LOCUS       sequence-345633-        7019 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 7019)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7019)
  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-m4134056
FEATURES             Location/Qualifiers
     source          1..7019
                     /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..1506
                     /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    1559..1680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1570..1589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1624..1643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1795..1841
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1871..2007
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2052..2123
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2349..2937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2429..2448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3437..4646
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4483..4502
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4650..5249
                     /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(4650..4669)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5146..5166
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5276..5324
                     /note="synthetic polyadenylation signal"
     CDS             join(5590..5658,5659..6450)
                     /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(5808..5827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6463..6599
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6751..6822
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6827..6873)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6891..6939
                     /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 ttcagtcgaa gaggcgatgg
     1441 aaaaagaatt ttatcgcggt gagtgcagcc aacagattca aaaagatatc ctcgtctggt
     1501 gccctctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
     1561 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1621 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1681 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1741 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
     1801 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
     1861 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     1921 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     1981 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
     2041 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
     2101 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
     2161 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
     2221 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     2281 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2761 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     3181 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     3241 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     3301 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     3361 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     3421 aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     3481 gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc
     3541 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     3601 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     3661 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     3721 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     3781 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     3841 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     3901 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     3961 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     4021 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     4081 aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac
     4141 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     4201 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     4261 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     4321 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     4381 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     4441 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     4501 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     4561 aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct
     4621 ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg gtgcgcctcg
     4681 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4741 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4801 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4861 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4921 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggagg
     4981 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5041 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggccg
     5101 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5161 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5221 ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct tagataataa
     5281 aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg
     5341 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     5401 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     5461 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     5521 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     5581 ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5641 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5701 ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5761 tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5821 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5881 aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5941 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6001 acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6061 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6121 accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt
     6181 actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca
     6241 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6301 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6361 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6421 ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa
     6481 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     6541 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     6601 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
     6661 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
     6721 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
     6781 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
     6841 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
     6901 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
     6961 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
