LOCUS       sequence-313666-        6257 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 6257)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6257)
  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-m4252830
FEATURES             Location/Qualifiers
     source          1..6257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             697..717
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             718..744
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    797..918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(808..827)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     862..881
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1033..1079
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1109..1245
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1290..1361
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1587..2175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1667..1686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2675..3884
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3721..3740
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3888..4487
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3888..3907)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4384..4404
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4514..4562
                     /note="synthetic polyadenylation signal"
     CDS             join(4828..4896,4897..5688)
                     /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(5046..5065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5701..5837
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5989..6060
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6065..6111)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6129..6177
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgggcta tataaacgtt
      601 ttcgcttttc cgtttacgat atatagtcta ctcttgtgca gaatgaattc tcgtaactac
      661 atagcacaag tagatgtagt taactttaat ctcacagaaa acctttactt tcagtcgtac
      721 ccctatgacg taccagatta tgcgtaagct agcttgactg actgagatac agcgtacctt
      781 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
      841 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      901 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
      961 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1021 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1081 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1141 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     1201 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     1261 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     1321 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     1381 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     1441 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     1501 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     1561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2401 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     2461 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     2521 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     2581 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     2641 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     2701 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     2761 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     2821 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     2881 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     2941 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3001 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     3061 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     3121 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     3181 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     3241 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     3301 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     3361 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     3421 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     3481 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     3541 atgcagggtg gggccaccag ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     3601 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     3661 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     3721 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     3781 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta
     3841 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca
     3901 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3961 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4021 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4081 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4141 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4201 cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4261 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4321 tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4381 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4441 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag
     4501 cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
     4561 tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     4621 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     4681 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     4741 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     4801 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     4861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4921 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     4981 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     5041 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5101 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     5161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5221 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     5281 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     5401 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     5461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     5701 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     5761 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     5821 cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc
     5881 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     5941 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6001 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6061 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6121 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6181 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6241 ccatatatgg agttccg
//
