LOCUS       sequence-300751-        6272 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 6272)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6272)
  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-m4252832
FEATURES             Location/Qualifiers
     source          1..6272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    99..147
                     /note="synthetic polyadenylation signal"
     terminator      165..211
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    216..287
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(439..575)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(588..1379,1380..1448))
                     /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     1211..1230
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1714..1762
                     /note="synthetic polyadenylation signal"
     CDS             complement(1789..2388)
                     /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(1872..1892)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2369..2388
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2392..3601)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2536..2555)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4101..4689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4590..4609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4915..4986
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5031..5167)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5197..5243)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5358..5479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5395..5414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5449..5468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5532..5573)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5574..5594)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      5705..5714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5783..5985)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5812..5832)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(5987..6272,1..18)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
       61 gattactaga tctggctaca ctcgatcagc agttagataa taaaatcgct atccatcgaa
      121 gatggatgtg tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg
      181 ggcctctaaa cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat
      241 gaccactaga gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga
      301 tcgctgccaa aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg
      361 agcgccacgg ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag
      421 tttaaacgat tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
      481 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
      541 agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta
      601 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      661 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      721 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      781 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt
      841 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt
      901 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      961 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1021 ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1081 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa
     1141 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1201 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag
     1261 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1321 cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1381 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1441 atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg
     1501 actcgattat acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc
     1561 tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga
     1621 ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc
     1681 gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc
     1741 ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagcct aggcaccggg
     1801 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     1861 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     1921 caccccggcg cgctcggcgg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     1981 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     2041 gtgcggcgcc aggaggccct ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     2101 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     2161 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     2221 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac
     2281 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     2341 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgt tgtctaacaa
     2401 aaaagccaaa aacggccaga atttagcgga caatttacaa gtctaacact gaaaattaca
     2461 tattgaccca aatgattaca tttcaaaagg tgcctaaaaa acttcacaaa acacactcgc
     2521 caaccccgag cgcatagttc aaaaccggag cttcagctac ttaagaagat aggtacataa
     2581 aaccgaccaa agaaactgac gcctcactta tccctcccct caccagaggt ccggcgcctg
     2641 tcgattcagg agagcctacc ctaggcccga accctgcgtc ctgcgacgga gaaaagccta
     2701 ccgcacacct accggctggt ggccccaccc tgcattataa gccaacagaa cgggtgacgt
     2761 cacgacacga cgagggcgcg cgctcccaaa ggtacgggtg cactgcccaa cggcaccgcc
     2821 ataactgccg cccccgcaac agacgacaaa ccgagttctc cagtcagtga caaacttcac
     2881 gtcagggtcc ccagatggtg cccagcccat ctcacccgaa taagagcttt cccgcattag
     2941 cgaagggctc aagaccttgg gttcttgccg cccaccatgc cccccacctt gtttcaacga
     3001 cctcacagcc cgcctcacaa gcgtcttcca ttcaagactc gggaacagcc gccattttgc
     3061 tgcgctcccc ccaaccccca gttcagggca accttgctcg cggacccaga ctacagccct
     3121 tggcggtctc tccacacgct tccgtcccac cgagcggccc ggcggccacg aaagccccgg
     3181 ccagcccagc agcccgctac tcaccaagtg acgatcacag cgatccacaa acaagaaccg
     3241 cgacccaaat cccggctgcg acggaactag ctgtgccaca cccggcgcgt ccttatataa
     3301 tcatcggcgt tcaccgcccc acggagatcc ctccgcagaa tcgccgagaa gggactactt
     3361 ttcctcgcct gttccgctct ctggaaagaa aaccagtgcc ctagagtcac ccaagtcccg
     3421 tcctaaaatg tccttctgct gatactgggg ttctaaggcc gagtcttatg agcagcgggc
     3481 cgctgtcctg agcgtccggg cggaaggatc aggacgctcc tgcgcccttc gtctgacgtg
     3541 gcagcgctcg ccgtgacgag gggggcgccc gcgggaggcg ccaaaacccg gcgcggaggc
     3601 caggcgcgcc ccgatttcga tattcctcgg atctcggata gatcgggatt gaggatgtag
     3661 gttgaacttc tctcgggtga tcccgatctc ggatattcgg gattggagta atacctggcg
     3721 ataggcgctg ccaagcaata gccaaggcgc aaggccggcc aagagacttg cgatagttcg
     3781 ttcgagaggt agcctgcaag atggatcgga cgaaaggttt gaagattttc cgacgagggc
     3841 cttggagagt tcggaaattt aaatgcaact atggatgaac gaaatagaca gatcgctgag
     3901 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4741 gctcacatga tttaaatgaa actcgtaatc ggagaacggg tgcaatagca aggaccagat
     4801 cgaacttgac tgacgcaatt ctgagcgtag tcgaaaccca gacaagtcga tctcgccaag
     4861 tagctgactt gactggttaa ttaagtttcg atagcccaag gtaaccaatg agattaggga
     4921 gggaagaaag tttgatgaac tacttctgac ccctagtggt gtccagaaaa gaccattaaa
     4981 ggaatgtctt gatcgaaatc gagagatcgc ctgcaggata gctgacgacc ttaaggataa
     5041 atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt atccgttgct
     5101 gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt tttttctgcg
     5161 ccaatgccgg tagatagcaa tccggcccga ggggcacaaa aaacccctca agacccgttt
     5221 agaggcccca aggggttatg ctacgatacc gatagagatg ggccaatacc acatttgtag
     5281 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5341 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5401 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5461 aactcatcaa tgtatcttat catgtctgtg agctgaaggt acgctgtatc tcagtcagtc
     5521 aagctagctt aggtagaatc taagccgaga agtggattgg gtataggttt ccccgactga
     5581 aagtaaaggt tttctgtgag attaaagtta actacatcta cttgtgctat gtagttacga
     5641 gaattcattc tgcacaagag tagactatat atcgtaaacg gaaaagcgaa aacgtttata
     5701 tagcccatgg tggcggccgc tggcgggtgt gtcgagtgga tggtaggatc gacaaagatc
     5761 tgatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     5821 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     5881 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     5941 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcagcatg gtaatagcga
     6001 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     6061 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     6121 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     6181 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     6241 ggtcgttggg cggtcagcca ggcgggccat tt
//
