LOCUS       sequence-311980-        5079 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 5079)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5079)
  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-m4134531
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1408..1428
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1429..1473
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1526..1647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1537..1556)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1591..1610
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1762..1808
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1838..1974
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2019..2090
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2316..2904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2396..2415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3650..3718,3719..4510)
                     /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(3868..3887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4523..4659
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4811..4882
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4887..4933)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4951..4999
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggatct gtttatgcgt
      601 atattcacca tcggcacggt tacattgaag caaggggaaa ttaaagacgc aactcccagt
      661 gacttcgtac gggcgaccgc tacaattccg atccaggcca gcctcccttt tggatggctt
      721 atcgtcggcg tggccctact ggctgtgttt cagtcggcat caaagataat cactctgaaa
      781 aagcgatggc agttagccct tagcaaaggt gtgcacttcg tatgcaattt gctgctcctg
      841 ttcgtcaccg tttattccca tctgctcctg gtggctgccg gattggaggc accattttta
      901 tacctatacg ccctcgtgta ttttctgcaa tctattaact tcgtccgcat tatcatgaga
      961 ctctggctct gttggaagtg caggtctaaa aatcccttgt tatatgatgc caactacttc
     1021 ctgtgttggc acactaactg ctatgactac tgtattccat acaattcagt tacctccagt
     1081 atagtgatca catctgggga tggtacgact tcgcctattt ccgagcatga ctatcagatc
     1141 ggcgggtaca cagaaaagtg ggagagcgga gtcaaggatt gcgtggtgct tcactcctat
     1201 ttcaccagtg actactacca gctttatagc acacaacttt caacagacac cggcgtggag
     1261 catgtgactt ttttcattta taataaaatc gttgatgaac cggaggaaca cgttcagata
     1321 cacactatcg acggcagctc tggtgtcgtc aaccctgtta tggagcccat ttacgatgaa
     1381 ccaacgacca caacatctgt acctctggaa aacctttact ttcagtcgaa agaaaccgcg
     1441 gcagctaagt tcgagaggca gcacatggac agttaagcta gcttgactga ctgagataca
     1501 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     1561 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1621 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1681 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     1741 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     1801 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     1861 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     1921 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     1981 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     2041 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     2101 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     2161 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     2221 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     2281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3121 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     3181 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     3241 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     3301 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     3361 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat
     3421 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     3481 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     3541 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     3601 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     3661 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3721 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     3781 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     3841 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3901 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     3961 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4021 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4081 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4141 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4201 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4261 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4321 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4381 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4441 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4501 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     4561 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     4621 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     4681 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     4741 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     4801 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     4861 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     4921 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     4981 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5041 tacggggtca ttagttcata gcccatatat ggagttccg
//
