LOCUS       sequence-297414-        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-m4252782
FEATURES             Location/Qualifiers
     source          1..6257
                     /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..166
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    219..340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(230..249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     284..303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      455..501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      531..667
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    712..783
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1009..1597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1089..1108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2097..3306
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3143..3162
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3310..3909
                     /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(3310..3329)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3806..3826
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3936..3984
                     /note="synthetic polyadenylation signal"
     CDS             join(4250..4318,4319..5110)
                     /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(4468..4487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5123..5259
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5411..5482
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5487..5533)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5551..5599
                     /note="synthetic polyadenylation signal"
     enhancer        5680..5983
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5985..6187
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6138..6158
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6256..6257,1..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 tttcagtcgt acccctatga cgtaccagat tatgcgtaag ctagcttgac
      181 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      241 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      301 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      361 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      421 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      481 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      541 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      601 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      661 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      721 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      781 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      841 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      901 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
      961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1801 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1861 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1921 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1981 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2041 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2101 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2161 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2221 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2281 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2341 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2401 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2461 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2521 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2581 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2641 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     2701 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     2761 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     2821 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     2881 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     2941 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     3001 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3061 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3121 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3181 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3241 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3301 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3361 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3421 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3481 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3541 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3601 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3661 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3721 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     3781 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3841 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     3901 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     3961 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4021 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4081 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4141 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4201 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4261 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4321 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4381 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4441 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4501 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4561 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4621 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4681 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4741 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4801 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4861 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4921 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4981 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5041 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5101 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5161 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5221 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5281 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5341 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5401 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5461 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5521 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5581 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5641 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5701 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5761 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5821 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5881 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5941 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     6001 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6061 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6121 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6181 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6241 cacccgccag cggccgc
//
