LOCUS       sequence-297424-        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-m4252918
FEATURES             Location/Qualifiers
     source          1..6720
                     /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    253..374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(264..283)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     318..337
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      489..535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      565..701
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    746..817
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1043..1631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1123..1142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2131..3340
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3177..3196
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3344..4372
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4399..4447
                     /note="synthetic polyadenylation signal"
     CDS             join(4713..4781,4782..5573)
                     /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(4931..4950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5586..5722
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5874..5945
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5950..5996)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6014..6062
                     /note="synthetic polyadenylation signal"
     enhancer        6143..6446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6448..6650
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6601..6621
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6719..6720,1..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 cagttaagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
      241 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      301 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      361 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      421 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
      481 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
      541 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
      601 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
      661 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
      721 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
      781 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
      841 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
      901 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
      961 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1021 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1081 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1141 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1201 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1261 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1321 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1381 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1441 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1501 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1561 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1621 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1681 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1741 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1801 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1861 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     1921 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     1981 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     2041 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     2101 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     2161 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     2221 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     2281 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     2341 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     2401 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     2461 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     2521 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     2581 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     2641 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     2701 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     2761 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     2821 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     2881 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     2941 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3001 agggtggggc caccagccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3061 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     3121 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     3181 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     3241 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     3301 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     3361 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     3421 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     3481 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     3541 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     3601 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     3661 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     3721 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     3781 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     3841 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     3901 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     3961 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4021 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4081 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     4141 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     4201 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     4261 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     4321 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     4381 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     4441 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     4501 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     4561 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     4621 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     4681 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     4741 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4801 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     4861 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     4921 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4981 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     5041 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5101 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     5161 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5221 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     5281 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     5341 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5401 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5461 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5521 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     5581 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     5641 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     5701 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     5761 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     5821 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     5881 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     5941 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     6001 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     6061 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     6121 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6181 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6241 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6301 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6361 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6421 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     6481 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6541 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6601 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     6661 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
