LOCUS       sequence-300486-        6209 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 6209)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6209)
  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-m4252972
FEATURES             Location/Qualifiers
     source          1..6209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..231
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             379..978
                     /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(379..398)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     875..895
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1005..1053
                     /note="synthetic polyadenylation signal"
     CDS             join(1319..1387,1388..2179)
                     /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(1537..1556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2192..2328
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2480..2551
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2556..2602)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2620..2668
                     /note="synthetic polyadenylation signal"
     enhancer        2749..3052
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3054..3256
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3207..3227
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3325..3334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3497..3618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3508..3527)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3562..3581
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3733..3779
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3809..3945
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3990..4061
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4287..4875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4367..4386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5375..6209,1..375)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca gccggtaggt
       61 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
      121 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
      181 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
      241 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
      301 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
      361 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
      421 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
      481 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
      541 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
      601 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
      661 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
      721 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
      781 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
      841 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
      901 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
      961 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     1021 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     1081 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     1141 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     1201 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     1261 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     1321 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1381 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     1441 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1501 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1561 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1621 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1681 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     1741 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1801 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1861 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     1921 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1981 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2041 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2101 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2161 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     2221 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     2281 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     2341 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     2401 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     2461 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     2521 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     2581 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     2641 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     2701 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2761 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2821 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2881 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2941 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3001 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     3061 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     3121 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     3181 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     3241 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
     3301 cactcgacac acccgccagc ggccgccacc atggggtaca taaatgtttt cgcctttcca
     3361 ttcacgatct attcactcct tttatgtagg atgaactctc gcaattacat tgctcaggta
     3421 gacgtcgtga actttaacct aacttaagct agcttgactg actgagatac agcgtacctt
     3481 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3541 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3601 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3661 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     3721 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     3781 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     3841 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     3901 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     3961 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     4021 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     4081 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     4141 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     4201 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     4261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5041 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5101 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     5161 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     5221 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     5281 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     5341 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     5401 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     5461 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     5521 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     5581 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     5641 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     5701 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     5761 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     5821 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     5881 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     5941 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     6001 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     6061 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     6121 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     6181 gtacctttgg gagcgcgcgc cctcgtcgt
//
