LOCUS       sequence-297371-        4341 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 4341)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4341)
  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-m4133681
FEATURES             Location/Qualifiers
     source          1..4341
                     /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    210..331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(221..240)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     275..294
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      446..492
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      522..658
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    703..774
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1000..1588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1080..1099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2334..2402,2403..3194)
                     /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(2552..2571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3207..3343
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3495..3566
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3571..3617)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3635..3683
                     /note="synthetic polyadenylation signal"
     enhancer        3764..4067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4069..4271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4222..4242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4340..4341,1..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 gctagcttga ctgactgaga
      181 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      241 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      301 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      361 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      421 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
      481 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
      541 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
      601 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
      661 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
      721 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
      781 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
      841 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
      901 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
      961 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1021 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1081 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1141 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1201 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1261 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1321 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1381 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1441 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1501 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1561 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1621 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1681 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1741 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1801 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     1861 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     1921 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     1981 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2041 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2101 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2161 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     2221 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     2281 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     2341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     2461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2521 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2641 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     2761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2881 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     2941 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3181 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     3241 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     3301 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     3361 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     3421 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     3481 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     3541 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     3601 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     3661 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     3721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     3901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     3961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4261 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     4321 gacacacccg ccagcggccg c
//
