LOCUS       sequence-295901-        5589 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 5589)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5589)
  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-m4133754
FEATURES             Location/Qualifiers
     source          1..5589
                     /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             884..961
                     /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             989..1009
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1458..1579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1469..1488)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1523..1542
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1694..1740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1770..1906
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1951..2022
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2248..2836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2328..2347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3582..3650,3651..4442)
                     /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(3800..3819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4455..4591
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4743..4814
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4819..4865)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4883..4931
                     /note="synthetic polyadenylation signal"
     enhancer        5012..5315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5317..5519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5470..5490
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5588..5589,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggat ttgtttatga gaatcttcac aattggaact gtaactttga agcaaggtga
       61 aatcaaggat gctactcctt cagattttgt tcgcgctact gcaacgatac cgatacaagc
      121 ctcactccct ttcggatggc ttattgttgg cgttgcactt cttgctgttt ttcagagcgc
      181 ttccaaaatc ataaccctca aaaagagatg gcaactagca ctctccaagg gtgttcactt
      241 tgtttgcaac ttgctgttgt tgtttgtaac agtttactca caccttttgc tcgttgctgc
      301 tggccttgaa gccccttttc tctatcttta tgctttagtc tacttcttgc agagtataaa
      361 ctttgtaaga ataataatga ggctttggct ttgctggaaa tgccgttcca aaaacccatt
      421 actttatgat gccaactatt ttctttgctg gcatactaat tgttacgact attgtatacc
      481 ttacaatagt gtaacttctt caattgtcat tacttcaggt gatggcacaa caagtcctat
      541 ttctgaacat gactaccaga ttggtggtta tactgaaaaa tgggaatctg gagtaaaaga
      601 ctgtgttgta ttacacagtt acttcacttc agactattac cagctgtact caactcaatt
      661 gagtacagac actggtgttg aacatgttac cttcttcatc tacaataaaa ttgttgatga
      721 gcctgaagaa catgtccaaa ttcacacaat cgacggttca tccggagttg ttaatccagt
      781 aatggaacca atttatgatg aaccgacgac gactactagc gtgcctttgg aaaaccttta
      841 ctttcagtcg ggacggcgaa tcccggggct gataaatccc tggaaaagga gatggaagaa
      901 gaactttatt gcagtgtctg cggctaaccg gttcaaaaag atcagttcaa gcggcgccct
      961 agattacgac attcctacga ccgcctccga gaatctctat ttccagggtg aatttggctt
     1021 ggcacaacat gacgaggccg tcgataacaa gtttaataaa gaacagcaga acgctttcta
     1081 tgagatcctt cacttaccaa atctgaacga agagcaacgc aacgctttca ttcagagcct
     1141 gaaagacgat ccctcgcagt ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc
     1201 acaggctcca aaagttgata acaagtttaa caaggagcag cagaatgctt tctacgagat
     1261 attgcacctt cctaatctga atgaggaaca aagaaatgca tttatccagt ccctaaaaga
     1321 cgatccatca cagagtgcca acctgcttgc ggaagccaaa aagctcaatg atgcccaggc
     1381 cccgaaagta gacgcgaacc atcaataagc tagcttgact gactgagata cagcgtacct
     1441 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1501 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1561 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1621 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
     1681 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1741 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
     1801 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     1861 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
     1921 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     1981 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     2041 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     2101 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     2161 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     2221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3061 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     3121 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     3181 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     3241 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     3301 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag
     3361 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     3421 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     3481 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     3541 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     3601 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3661 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     3721 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     3781 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3841 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     3901 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3961 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     4021 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4081 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4141 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     4201 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4261 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4321 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4381 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4441 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     4501 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     4561 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     4621 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     4681 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     4741 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     4801 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     4861 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     4921 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     4981 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     5041 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     5101 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     5161 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     5221 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     5281 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     5341 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     5401 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5461 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     5521 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     5581 agcggccgc
//
