LOCUS       sequence-300726-        6257 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 6257)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6257)
  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-m4252782
FEATURES             Location/Qualifiers
     source          1..6257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    142..213
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(218..264)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    282..330
                     /note="synthetic polyadenylation signal"
     enhancer        411..714
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        716..918
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     869..889
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      987..996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1107..1127
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1128..1154
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1207..1328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1218..1237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1272..1291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1443..1489
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1519..1655
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1700..1771
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1997..2585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2077..2096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3085..4294
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4131..4150
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4298..4897
                     /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(4298..4317)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4794..4814
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4924..4972
                     /note="synthetic polyadenylation signal"
     CDS             join(5238..5306,5307..6098)
                     /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(5456..5475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6111..6247
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
       61 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
      121 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
      181 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
      241 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
      301 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
      361 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      421 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      481 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      541 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      601 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      661 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
      721 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      781 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      841 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      901 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
      961 tccactcgac acacccgcca gcggccgcca ccatggggta cataaatgtt ttcgcctttc
     1021 cattcacgat ctattcactc cttttatgta ggatgaactc tcgcaattac attgctcagg
     1081 tagacgtcgt gaactttaac ctaactgaaa acctttactt tcagtcgtac ccctatgacg
     1141 taccagatta tgcgtaagct agcttgactg actgagatac agcgtacctt cagctcacag
     1201 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1261 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1321 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1381 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     1441 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     1501 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1561 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     1621 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     1681 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     1741 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     1801 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     1861 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     1921 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1981 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2041 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2101 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2161 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2221 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2281 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2341 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2401 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2461 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2521 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2581 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2641 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2701 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2761 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2821 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     2881 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     2941 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     3001 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     3061 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     3121 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     3181 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     3241 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     3301 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     3361 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     3421 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     3481 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     3541 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     3601 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     3661 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     3721 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     3781 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     3841 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     3901 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     3961 gggccaccag ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     4021 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     4081 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     4141 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     4201 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     4261 taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt
     4321 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4381 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4441 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4501 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4561 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4621 gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4681 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4741 ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4801 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     4861 gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta
     4921 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4981 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     5041 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     5101 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     5161 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     5221 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5281 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5341 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5401 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5461 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5521 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5581 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5641 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5701 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5761 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5821 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5881 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5941 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6001 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6061 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     6121 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     6181 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     6241 tccttaagat cccgaat
//
