LOCUS       sequence-304491-        6693 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 6693)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6693)
  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-m4252894
FEATURES             Location/Qualifiers
     source          1..6693
                     /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..157
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    226..347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(237..256)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     291..310
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      462..508
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      538..674
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    719..790
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1016..1604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1096..1115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2104..3313
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3150..3169
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3317..4345
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4372..4420
                     /note="synthetic polyadenylation signal"
     CDS             join(4686..4754,4755..5546)
                     /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(4904..4923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5559..5695
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5847..5918
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5923..5969)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5987..6035
                     /note="synthetic polyadenylation signal"
     enhancer        6116..6419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6421..6623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6574..6594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6692..6693,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggc tatataaacg ttttcgcttt tccgtttacg atatatagtc tactcttgtg
       61 cagaatgaat tctcgtaact acatagcaca agtagatgta gttaacttta atctcacaga
      121 aaacctttac tttcagtcgc accatcacca tcaccattaa ggatccgcag gcctctgcta
      181 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
      241 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      301 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      361 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      421 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
      481 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
      541 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
      601 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
      661 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
      721 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
      781 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
      841 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
      901 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
      961 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1021 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1081 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1141 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1201 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1261 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1321 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1381 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1441 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1501 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1561 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1621 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1681 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1741 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1801 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     1861 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     1921 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     1981 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     2041 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     2101 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     2161 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     2221 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2281 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     2341 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     2401 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     2461 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     2521 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     2581 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     2641 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     2701 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     2761 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     2821 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     2881 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     2941 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt
     3001 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     3061 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     3121 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     3181 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     3241 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     3301 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3361 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     3421 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     3481 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     3541 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     3601 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     3661 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     3721 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     3781 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     3841 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     3901 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     3961 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4021 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4081 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4141 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4201 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4261 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     4321 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     4381 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     4441 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     4501 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     4561 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     4621 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     4681 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4741 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4801 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4861 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4921 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4981 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5041 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5101 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5161 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5221 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     5281 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     5341 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5401 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5461 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5521 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     5581 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     5641 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     5701 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     5761 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     5821 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     5881 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     5941 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     6001 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     6061 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     6421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     6541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6601 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     6661 taccatccac tcgacacacc cgccagcggc cgc
//
