LOCUS       sequence-297431-        4401 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 4401)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4401)
  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-m4133837
FEATURES             Location/Qualifiers
     source          1..4401
                     /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..217
                     /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"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      506..552
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      582..718
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    763..834
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1060..1648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1140..1159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2394..2462,2463..3254)
                     /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(2612..2631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3267..3403
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3555..3626
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3631..3677)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3695..3743
                     /note="synthetic polyadenylation signal"
     enhancer        3824..4127
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4129..4331
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4282..4302
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4400..4401,1..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 tttcagtcga agaggcgatg gaaaaagaat tttatcgcgg tgagtgcagc
      181 caacagattc aaaaagatat cctcgtctgg tgccctctaa gctagcttga ctgactgaga
      241 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      421 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      481 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
      541 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
      601 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
      661 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
      721 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
      781 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
      841 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
      901 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
      961 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1861 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     1921 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     1981 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2041 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2101 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2161 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2221 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     2281 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     2341 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     2401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     2521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2581 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2701 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     2821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2941 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3001 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3241 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     3301 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     3361 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     3421 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     3481 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     3541 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     3601 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     3661 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     3721 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     3781 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3841 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3901 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     3961 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4021 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4081 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4141 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4201 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4261 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4321 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     4381 gacacacccg ccagcggccg c
//
