaactga tcagcgaaga ggatctgtct gcaggcggtc
     3661 atcaccatca ccatcaccat caccatcact agcggccgcg cagttaacgc cgcccctctc
     3721 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     3781 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     3841 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3901 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     3961 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4021 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     4081 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     4141 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     4201 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     4261 ttttcctttg aaaaacacga tgataatatg gccacaagat ctgccaccat ggttcgacca
     4321 ttgaactgca tcgtcgccgt gtcccaaaat atggggattg gcaagaacgg agacctaccc
     4381 tggcctccgc tcaggaacga gttcaagtac ttccaaagaa tgaccacaac ctcttcagtg
     4441 gaaggtaaac agaatctggt gattatgggt aggaaaacct ggttctccat tcctgagaag
     4501 aatcgacctc taaaggacag aattaatata gttctcagta gagaactcaa agaaccacca
     4561 cgaggagctc attttcttgc caaaagtttg gatgatgcct taagacttat tgaacaaccg
     4621 gaattggcaa gtaaagtaga catggtttgg atagtcggag gcagttctgt ttaccaggaa
     4681 gccatgaatc aaccaggcca cctcagactc tttgtgacaa ggatcatgca ggaatttgaa
     4741 agtgacacgt ttttcccaga aattgatttg gggaaatata aacttctccc agaataccca
     4801 ggcgtcctct ctgaggtcca ggaggaaaaa ggcatcaagt ataagtttga agtctacgag
     4861 aagaaagact aatctagcat attaccccta acacctgcca ccccagtctt aatcagtggt
     4921 ggaagaacgg tctcagaact gtttgtctca attggccatt taagtttaat agtgaaagac
     4981 tggttaatga taacaatgca tcggaaaacc ttcaggagga aaggagaatg ttttgtggaa
     5041 catttttgtg tgtgtggcag ttttaagtta ttagttttca aaatcagtac tttttaatgg
     5101 aaacaacttg accaaaaatc tgtcacagaa ttttgagacc cattaaaata caagtttaat
     5161 gagaagtctg tctctgttaa tgctgaagtc attactaagt gcttagctta gcaaggtatg
     5221 tggatgccca tttgtgttcc aagggattgg actgttcatc aggacccaga gctgagtttc
     5281 aagggctcaa gagatggctt attacctgtg ggtgtcttga aggttctggt tgggacaaat
     5341 taggaatgtt tttggcagac atggtgacta cccttcatct gggtgagttc agttgatttg
     5401 tcttgagcct ttggggttta cacaagtaaa tgacatcata cagttagtgt attgttagtg
     5461 aatattaata tatgaggcag gctttgctct agacatatgt cgacgtccgg agctctggcg
     5521 cgccatggct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7441 tttccccgaa aagtgccacc tgcatctcat cacggtcacg catggctgcg cccgcgcagc
     7501 gccccccgac ccc
//
