LOCUS       sequence-300635-        4365 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 4365)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4365)
  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-m4133675
FEATURES             Location/Qualifiers
     source          1..4365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    29..77
                     /note="synthetic polyadenylation signal"
     terminator      95..141
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    146..217
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(369..505)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(518..1309,1310..1378))
                     /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     1141..1160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2124..2712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2613..2632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2938..3009
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3054..3190)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3220..3266)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3381..3502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3418..3437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3472..3491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3555..3596)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3597..3617)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      3728..3737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3806..4008)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3835..3855)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4010..4313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tctggctaca ctcgatcagc agttagataa taaaatcgct atccatcgaa gatggatgtg
       61 tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg ggcctctaaa
      121 cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat gaccactaga
      181 gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga tcgctgccaa
      241 aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg agcgccacgg
      301 ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag tttaaacgat
      361 tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg
      421 gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat
      481 caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta atcagtgagg
      541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
      661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt tgccgggaag
      781 ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt gctacaggca
      841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccaa
      961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa tactcaacca
     1081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag cgttcttcgg
     1201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgcg
     1261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatgc
     1381 gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg actcgattat
     1441 acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc tgccgtgacc
     1501 aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga ttccgactgg
     1561 ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc gttctcggta
     1621 caccaggcgc gccccgattt cgatattcct cggatctcgg atagatcggg attgaggatg
     1681 taggttgaac ttctctcggg tgatcccgat ctcggatatt cgggattgga gtaatacctg
     1741 gcgataggcg ctgccaagca atagccaagg cgcaaggccg gccaagagac ttgcgatagt
     1801 tcgttcgaga ggtagcctgc aagatggatc ggacgaaagg tttgaagatt ttccgacgag
     1861 ggccttggag agttcggaaa tttaaatgca actatggatg aacgaaatag acagatcgct
     1921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2041 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2701 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2761 tttgctcaca tgatttaaat gaaactcgta atcggagaac gggtgcaata gcaaggacca
     2821 gatcgaactt gactgacgca attctgagcg tagtcgaaac ccagacaagt cgatctcgcc
     2881 aagtagctga cttgactggt taattaagtt tcgatagccc aaggtaacca atgagattag
     2941 ggagggaaga aagtttgatg aactacttct gacccctagt ggtgtccaga aaagaccatt
     3001 aaaggaatgt cttgatcgaa atcgagagat cgcctgcagg atagctgacg accttaagga
     3061 taaatttctg gtaaggagga cacgtatgga agtgggcaag ttggggaagc cgtatccgtt
     3121 gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc atttttttct
     3181 gcgccaatgc cggtagatag caatccggcc cgaggggcac aaaaaacccc tcaagacccg
     3241 tttagaggcc ccaaggggtt atgctacgat accgatagag atgggccaat accacatttg
     3301 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3361 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3421 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3481 ccaaactcat caatgtatct tatcatgtct gtgagctgaa ggtacgctgt atctcagtca
     3541 gtcaagctag cttaggtaga atctaagccg agaagtggat tgggtatagg tttccccgac
     3601 tgaaagtaaa ggttttcagt taggttaaag ttcacgacgt ctacctgagc aatgtaattg
     3661 cgagagttca tcctacataa aaggagtgaa tagatcgtga atggaaaggc gaaaacattt
     3721 atgtacccca tggtggcggc cgctggcggg tgtgtcgagt ggatggtagg atcgacaaag
     3781 atctgatctg acggttcact aaaccagctc tgcttatata gacctcccac cgtacacgcc
     3841 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     3901 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     3961 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcagc atggtaatag
     4021 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4081 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4141 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4201 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4261 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4321 ccatatatgg gctatgaact aatgaccccg taattgatta ctaga
//
