LOCUS       sequence-299992-        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-m4252977
FEATURES             Location/Qualifiers
     source          1..6209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    171..292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(182..201)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     236..255
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      407..453
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      483..619
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    664..735
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(961..1549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1041..1060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2049..3258
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3095..3114
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3262..3861
                     /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(3262..3281)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3758..3778
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3888..3936
                     /note="synthetic polyadenylation signal"
     CDS             join(4202..4270,4271..5062)
                     /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(4420..4439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5075..5211
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5363..5434
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5439..5485)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5503..5551
                     /note="synthetic polyadenylation signal"
     enhancer        5632..5935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5937..6139
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6090..6110
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6208..6209,1..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 atctcacata
      121 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      181 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      241 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      301 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
      361 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
      421 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
      481 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
      541 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
      601 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
      661 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
      721 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
      781 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
      841 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
      901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     1801 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     1861 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     1921 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     1981 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2041 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     2101 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     2161 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     2221 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     2281 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     2341 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     2401 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     2461 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     2521 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     2581 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     2641 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     2701 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     2761 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     2821 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     2881 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca ccagccggta
     2941 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     3001 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     3061 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     3121 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     3181 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     3241 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3301 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3361 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3421 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3481 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3541 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     3601 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     3661 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     3721 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     3781 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     3841 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     3901 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     3961 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4021 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4081 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4141 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4201 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4261 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4321 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4381 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4441 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4501 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4561 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4621 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4681 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4741 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     4801 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4861 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4921 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4981 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5041 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5101 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5161 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     5221 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     5281 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     5341 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     5401 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     5461 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     5521 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     5581 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5641 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5701 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5761 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     5821 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     5881 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     5941 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6001 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6061 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     6121 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     6181 atccactcga cacacccgcc agcggccgc
//
