LOCUS       sequence-300428-        6320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6320)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6320)
  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-m4252979
FEATURES             Location/Qualifiers
     source          1..6320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    131..202
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(207..253)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    271..319
                     /note="synthetic polyadenylation signal"
     enhancer        400..703
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        705..907
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     858..878
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1259..1380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1270..1289)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1324..1343
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1495..1541
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1571..1707
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1752..1823
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2049..2637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2129..2148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3137..4346
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4183..4202
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4350..4949
                     /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(4350..4369)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4846..4866
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4976..5024
                     /note="synthetic polyadenylation signal"
     CDS             join(5290..5358,5359..6150)
                     /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(5508..5527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6163..6299
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
       61 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
      121 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
      181 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
      241 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
      301 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
      361 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      421 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      481 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      541 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      601 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      661 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
      721 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      781 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      841 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      901 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
      961 cacccgccag cggccgccac catgtacagc tttgtatctg aagagacagg cactctcatc
     1021 gttaattcag tgttgctgtt cctggcattt gtggtcttcc ttttagtgac cctagcgata
     1081 ctgacggccc tccggctgtg cgcttattgt tgtaacattg tcaacgttag tctcgtgaag
     1141 ccctcgttct acgtttattc ccgcgtgaaa aatctaaaca gctcccgtgt gcctgatctt
     1201 cttgtctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
     1261 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1321 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1381 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1441 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
     1501 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
     1561 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     1621 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     1681 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
     1741 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
     1801 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
     1861 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
     1921 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     1981 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2221 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2461 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2641 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2701 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     2881 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     2941 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     3001 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     3061 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     3121 aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     3181 gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc
     3241 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     3301 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     3361 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     3421 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     3481 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     3541 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     3601 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     3661 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     3721 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     3781 aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac
     3841 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     3901 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     3961 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     4021 agccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     4081 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     4141 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     4201 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     4261 aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct
     4321 ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg gtgcgcctcg
     4381 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4441 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4501 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4561 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4621 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggagg
     4681 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4741 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggccg
     4801 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4861 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4921 ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct tagataataa
     4981 aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg
     5041 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     5101 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     5161 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     5221 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     5281 ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5341 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5401 ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5461 tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5521 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5581 aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5641 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5701 acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5761 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5821 accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt
     5881 actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca
     5941 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6001 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6061 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6121 ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa
     6181 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     6241 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     6301 atcccgaatc gtttaaactc
//
