LOCUS       sequence-304474-        4878 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 4878)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4878)
  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-m4133684
FEATURES             Location/Qualifiers
     source          1..4878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    67..138
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(143..189)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    207..255
                     /note="synthetic polyadenylation signal"
     enhancer        336..639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        641..843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     794..814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      912..921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1032..1052
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1086..1163
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1191..1211
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1660..1781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1671..1690)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1725..1744
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1896..1942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1972..2108
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2153..2224
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2450..3038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2530..2549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3784..3852,3853..4644)
                     /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(4002..4021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4657..4793
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
       61 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
      121 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
      181 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
      241 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
      301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      601 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
      661 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      721 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      781 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      841 gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc
      901 cgccagcggc cgccaccatg ggctatataa acgttttcgc ttttccgttt acgatatata
      961 gtctactctt gtgcagaatg aattctcgta actacatagc acaagtagat gtagttaact
     1021 ttaatctcac agaaaacctt tactttcagt cgggacggcg aatcccgggg ctgataaatc
     1081 cctggaaaag gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa
     1141 agatcagttc aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct
     1201 atttccaggg tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata
     1261 aagaacagca gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac
     1321 gcaacgcttt cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg
     1381 aagccaagaa gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc
     1441 agcagaatgc tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg
     1501 catttatcca gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca
     1561 aaaagctcaa tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga
     1621 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
     1681 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1741 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1801 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1861 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
     1921 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
     1981 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     2041 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     2101 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     2161 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     2221 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     2281 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     2341 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     2401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2881 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3241 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     3301 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     3361 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     3421 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     3481 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt
     3541 gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca
     3601 gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc
     3661 tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg
     3721 tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt
     3781 cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3841 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3901 gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3961 cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4021 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4081 ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4141 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4201 attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4261 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4321 atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta
     4381 gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg accacttctg
     4441 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4501 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4561 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4621 gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct
     4681 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     4741 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg
     4801 aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca
     4861 gccgtggcgc tcatttgc
//
