LOCUS       sequence-300746-        6702 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 6702)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6702)
  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-m4252877
FEATURES             Location/Qualifiers
     source          1..6702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    151..222
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(227..273)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    291..339
                     /note="synthetic polyadenylation signal"
     enhancer        420..723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        725..927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     878..898
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      996..1005
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1116..1136
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1137..1163
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1232..1353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1243..1262)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1297..1316
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1468..1514
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1544..1680
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1725..1796
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2022..2610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2102..2121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3110..4319
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4156..4175
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4323..5351
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5378..5426
                     /note="synthetic polyadenylation signal"
     CDS             join(5692..5760,5761..6552)
                     /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(5910..5929)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6565..6701
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
       61 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
      121 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
      181 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
      241 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
      301 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
      361 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      601 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      661 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      721 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      841 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      901 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
      961 atcctaccat ccactcgaca cacccgccag cggccgccac catggggtac ataaatgttt
     1021 tcgcctttcc attcacgatc tattcactcc ttttatgtag gatgaactct cgcaattaca
     1081 ttgctcaggt agacgtcgtg aactttaacc taactgaaaa cctttacttt cagtcgtacc
     1141 cctatgacgt accagattat gcgtaaggat ccgcaggcct ctgctagctt gactgactga
     1201 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     1261 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1321 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1381 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1441 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     1501 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     1561 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     1621 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     1681 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     1741 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     1801 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     1861 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     1921 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     1981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2761 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2821 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     2881 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     2941 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     3001 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     3061 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc
     3121 cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac
     3181 gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct
     3241 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg
     3301 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc
     3361 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg
     3421 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt
     3481 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt
     3541 cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag
     3601 tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc
     3661 ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac
     3721 aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg
     3781 aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg
     3841 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     3901 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     3961 ttctgttggc ttataatgca gggtggggcc accagccggt aggtgtgcgg taggcttttc
     4021 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     4081 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     4141 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     4201 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     4261 agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca
     4321 acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4381 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4441 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4501 aagatcgtta tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg
     4561 acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4621 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa
     4681 tggatgccat cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4741 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4801 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4861 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4921 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     4981 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5041 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5101 cgccccggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5161 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5221 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg
     5281 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5341 caaaggaata agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     5401 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     5461 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     5521 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     5581 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     5641 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     5701 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5761 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     5821 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     5881 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5941 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     6001 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6061 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     6121 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6181 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6241 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6301 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6361 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6421 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6481 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6541 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     6601 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     6661 cacttccata cgtgtcctcc ttaccagaaa tttatcctta ag
//
