LOCUS       sequence-295289-        5112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5112)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5112)
  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-m4134350
FEATURES             Location/Qualifiers
     source          1..5112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             830..850
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             851..928
                     /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    981..1102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(992..1011)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1046..1065
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1217..1263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1293..1429
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1474..1545
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1771..2359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1851..1870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3105..3173,3174..3965)
                     /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(3323..3342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3978..4114
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4266..4337
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4342..4388)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4406..4454
                     /note="synthetic polyadenylation signal"
     enhancer        4535..4838
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4840..5042
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4993..5013
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5111..5112,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggat ctgtttatgc gtatattcac catcggcacg gttacattga agcaagggga
       61 aattaaagac gcaactccca gtgacttcgt acgggcgacc gctacaattc cgatccaggc
      121 cagcctccct tttggatggc ttatcgtcgg cgtggcccta ctggctgtgt ttcagtcggc
      181 atcaaagata atcactctga aaaagcgatg gcagttagcc cttagcaaag gtgtgcactt
      241 cgtatgcaat ttgctgctcc tgttcgtcac cgtttattcc catctgctcc tggtggctgc
      301 cggattggag gcaccatttt tatacctata cgccctcgtg tattttctgc aatctattaa
      361 cttcgtccgc attatcatga gactctggct ctgttggaag tgcaggtcta aaaatccctt
      421 gttatatgat gccaactact tcctgtgttg gcacactaac tgctatgact actgtattcc
      481 atacaattca gttacctcca gtatagtgat cacatctggg gatggtacga cttcgcctat
      541 ttccgagcat gactatcaga tcggcgggta cacagaaaag tgggagagcg gagtcaagga
      601 ttgcgtggtg cttcactcct atttcaccag tgactactac cagctttata gcacacaact
      661 ttcaacagac accggcgtgg agcatgtgac ttttttcatt tataataaaa tcgttgatga
      721 accggaggaa cacgttcaga tacacactat cgacggcagc tctggtgtcg tcaaccctgt
      781 tatggagccc atttacgatg aaccaacgac cacaacatct gtacctctgg aaaaccttta
      841 ctttcagtcg aagaggcgat ggaaaaagaa ttttatcgcg gtgagtgcag ccaacagatt
      901 caaaaagata tcctcgtctg gtgccctcta agctagcttg actgactgag atacagcgta
      961 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1021 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1081 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1141 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     1201 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1261 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     1321 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     1381 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     1441 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     1501 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1561 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     1621 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     1681 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1741 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1801 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1861 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1921 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1981 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2041 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2101 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2161 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2221 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2281 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2341 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2401 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2461 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2521 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2581 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     2641 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     2701 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2761 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2821 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     2881 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     2941 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     3001 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     3061 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     3121 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3181 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     3241 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     3301 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3361 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     3421 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3481 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     3541 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3601 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3661 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     3721 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3781 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3841 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3901 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3961 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     4021 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     4081 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     4141 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     4201 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     4261 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     4321 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     4381 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     4441 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     4501 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4561 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4621 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4681 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4741 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4801 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     4861 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4921 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4981 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5041 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     5101 gccagcggcc gc
//
