LOCUS       sequence-300753-        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"
     misc_feature    8..79
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(231..367)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(380..1171,1172..1240))
                     /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     1003..1022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1986..2574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2475..2494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2800..2871
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2916..3052)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3082..3128)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3243..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3334..3353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3417..3494)
                     /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             complement(3495..3515)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      3626..3635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3704..3906)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3733..3753)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3908..4211
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4292..4340
                     /note="synthetic polyadenylation signal"
     terminator      join(4358..4401,1..3)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttgaattgag aggtagctga agctgctgtt atgaccacta gagggaagaa gatacctgtg
       61 gagctaatgg tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa gctgacgata
      121 gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc gtaagctcga
      181 aacgacggag attcgataga gccagagtcg agtttaaacg attcgggatc ttaaggataa
      241 atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt atccgttgct
      301 gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt tttttctgcg
      361 ccaatgcacc tagaatcggt taccaatgct taatcagtga ggcacctatc tcagcgatct
      421 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      481 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      541 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
      601 ctttatccgc ctccatccag tctatcaact gttgccggga agctagagta agtagttcgc
      661 cagttaatag tttacgcaag gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
      721 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      781 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc agaagtaagt
      841 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
      901 catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt
      961 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1021 gcagaacttt aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa ctctcaagga
     1081 tcttaccgct gttgagatcc agttcgatgt aacccactcg cgcacccaac tgatcttcag
     1141 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1201 aaaagggaat aagggcgaca cggaaatgtt gaatactcat gcgaaagcct cctgctgatc
     1261 ctgtctcttg atagatgttt gcaaaagcta ggactcgatt atacgttgac cgatcagact
     1321 atcaatatct aggtttaaac agatcggtac gctgccgtga ccaggcttga gtacaatatc
     1381 tgacgattgg acttcctggg tcccaagctg gattccgact ggctgactgc caagcaacga
     1441 tatggatcgt cgagactcgc actgagagga tcgttctcgg tacaccaggc gcgccccgat
     1501 ttcgatattc ctcggatctc ggatagatcg ggattgagga tgtaggttga acttctctcg
     1561 ggtgatcccg atctcggata ttcgggattg gagtaatacc tggcgatagg cgctgccaag
     1621 caatagccaa ggcgcaaggc cggccaagag acttgcgata gttcgttcga gaggtagcct
     1681 gcaagatgga tcggacgaaa ggtttgaaga ttttccgacg agggccttgg agagttcgga
     1741 aatttaaatg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1801 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2101 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2281 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgatttaa
     2641 atgaaactcg taatcggaga acgggtgcaa tagcaaggac cagatcgaac ttgactgacg
     2701 caattctgag cgtagtcgaa acccagacaa gtcgatctcg ccaagtagct gacttgactg
     2761 gttaattaag tttcgatagc ccaaggtaac caatgagatt agggagggaa gaaagtttga
     2821 tgaactactt ctgaccccta gtggtgtcca gaaaagacca ttaaaggaat gtcttgatcg
     2881 aaatcgagag atcgcctgca ggatagctga cgaccttaag gataaatttc tggtaaggag
     2941 gacacgtatg gaagtgggca agttggggaa gccgtatccg ttgctgaatc tggcatatgt
     3001 gggagtataa gacgcgcagc gtcgcatcag gcattttttt ctgcgccaat gccggtagat
     3061 agcaatccgg cccgaggggc acaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     3121 ttatgctacg ataccgatag agatgggcca ataccacatt tgtagaggtt ttacttgctt
     3181 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3241 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3361 cttatcatgt ctgtgagctg aaggtacgct gtatctcagt cagtcaagct agcttagagg
     3421 gcaccagacg aggatatctt tttgaatctg ttggctgcac tcaccgcgat aaaattcttt
     3481 ttccatcgcc tcttcgactg aaagtaaagg ttttctgtga gattaaagtt aactacatct
     3541 acttgtgcta tgtagttacg agaattcatt ctgcacaaga gtagactata tatcgtaaac
     3601 ggaaaagcga aaacgtttat atagcccatg gtggcggccg ctggcgggtg tgtcgagtgg
     3661 atggtaggat cgacaaagat ctgatctgac ggttcactaa accagctctg cttatataga
     3721 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     3781 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     3841 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     3901 gcatcagcat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3961 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4021 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4081 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4141 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     4201 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     4261 agatctggct acactcgatc agcagttaga taataaaatc gctatccatc gaagatggat
     4321 gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag cataacccct tggggcctct
     4381 aaacgggtct tgaggggttt t
//
