LOCUS       sequence-297294-        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6299)
  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"
COMMENT     Sequence Label: pGBW-m4252787
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             119..139
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             143..208
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    261..382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(272..291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     326..345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      497..543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      573..709
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    754..825
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1051..1639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1131..1150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2139..3348
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3185..3204
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3352..3951
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3352..3371)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3848..3868
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3978..4026
                     /note="synthetic polyadenylation signal"
     CDS             join(4292..4360,4361..5152)
                     /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     complement(4510..4529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5165..5301
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5453..5524
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5529..5575)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5593..5641
                     /note="synthetic polyadenylation signal"
     enhancer        5722..6025
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6027..6229
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6180..6200
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6298..6299,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggc tatataaacg ttttcgcttt tccgtttacg atatatagtc tactcttgtg
       61 cagaatgaat tctcgtaact acatagcaca agtagatgta gttaacttta atctcacaga
      121 aaacctttac tttcagtcga tggattataa ggaccacgat ggagactaca aagatcatga
      181 cattgattac aaggacgacg atgacaaata agctagcttg actgactgag atacagcgta
      241 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      421 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
      481 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      541 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
      601 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
      661 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
      721 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
      781 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
      841 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
      901 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
      961 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1861 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     1921 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     1981 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2041 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2101 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     2161 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     2221 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     2281 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     2341 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     2401 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     2461 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     2521 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     2581 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     2641 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     2701 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     2761 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     2821 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     2881 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     2941 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     3001 tataatgcag ggtggggcca ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg
     3061 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     3121 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     3181 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     3241 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     3301 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     3361 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3421 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3481 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3541 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3601 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3661 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3721 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3781 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3841 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3901 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     3961 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     4021 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     4081 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     4141 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     4201 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     4261 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     4321 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4381 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     4441 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     4501 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4561 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     4621 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4681 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     4741 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4801 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     4861 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     4921 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4981 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5041 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5101 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     5161 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     5221 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     5281 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     5341 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     5401 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     5461 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     5521 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     5581 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     5641 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     5701 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5761 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5821 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5881 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     5941 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6001 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     6061 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6121 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6181 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6241 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
