LOCUS       sequence-338334-        7434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for protein expression studies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7434)
  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 7434)
  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: p1.1-Tr2-eGFP
FEATURES             Location/Qualifiers
     source          1..7434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(60..648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(140..159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(819..1610,1611..1679))
                     /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     1442..1461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1680..1784)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      4744..4753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4750..4752,4753..4755,4756..5469)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4750..4752,4753..4755,4756..5466)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4795..4816)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5056..5075)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5403..5424
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5495..6068
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5518..6070
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5662..5679)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5889..5908
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      6085..6094
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6091..6654
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     primer_bind     complement(7323..7340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
       61 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
      961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1801 gtgccacctg catctcatca cggtcacgca tggctgcgcc cgcgcagcgc cccccgaccc
     1861 ccccccgatc cccggaacgc cagccgccat ccaccgcccg cagcccccgg agcccgcgga
     1921 ccccgacccc ccgccgctcg cccgccgttg cgccgctctc tgcggggggg ctagggggcc
     1981 gcggggggaa ggccacgccc cctccacttt ttccaggaat gcgcggcatg acccgccccc
     2041 cccatgaccc gccaacaccc ccccgatgcc ccccgggggt ctttcctggg gggcctttgt
     2101 cagggttgcc tgtgtcaccc acgggcccct tggccacgct tgcctgggtc accccggagc
     2161 cccgggtcac gtttgccggg gtcagccctg gggcgcttct tcacgctcca ttgccatgca
     2221 gaattcccac atgtggacac cggacagtag gtcctcaaat gctccttatt aggttggttc
     2281 aataatatca attgtttgtt actaggcagt gatgttgtac atctggagga gatctcttga
     2341 gcccataatc aggttattag gaataaatac tctaaggcta aaaatgtagc ttagtgataa
     2401 gagtgcttgc ctggtgtgct gagaccctcg gttccatctc cacaacccca tattccatta
     2461 caaaatacct tttcaccgtc cctagcatta agaaacaaaa caacaaagaa gtttttcttt
     2521 cttctgagat cctgcccgga gaggcattta aaactggcca gggccaaaaa aaaaaaaaaa
     2581 agaaaaaaaa gaaaagaaaa caggctaggg ccggcatggt ggcgcacgcc tttaatccca
     2641 gcacgcagga ggcagaggca gggcggatct ctgtgagttt gaggtcagcc tggtctacct
     2701 agtgagtttc agggcaccca gggctaaaga gactgtctca aaaacaaaac agccacacaa
     2761 tcagaaccac agcaaaacgc agttatgatc cttggaactg taggaatgac aagcatttaa
     2821 ataataggac gagccatttt tgagaagctc tgatttcaca agtgtcaggg atgggctctg
     2881 ggcgagtaag attgctaatg ctggcctcta aatgagacca cgtggagttg attagattct
     2941 tttcatgttc ctcgtgctct atcaaataac tgtacccaaa tacacacaca cacacacaca
     3001 cacaatgcgc gcacacacaa aatccttttt tagcttaaga agcccagaat cagaagtaaa
     3061 gctaactgtg ggacttaagt attattctga acggaactcc cagggcgtga agcgcgcttc
     3121 aggcttccag agaagcagct ggcgctggat ggaatgaacc aagaggccag cacaggggca
     3181 gatccgtcga gctctcggcc accgagctga gcccttaggt tctggggctg ggaagggtcc
     3241 ctaggattgt gcacctctcc cgcgggggac aagcaggggg atggcggggc tgacgtcggg
     3301 aggtggcctc cacgggaagg gacacccgga tctcgacaca gccttggcag tggagtcagg
     3361 aagggtaggg acagattctg gacgccctct tggccagctc ctcaccgccc cacccccgac
     3421 tggagccgag agtaattcat acaaaaggag ggatcgcctt cgcccctggg aatcccaggg
     3481 accgtcgcta aattctggcc ggcctcccag cccggaaccg ctgtgcccgc ccagcgcggc
     3541 gggaggagcc tgcgcctagg gcggatcgcg ggtcggcggg agagcacaag cccacagtcc
     3601 ccggcggtgg gggaggggcg cgctgagcgg gggcccggga gccagcgcgg ggcaaactgg
     3661 gaaagtggcg tcgtgtgctg gctccgccct cttcccgagg gtgggggaga acggtataaa
     3721 agtgcggtag tcgcgttgga cgttcttttt cgcaacgggt ttgccgtcag aacgcaggtg
     3781 agtggcgggt gtggcctccg cgggcccggg ctccctcctt tgagcggggt cgggccgccg
     3841 tgcgggtgtc gtcggccggg cttctctgcg agcgttcccg ccctggatgg cgggctgtgc
     3901 gggagggcga gggggggagg cctggcggcg gccccggagc ctcgcctcgt gtcgggcgtg
     3961 aggcctagcg tggcttccgc cccgccgcgt gccaccgcgg ccgcgctttg ctgtctgccc
     4021 ggctgccctc gattgcctgc ccgcggcccg ggccaacaaa gggagggcgt ggagctggct
     4081 ggtagggagc cccgtagtcc gcatgtcggg cagggagagc ggcagcagtc ggggggggga
     4141 ccgggcccgc ccgtcccgca gcacatgtcc aacgccgcct ggacgggtag cggcctgtgt
     4201 cctgataagg cggccgggcg gtgggtttta gatgccgggt tcaggtggcc ccgggtcccg
     4261 gcccggtctg gccagtaccc cgtagtggct tagctccgag gagggcgagg cccgcccgcc
     4321 cggcaccagt tgcgtgcgcg gaaagatggc cgctcccggg ccctgtagca aggagctcaa
     4381 aatggaggac gcggcagccc ggcggagcgg ggcgggtgag tcacccacac aaaggaagag
     4441 ggccttgccc ctcgccggcc gctgcttcct gtgaccccgt ggtgtaccgg ccgcacttca
     4501 gtcaccccgg gcgctctttc ggagcaccgc tggcctccgc tgggggaggg gatctgtcta
     4561 atggcgttgg agtttgctca catttggtgg gtggagactg tagccaggcc agcctggcca
     4621 tgaaagtaat tcttggaatt tgcccatttt gagtttggag cgaagctgat tgacaaagct
     4681 gcttagccgt tcaaaggtat tcttcgaact ttttttttaa ggtgttgtga aaacctcgag
     4741 gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4801 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4861 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4921 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4981 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5041 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5101 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5161 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     5221 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5281 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5341 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     5401 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5461 tacaagtaag ctagcggccg cgcagttaac gccgcccctc tccctccccc ccccctaacg
     5521 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     5581 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     5641 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     5701 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     5761 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     5821 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     5881 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     5941 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     6001 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     6061 gatgataata tggccacaag atctgccacc atggttcgac cattgaactg catcgtcgcc
     6121 gtgtcccaaa atatggggat tggcaagaac ggagacctac cctggcctcc gctcaggaac
     6181 gagttcaagt acttccaaag aatgaccaca acctcttcag tggaaggtaa acagaatctg
     6241 gtgattatgg gtaggaaaac ctggttctcc attcctgaga agaatcgacc tctaaaggac
     6301 agaattaata tagttctcag tagagaactc aaagaaccac cacgaggagc tcattttctt
     6361 gccaaaagtt tggatgatgc cttaagactt attgaacaac cggaattggc aagtaaagta
     6421 gacatggttt ggatagtcgg aggcagttct gtttaccagg aagccatgaa tcaaccaggc
     6481 cacctcagac tctttgtgac aaggatcatg caggaatttg aaagtgacac gtttttccca
     6541 gaaattgatt tggggaaata taaacttctc ccagaatacc caggcgtcct ctctgaggtc
     6601 caggaggaaa aaggcatcaa gtataagttt gaagtctacg agaagaaaga ctaatctagc
     6661 atattacccc taacacctgc caccccagtc ttaatcagtg gtggaagaac ggtctcagaa
     6721 ctgtttgtct caattggcca tttaagttta atagtgaaag actggttaat gataacaatg
     6781 catcggaaaa ccttcaggag gaaaggagaa tgttttgtgg aacatttttg tgtgtgtggc
     6841 agttttaagt tattagtttt caaaatcagt actttttaat ggaaacaact tgaccaaaaa
     6901 tctgtcacag aattttgaga cccattaaaa tacaagttta atgagaagtc tgtctctgtt
     6961 aatgctgaag tcattactaa gtgcttagct tagcaaggta tgtggatgcc catttgtgtt
     7021 ccaagggatt ggactgttca tcaggaccca gagctgagtt tcaagggctc aagagatggc
     7081 ttattacctg tgggtgtctt gaaggttctg gttgggacaa attaggaatg tttttggcag
     7141 acatggtgac tacccttcat ctgggtgagt tcagttgatt tgtcttgagc ctttggggtt
     7201 tacacaagta aatgacatca tacagttagt gtattgttag tgaatattaa tatatgaggc
     7261 aggctttgct ctagacatat gtcgacgtcc ggagctctgg cgcgccatgg ctcttccgct
     7321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaac
//
