LOCUS       sequence-310427-        7010 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 7010)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7010)
  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-m4133648
FEATURES             Location/Qualifiers
     source          1..7010
                     /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             1432..1497
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1550..1671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1561..1580)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1615..1634
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1786..1832
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1862..1998
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2043..2114
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2340..2928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2420..2439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3428..4637
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4474..4493
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4641..5240
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4641..4660)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5137..5157
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5267..5315
                     /note="synthetic polyadenylation signal"
     CDS             join(5581..5649,5650..6441)
                     /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(5799..5818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6454..6590
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6742..6813
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6818..6864)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6882..6930
                     /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 ccatggattt gtttatgaga
      601 atcttcacaa ttggaactgt aactttgaag caaggtgaaa tcaaggatgc tactccttca
      661 gattttgttc gcgctactgc aacgataccg atacaagcct cactcccttt cggatggctt
      721 attgttggcg ttgcacttct tgctgttttt cagagcgctt ccaaaatcat aaccctcaaa
      781 aagagatggc aactagcact ctccaagggt gttcactttg tttgcaactt gctgttgttg
      841 tttgtaacag tttactcaca ccttttgctc gttgctgctg gccttgaagc cccttttctc
      901 tatctttatg ctttagtcta cttcttgcag agtataaact ttgtaagaat aataatgagg
      961 ctttggcttt gctggaaatg ccgttccaaa aacccattac tttatgatgc caactatttt
     1021 ctttgctggc atactaattg ttacgactat tgtatacctt acaatagtgt aacttcttca
     1081 attgtcatta cttcaggtga tggcacaaca agtcctattt ctgaacatga ctaccagatt
     1141 ggtggttata ctgaaaaatg ggaatctgga gtaaaagact gtgttgtatt acacagttac
     1201 ttcacttcag actattacca gctgtactca actcaattga gtacagacac tggtgttgaa
     1261 catgttacct tcttcatcta caataaaatt gttgatgagc ctgaagaaca tgtccaaatt
     1321 cacacaatcg acggttcatc cggagttgtt aatccagtaa tggaaccaat ttatgatgaa
     1381 ccgacgacga ctactagcgt gcctttggaa aacctttact ttcagtcgat ggattataag
     1441 gaccacgatg gagactacaa agatcatgac attgattaca aggacgacga tgacaaataa
     1501 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     1561 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1621 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1681 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     1741 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     1801 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     1861 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     1921 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     1981 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     2041 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     2101 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     2161 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     2221 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     2281 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2341 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2401 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2461 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2521 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2581 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2641 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2701 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2761 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2821 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2881 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2941 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3001 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3061 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3121 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     3181 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     3241 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     3301 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     3361 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     3421 cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg
     3481 agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca
     3541 ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca
     3601 ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag
     3661 gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac
     3721 gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat
     3781 ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc
     3841 tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga
     3901 gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg
     3961 ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg
     4021 ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga
     4081 gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg
     4141 accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg
     4201 gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc
     4261 gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag
     4321 gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc
     4381 tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt
     4441 ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct
     4501 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg
     4561 ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt
     4621 tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4681 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4741 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4801 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4861 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4921 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag ggcctcctgg
     4981 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5041 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg
     5101 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5161 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5221 cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata aaatcgctat
     5281 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta
     5341 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc
     5401 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg
     5461 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat
     5521 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc
     5581 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5641 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg
     5701 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5761 gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5821 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5881 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5941 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt
     6001 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6061 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6121 cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct
     6181 tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6241 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6301 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6361 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6421 tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat
     6481 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     6541 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat
     6601 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     6661 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     6721 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     6781 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     6841 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     6901 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     6961 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
