LOCUS       sequence-300572-        6717 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 6717)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6717)
  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-m4252904
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    10..58
                     /note="synthetic polyadenylation signal"
     terminator      76..122
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    127..198
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(350..486)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(499..1290,1291..1359))
                     /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     1122..1141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1625..1673
                     /note="synthetic polyadenylation signal"
     CDS             complement(1700..2728)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2732..3941)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2876..2895)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4441..5029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4930..4949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5255..5326
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5371..5507)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5537..5583)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5698..5819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5735..5754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5789..5808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5888..5929)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5930..5950)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      6061..6070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6139..6341)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6168..6188)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6343..6646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cagttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
       61 taacgcaacg attgatagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      121 tgaattgaga ggtagctgaa gctgctgtta tgaccactag agggaagaag atacctgtgg
      181 agctaatggt ccaagatgtc gggagccgcg atcgctgcca aaaaggtaag ctgacgatag
      241 ctgacgagat tcgatgcccg acgagcaaat gagcgccacg gctgttcgcg taagctcgaa
      301 acgacggaga ttcgatagag ccagagtcga gtttaaacga ttcgggatct taaggataaa
      361 tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg
      421 aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc
      481 caatgcacct agaatcggtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      721 tttatccgcc tccatccagt ctatcaactg ttgccgggaa gctagagtaa gtagttcgcc
      781 agttaatagt ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca gaagtaagtt
      961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1021 atccgtaaga tgcttttctg tgactggtga atactcaacc aagtcattct gagaatagtg
     1081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1141 cagaacttta aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac tctcaaggat
     1201 cttaccgctg ttgagatcca gttcgatgta acccactcgc gcacccaact gatcttcagc
     1261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1321 aaagggaata agggcgacac ggaaatgttg aatactcatg cgaaagcctc ctgctgatcc
     1381 tgtctcttga tagatgtttg caaaagctag gactcgatta tacgttgacc gatcagacta
     1441 tcaatatcta ggtttaaaca gatcggtacg ctgccgtgac caggcttgag tacaatatct
     1501 gacgattgga cttcctgggt cccaagctgg attccgactg gctgactgcc aagcaacgat
     1561 atggatcgtc gagactcgca ctgagaggat cgttctcggt acaccaggcg cgcctagcgt
     1621 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaag
     1681 actaggctta catactagct tattcctttg ccctcggacg agtgctgggg cgtcggtttc
     1741 cactatcggc gagtacttct acacagccat cggtccagac ggcggcgctt ctgcgggcga
     1801 tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg
     1861 cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa
     1921 tgcggagcat atacgcccgg agccggggcg atcctgcaag ctccggatgc ctccgctcga
     1981 agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga
     2041 cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc
     2101 cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg
     2161 ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg
     2221 tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat
     2281 cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct
     2341 ggctaagatc ggcggcagcg atggcatcca ttgcctccgc gaccggctgc agaacagcgg
     2401 gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat
     2461 aggtcaggct ctcgctgaac tccccaatgt caagcacttc cggaatcggg agcgcggcgg
     2521 atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc
     2581 gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg
     2641 agagctgcat caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg
     2701 tcgcggtgag ttcaggcttt ttacccatgt tgtctaacaa aaaagccaaa aacggccaga
     2761 atttagcgga caatttacaa gtctaacact gaaaattaca tattgaccca aatgattaca
     2821 tttcaaaagg tgcctaaaaa acttcacaaa acacactcgc caaccccgag cgcatagttc
     2881 aaaaccggag cttcagctac ttaagaagat aggtacataa aaccgaccaa agaaactgac
     2941 gcctcactta tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc
     3001 ctaggcccga accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggctggt
     3061 ggccccaccc tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg
     3121 cgctcccaaa ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac
     3181 agacgacaaa ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg
     3241 cccagcccat ctcacccgaa taagagcttt cccgcattag cgaagggctc aagaccttgg
     3301 gttcttgccg cccaccatgc cccccacctt gtttcaacga cctcacagcc cgcctcacaa
     3361 gcgtcttcca ttcaagactc gggaacagcc gccattttgc tgcgctcccc ccaaccccca
     3421 gttcagggca accttgctcg cggacccaga ctacagccct tggcggtctc tccacacgct
     3481 tccgtcccac cgagcggccc ggcggccacg aaagccccgg ccagcccagc agcccgctac
     3541 tcaccaagtg acgatcacag cgatccacaa acaagaaccg cgacccaaat cccggctgcg
     3601 acggaactag ctgtgccaca cccggcgcgt ccttatataa tcatcggcgt tcaccgcccc
     3661 acggagatcc ctccgcagaa tcgccgagaa gggactactt ttcctcgcct gttccgctct
     3721 ctggaaagaa aaccagtgcc ctagagtcac ccaagtcccg tcctaaaatg tccttctgct
     3781 gatactgggg ttctaaggcc gagtcttatg agcagcgggc cgctgtcctg agcgtccggg
     3841 cggaaggatc aggacgctcc tgcgcccttc gtctgacgtg gcagcgctcg ccgtgacgag
     3901 gggggcgccc gcgggaggcg ccaaaacccg gcgcggaggc caggcgcgcc ccgatttcga
     3961 tattcctcgg atctcggata gatcgggatt gaggatgtag gttgaacttc tctcgggtga
     4021 tcccgatctc ggatattcgg gattggagta atacctggcg ataggcgctg ccaagcaata
     4081 gccaaggcgc aaggccggcc aagagacttg cgatagttcg ttcgagaggt agcctgcaag
     4141 atggatcgga cgaaaggttt gaagattttc cgacgagggc cttggagagt tcggaaattt
     4201 aaatgcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4261 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4321 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4381 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4441 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4501 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4561 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4621 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4681 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4741 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4801 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4861 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4921 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4981 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5041 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatga tttaaatgaa
     5101 actcgtaatc ggagaacggg tgcaatagca aggaccagat cgaacttgac tgacgcaatt
     5161 ctgagcgtag tcgaaaccca gacaagtcga tctcgccaag tagctgactt gactggttaa
     5221 ttaagtttcg atagcccaag gtaaccaatg agattaggga gggaagaaag tttgatgaac
     5281 tacttctgac ccctagtggt gtccagaaaa gaccattaaa ggaatgtctt gatcgaaatc
     5341 gagagatcgc ctgcaggata gctgacgacc ttaaggataa atttctggta aggaggacac
     5401 gtatggaagt gggcaagttg gggaagccgt atccgttgct gaatctggca tatgtgggag
     5461 tataagacgc gcagcgtcgc atcaggcatt tttttctgcg ccaatgccgg tagatagcaa
     5521 tccggcccga ggggcacaaa aaacccctca agacccgttt agaggcccca aggggttatg
     5581 ctacgatacc gatagagatg ggccaatacc acatttgtag aggttttact tgctttaaaa
     5641 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     5701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5821 catgtctgtg agctgaaggt acgctgtatc tcagtcagtc aagctagcag aggcctgcgg
     5881 atccttaggt agaatctaag ccgagaagtg gattgggtat aggtttcccc gactgaaagt
     5941 aaaggttttc agttaggtta aagttcacga cgtctacctg agcaatgtaa ttgcgagagt
     6001 tcatcctaca taaaaggagt gaatagatcg tgaatggaaa ggcgaaaaca tttatgtacc
     6061 ccatggtggc ggccgctggc gggtgtgtcg agtggatggt aggatcgaca aagatctgat
     6121 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     6181 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     6241 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     6301 ccgctatcca cgcccattga tgtactgcca aaaccgcatc agcatggtaa tagcgatgac
     6361 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     6421 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     6481 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     6541 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     6601 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     6661 tgggctatga actaatgacc ccgtaattga ttactagatc tggctacact cgatcag
//
