LOCUS       sequence-333802-        6862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in CHO cells - hTPA leader and C-terminal c-myc 
            and 10xhis tags are coded.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6862)
  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 6862)
  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
FEATURES             Location/Qualifiers
     source          1..6862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             92..121
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             134..163
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    187..760
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    210..762
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(354..371)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     581..600
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      777..786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             783..1346
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     primer_bind     complement(2015..2032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2186..2774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2266..2285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2945..3736,3737..3805))
                     /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     3568..3587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3806..3910)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgaggccgcc accatggatg caatgaagag agggctctgc tgtgtgctgc tgctgtgtgg
       61 agcagtcttc gtctcggcta gcgctaccgg tgaacaaaaa ctgatcagcg aagaggatct
      121 gtctgcaggc ggtcatcacc atcaccatca ccatcaccat cactagcggc cgcgcagtta
      181 acgccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
      241 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
      301 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
      361 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
      421 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
      481 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
      541 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
      601 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
      661 tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
      721 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca agatctgcca
      781 ccatggttcg accattgaac tgcatcgtcg ccgtgtccca aaatatgggg attggcaaga
      841 acggagacct accctggcct ccgctcagga acgagttcaa gtacttccaa agaatgacca
      901 caacctcttc agtggaaggt aaacagaatc tggtgattat gggtaggaaa acctggttct
      961 ccattcctga gaagaatcga cctctaaagg acagaattaa tatagttctc agtagagaac
     1021 tcaaagaacc accacgagga gctcattttc ttgccaaaag tttggatgat gccttaagac
     1081 ttattgaaca accggaattg gcaagtaaag tagacatggt ttggatagtc ggaggcagtt
     1141 ctgtttacca ggaagccatg aatcaaccag gccacctcag actctttgtg acaaggatca
     1201 tgcaggaatt tgaaagtgac acgtttttcc cagaaattga tttggggaaa tataaacttc
     1261 tcccagaata cccaggcgtc ctctctgagg tccaggagga aaaaggcatc aagtataagt
     1321 ttgaagtcta cgagaagaaa gactaatcta gcatattacc cctaacacct gccaccccag
     1381 tcttaatcag tggtggaaga acggtctcag aactgtttgt ctcaattggc catttaagtt
     1441 taatagtgaa agactggtta atgataacaa tgcatcggaa aaccttcagg aggaaaggag
     1501 aatgttttgt ggaacatttt tgtgtgtgtg gcagttttaa gttattagtt ttcaaaatca
     1561 gtacttttta atggaaacaa cttgaccaaa aatctgtcac agaattttga gacccattaa
     1621 aatacaagtt taatgagaag tctgtctctg ttaatgctga agtcattact aagtgcttag
     1681 cttagcaagg tatgtggatg cccatttgtg ttccaaggga ttggactgtt catcaggacc
     1741 cagagctgag tttcaagggc tcaagagatg gcttattacc tgtgggtgtc ttgaaggttc
     1801 tggttgggac aaattaggaa tgtttttggc agacatggtg actacccttc atctgggtga
     1861 gttcagttga tttgtcttga gcctttgggg tttacacaag taaatgacat catacagtta
     1921 gtgtattgtt agtgaatatt aatatatgag gcaggctttg ctctagacat atgtcgacgt
     1981 ccggagctct ggcgcgccat ggctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2041 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2881 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2941 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3001 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3061 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3121 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3181 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3241 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3301 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3361 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3421 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3481 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3541 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3601 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3661 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3721 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3781 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3841 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3901 gggttccgcg cacatttccc cgaaaagtgc cacctgcatc tcatcacggt cacgcatggc
     3961 tgcgcccgcg cagcgccccc cgaccccccc ccgatccccg gaacgccagc cgccatccac
     4021 cgcccgcagc ccccggagcc cgcggacccc gaccccccgc cgctcgcccg ccgttgcgcc
     4081 gctctctgcg ggggggctag ggggccgcgg ggggaaggcc acgccccctc cactttttcc
     4141 aggaatgcgc ggcatgaccc gcccccccca tgacccgcca acaccccccc gatgcccccc
     4201 gggggtcttt cctggggggc ctttgtcagg gttgcctgtg tcacccacgg gccccttggc
     4261 cacgcttgcc tgggtcaccc cggagccccg ggtcacgttt gccggggtca gccctggggc
     4321 gcttcttcac gctccattgc catgcagaat tcccacatgt ggacaccgga cagtaggtcc
     4381 tcaaatgctc cttattaggt tggttcaata atatcaattg tttgttacta ggcagtgatg
     4441 ttgtacatct ggaggagatc tcttgagccc ataatcaggt tattaggaat aaatactcta
     4501 aggctaaaaa tgtagcttag tgataagagt gcttgcctgg tgtgctgaga ccctcggttc
     4561 catctccaca accccatatt ccattacaaa ataccttttc accgtcccta gcattaagaa
     4621 acaaaacaac aaagaagttt ttctttcttc tgagatcctg cccggagagg catttaaaac
     4681 tggccagggc caaaaaaaaa aaaaaaagaa aaaaaagaaa agaaaacagg ctagggccgg
     4741 catggtggcg cacgccttta atcccagcac gcaggaggca gaggcagggc ggatctctgt
     4801 gagtttgagg tcagcctggt ctacctagtg agtttcaggg cacccagggc taaagagact
     4861 gtctcaaaaa caaaacagcc acacaatcag aaccacagca aaacgcagtt atgatccttg
     4921 gaactgtagg aatgacaagc atttaaataa taggacgagc catttttgag aagctctgat
     4981 ttcacaagtg tcagggatgg gctctgggcg agtaagattg ctaatgctgg cctctaaatg
     5041 agaccacgtg gagttgatta gattcttttc atgttcctcg tgctctatca aataactgta
     5101 cccaaataca cacacacaca cacacacaca atgcgcgcac acacaaaatc cttttttagc
     5161 ttaagaagcc cagaatcaga agtaaagcta actgtgggac ttaagtatta ttctgaacgg
     5221 aactcccagg gcgtgaagcg cgcttcaggc ttccagagaa gcagctggcg ctggatggaa
     5281 tgaaccaaga ggccagcaca ggggcagatc cgtcgagctc tcggccaccg agctgagccc
     5341 ttaggttctg gggctgggaa gggtccctag gattgtgcac ctctcccgcg ggggacaagc
     5401 agggggatgg cggggctgac gtcgggaggt ggcctccacg ggaagggaca cccggatctc
     5461 gacacagcct tggcagtgga gtcaggaagg gtagggacag attctggacg ccctcttggc
     5521 cagctcctca ccgccccacc cccgactgga gccgagagta attcatacaa aaggagggat
     5581 cgccttcgcc cctgggaatc ccagggaccg tcgctaaatt ctggccggcc tcccagcccg
     5641 gaaccgctgt gcccgcccag cgcggcggga ggagcctgcg cctagggcgg atcgcgggtc
     5701 ggcgggagag cacaagccca cagtccccgg cggtggggga ggggcgcgct gagcgggggc
     5761 ccgggagcca gcgcggggca aactgggaaa gtggcgtcgt gtgctggctc cgccctcttc
     5821 ccgagggtgg gggagaacgg tataaaagtg cggtagtcgc gttggacgtt ctttttcgca
     5881 acgggtttgc cgtcagaacg caggtgagtg gcgggtgtgg cctccgcggg cccgggctcc
     5941 ctcctttgag cggggtcggg ccgccgtgcg ggtgtcgtcg gccgggcttc tctgcgagcg
     6001 ttcccgccct ggatggcggg ctgtgcggga gggcgagggg gggaggcctg gcggcggccc
     6061 cggagcctcg cctcgtgtcg ggcgtgaggc ctagcgtggc ttccgccccg ccgcgtgcca
     6121 ccgcggccgc gctttgctgt ctgcccggct gccctcgatt gcctgcccgc ggcccgggcc
     6181 aacaaaggga gggcgtggag ctggctggta gggagccccg tagtccgcat gtcgggcagg
     6241 gagagcggca gcagtcgggg gggggaccgg gcccgcccgt cccgcagcac atgtccaacg
     6301 ccgcctggac gggtagcggc ctgtgtcctg ataaggcggc cgggcggtgg gttttagatg
     6361 ccgggttcag gtggccccgg gtcccggccc ggtctggcca gtaccccgta gtggcttagc
     6421 tccgaggagg gcgaggcccg cccgcccggc accagttgcg tgcgcggaaa gatggccgct
     6481 cccgggccct gtagcaagga gctcaaaatg gaggacgcgg cagcccggcg gagcggggcg
     6541 ggtgagtcac ccacacaaag gaagagggcc ttgcccctcg ccggccgctg cttcctgtga
     6601 ccccgtggtg taccggccgc acttcagtca ccccgggcgc tctttcggag caccgctggc
     6661 ctccgctggg ggaggggatc tgtctaatgg cgttggagtt tgctcacatt tggtgggtgg
     6721 agactgtagc caggccagcc tggccatgaa agtaattctt ggaatttgcc cattttgagt
     6781 ttggagcgaa gctgattgac aaagctgctt agccgttcaa aggtattctt cgaacttttt
     6841 ttttaaggtg ttgtgaaaac ct
//
