LOCUS       sequence-307341-        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-m4133928
FEATURES             Location/Qualifiers
     source          1..4365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             119..139
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             140..181
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    234..355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(245..264)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     299..318
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      470..516
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      546..682
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    727..798
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1024..1612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1104..1123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2358..2426,2427..3218)
                     /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(2576..2595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3231..3367
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3519..3590
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3595..3641)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3659..3707
                     /note="synthetic polyadenylation signal"
     enhancer        3788..4091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4093..4295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4246..4266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4364..4365,1..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 atctcacaga
      121 aaacctttac tttcagtcgg ggaaacctat acccaatcca cttctcggct tagattctac
      181 ctaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
      241 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      301 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      361 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      421 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
      481 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
      541 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
      601 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
      661 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
      721 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
      781 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
      841 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
      901 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
      961 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1441 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1741 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1801 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     1861 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     1921 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     1981 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     2041 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     2101 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     2161 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     2221 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     2281 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     2341 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2401 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2461 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2521 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     2581 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2641 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     2701 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2761 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2821 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2881 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     2941 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     3001 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3061 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3121 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3181 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     3241 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     3301 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     3361 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     3421 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     3481 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     3541 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     3601 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     3661 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     3721 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     3781 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     3841 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     3901 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     3961 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4021 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4081 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4141 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4201 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4261 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     4321 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
