LOCUS       sequence-301665-        6720 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 6720)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6720)
  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-m4252926
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        550..1759
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1596..1615
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1763..2791
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    2818..2866
                     /note="synthetic polyadenylation signal"
     CDS             join(3132..3200,3201..3992)
                     /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(3350..3369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4005..4141
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4293..4364
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4369..4415)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4433..4481
                     /note="synthetic polyadenylation signal"
     enhancer        4562..4865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4867..5069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5020..5040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5138..5147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5258..5278
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5279..5323
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    5392..5513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5403..5422)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5457..5476
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5628..5674
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5704..5840
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5885..5956
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(6182..6720,1..50))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6262..6281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
       61 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
      301 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
      361 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
      421 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
      481 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
      541 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
      601 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
      661 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
      721 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
      781 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
      841 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
      901 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
      961 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     1021 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     1081 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     1141 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     1201 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     1261 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     1321 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     1381 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt
     1441 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     1501 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     1561 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     1621 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     1681 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     1741 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     1801 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     1861 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     1921 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     1981 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     2041 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     2101 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     2161 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     2221 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     2281 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     2341 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     2401 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     2461 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     2521 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     2581 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     2641 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     2701 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     2761 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     2821 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     2881 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2941 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     3001 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     3061 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     3121 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3241 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3301 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3421 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3541 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3661 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3721 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3961 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     4021 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     4081 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     4141 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     4201 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     4261 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     4321 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     4381 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     4441 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     4501 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4561 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4621 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4681 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4741 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4801 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4861 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4921 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4981 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     5041 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     5101 cgatcctacc atccactcga cacacccgcc agcggccgcc accatgggct atataaacgt
     5161 tttcgctttt ccgtttacga tatatagtct actcttgtgc agaatgaatt ctcgtaacta
     5221 catagcacaa gtagatgtag ttaactttaa tctcacagaa aacctttact ttcagtcgaa
     5281 agaaaccgcg gcagctaagt tcgagaggca gcacatggac agttaaggat ccgcaggcct
     5341 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     5401 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5461 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5521 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5581 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     5641 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     5701 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     5761 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     5821 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     5881 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     5941 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     6001 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     6061 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     6121 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
//
