LOCUS       sequence-297576-        6654 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 6654)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6654)
  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-m4252990
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    187..308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(198..217)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     252..271
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      423..469
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      499..635
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    680..751
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(977..1565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1057..1076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2065..3274
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3111..3130
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3278..4306
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4333..4381
                     /note="synthetic polyadenylation signal"
     CDS             join(4647..4715,4716..5507)
                     /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(4865..4884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5520..5656
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5808..5879
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5884..5930)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5948..5996
                     /note="synthetic polyadenylation signal"
     enhancer        6077..6380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6382..6584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6535..6555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6653..6654,1..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 acctaactta
      121 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
      181 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      241 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      301 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
      361 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
      421 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
      481 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
      541 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
      601 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
      661 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
      721 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
      781 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
      841 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
      901 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      961 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1021 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1081 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1141 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1201 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1261 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1321 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1381 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1441 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1501 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1561 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1621 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1681 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1741 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1801 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     1861 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     1921 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     1981 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     2041 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     2101 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     2161 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     2221 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     2281 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     2341 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     2401 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     2461 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     2521 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     2581 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     2641 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     2701 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     2761 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     2821 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     2881 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     2941 gggccaccag ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     3001 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     3061 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     3121 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     3181 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     3241 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     3301 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3361 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     3421 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     3481 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     3541 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3601 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     3661 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     3721 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     3781 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     3841 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     3901 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     3961 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     4021 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     4081 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     4141 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4201 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     4261 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     4321 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     4381 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     4441 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     4501 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     4561 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     4621 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     4681 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4741 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     4801 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     4861 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4921 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     4981 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5041 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     5101 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5161 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     5221 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     5281 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5341 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5401 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5461 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     5521 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     5581 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     5641 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     5701 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     5761 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     5821 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     5881 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     5941 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     6001 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     6061 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6361 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     6421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     6481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     6541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     6601 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
