LOCUS       sequence-300690-        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             complement(79..96)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(97..117)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      228..237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(306..508)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(335..355)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        510..813
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    894..942
                     /note="synthetic polyadenylation signal"
     terminator      960..1006
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1011..1082
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1234..1370)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1383..2174,2175..2243))
                     /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     2006..2025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2989..3577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3478..3497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3803..3874
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3919..4055)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(4085..4131)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(4246..4341,1..26)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4283..4302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     join(4337..4341,1..15)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttgtccaaac tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca
       61 gtcagtcaag ctagcttaat ggtgatggtg atggtgcgac tgaaagtaaa ggttttctgt
      121 gagattaaag ttaactacat ctacttgtgc tatgtagtta cgagaattca ttctgcacaa
      181 gagtagacta tatatcgtaa acggaaaagc gaaaacgttt atatagccca tggtggcggc
      241 cgctggcggg tgtgtcgagt ggatggtagg atcgacaaag atctgatctg acggttcact
      301 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
      361 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
      421 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
      481 ccattgatgt actgccaaaa ccgcatcagc atggtaatag cgatgactaa tacgtagatg
      541 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
      601 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
      661 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
      721 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
      781 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
      841 aatgaccccg taattgatta ctagatctgg ctacactcga tcagcagtta gataataaaa
      901 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat
      961 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgaatt gagaggtagc
     1021 tgaagctgct gttatgacca ctagagggaa gaagatacct gtggagctaa tggtccaaga
     1081 tgtcgggagc cgcgatcgct gccaaaaagg taagctgacg atagctgacg agattcgatg
     1141 cccgacgagc aaatgagcgc cacggctgtt cgcgtaagct cgaaacgacg gagattcgat
     1201 agagccagag tcgagtttaa acgattcggg atcttaagga taaatttctg gtaaggagga
     1261 cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg
     1321 gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc acctagaatc
     1381 ggttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1441 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1501 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1561 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1621 cagtctatca actgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttacgc
     1681 aaggttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1741 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1801 gcggttagct ccttcggtcc tccaatcgtt gtcagaagta agttggccgc agtgttatca
     1861 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1921 tctgtgactg gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1981 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2041 ctcatcattg gaaagcgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2101 tccagttcga tgtaacccac tcgcgcaccc aactgatctt cagcatcttt tactttcacc
     2161 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2221 acacggaaat gttgaatact catgcgaaag cctcctgctg atcctgtctc ttgatagatg
     2281 tttgcaaaag ctaggactcg attatacgtt gaccgatcag actatcaata tctaggttta
     2341 aacagatcgg tacgctgccg tgaccaggct tgagtacaat atctgacgat tggacttcct
     2401 gggtcccaag ctggattccg actggctgac tgccaagcaa cgatatggat cgtcgagact
     2461 cgcactgaga ggatcgttct cggtacacca ggcgcgcccc gatttcgata ttcctcggat
     2521 ctcggataga tcgggattga ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg
     2581 atattcggga ttggagtaat acctggcgat aggcgctgcc aagcaatagc caaggcgcaa
     2641 ggccggccaa gagacttgcg atagttcgtt cgagaggtag cctgcaagat ggatcggacg
     2701 aaaggtttga agattttccg acgagggcct tggagagttc ggaaatttaa atgcaactat
     2761 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2821 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3121 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3601 acggttcctg gccttttgct ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg
     3661 agaacgggtg caatagcaag gaccagatcg aacttgactg acgcaattct gagcgtagtc
     3721 gaaacccaga caagtcgatc tcgccaagta gctgacttga ctggttaatt aagtttcgat
     3781 agcccaaggt aaccaatgag attagggagg gaagaaagtt tgatgaacta cttctgaccc
     3841 ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga tcgaaatcga gagatcgcct
     3901 gcaggatagc tgacgacctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
     3961 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
     4021 agcgtcgcat caggcatttt tttctgcgcc aatgccggta gatagcaatc cggcccgagg
     4081 ggcacaaaaa acccctcaag acccgtttag aggccccaag gggttatgct acgataccga
     4141 tagagatggg ccaataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4201 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4261 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4321 tcactgcatt ctagttgtgg t
//
