LOCUS       sequence-297248-        6275 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 6275)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6275)
  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-m4252815
FEATURES             Location/Qualifiers
     source          1..6275
                     /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             140..184
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    237..358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(248..267)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     302..321
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      473..519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      549..685
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    730..801
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1027..1615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1107..1126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2115..3324
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3161..3180
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3328..3927
                     /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(3328..3347)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3824..3844
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3954..4002
                     /note="synthetic polyadenylation signal"
     CDS             join(4268..4336,4337..5128)
                     /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(4486..4505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5141..5277
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5429..5500
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5505..5551)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5569..5617
                     /note="synthetic polyadenylation signal"
     enhancer        5698..6001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6003..6205
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6156..6176
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6274..6275,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggg tacataaatg ttttcgcctt tccattcacg atctattcac tccttttatg
       61 taggatgaac tctcgcaatt acattgctca ggtagacgtc gtgaacttta acctaactga
      121 aaacctttac tttcagtcga aagaaaccgc ggcagctaag ttcgagaggc agcacatgga
      181 cagttaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      241 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      301 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      361 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      421 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
      541 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
      601 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
      661 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
      721 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
      781 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
      841 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
      901 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
      961 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1441 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     1861 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     1921 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     1981 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2041 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2101 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     2161 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     2221 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     2281 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     2341 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     2401 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     2461 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     2521 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     2581 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     2641 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     2701 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     2761 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     2821 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     2881 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     2941 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccaccag
     3001 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     3061 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     3121 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     3181 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     3241 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     3301 ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc
     3361 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3421 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     3481 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     3541 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     3601 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc
     3661 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     3721 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc
     3781 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     3841 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     3901 tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa
     3961 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     4021 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     4081 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     4141 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     4201 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     4261 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4321 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4381 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4441 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4501 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4561 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4621 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4681 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4741 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4801 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     4861 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     4921 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4981 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5041 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5101 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     5161 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     5221 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     5281 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     5341 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     5401 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     5461 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     5521 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     5581 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     5641 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5701 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5761 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     5821 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     5881 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5941 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6001 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     6061 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     6121 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     6181 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     6241 cctaccatcc actcgacaca cccgccagcg gccgc
//
