LOCUS       sequence-297250-        7065 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 7065)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7065)
  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-m4252949
FEATURES             Location/Qualifiers
     source          1..7065
                     /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             488..505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    598..719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(609..628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     663..682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      834..880
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      910..1046
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1091..1162
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1388..1976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1468..1487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2476..3685
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3522..3541
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3689..4717
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4744..4792
                     /note="synthetic polyadenylation signal"
     CDS             join(5058..5126,5127..5918)
                     /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(5276..5295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5931..6067
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6219..6290
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6295..6341)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6359..6407
                     /note="synthetic polyadenylation signal"
     enhancer        6488..6791
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6793..6995
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6946..6966
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7064..7065,1..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 tgagtgacaa aatcattcat ctaaccgatg actcctttga
      181 tacagacgtg ttgaaggccg atggtgcgat actggtagat ttctgggctg aatggtgtgg
      241 cccgtgcaag atgatcgcac ccattctgga cgagatcgcc gacgagtatc aaggaaaatt
      301 aactgtcgct aagcttaata tagatcagaa cccagggacg gcacctaaat acggcatcag
      361 gggaatacct accctcctgc tctttaagaa tggggaagtt gccgcgacaa aagtgggagc
      421 cctgtctaag ggccagctaa aagagttctt ggacgctaac ctggcaggtt caggctcggg
      481 gcacatgcac catcaccatc accatagcag cgggctcgtg ccacgcggct aaggatccgc
      541 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
      601 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      661 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      721 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      781 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
      841 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
      901 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
      961 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     1021 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
     1081 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
     1141 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
     1201 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     1261 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     1321 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     2221 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     2281 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     2341 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     2401 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2461 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2521 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     2581 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2641 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2701 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2761 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     2821 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     2881 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     2941 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     3001 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     3061 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     3121 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     3181 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     3241 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     3301 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca
     3361 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3421 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3481 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3541 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     3601 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     3661 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     3721 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3781 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3841 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     3901 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     3961 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     4021 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     4081 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     4141 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4201 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4261 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4321 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4381 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4441 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4501 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4561 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4621 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4681 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     4741 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4801 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     4861 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     4921 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     4981 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     5041 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5161 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5221 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5281 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5341 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5461 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5581 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5641 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5881 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     5941 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     6001 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     6061 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     6121 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     6181 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     6241 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     6301 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     6361 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     6421 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6481 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6541 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6601 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6661 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6721 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6781 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6841 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6901 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6961 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     7021 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
