LOCUS       sequence-313650-        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-m4252971
FEATURES             Location/Qualifiers
     source          1..6320
                     /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"
     polyA_signal    860..981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(871..890)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     925..944
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1096..1142
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1172..1308
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1353..1424
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1650..2238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1730..1749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2738..3947
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3784..3803
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3951..4550
                     /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(3951..3970)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4447..4467
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4577..4625
                     /note="synthetic polyadenylation signal"
     CDS             join(4891..4959,4960..5751)
                     /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(5109..5128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5764..5900
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6052..6123
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6128..6174)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6192..6240
                     /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 ccatgtactc attcgtttcg
      601 gaagagacag gtacgttaat agttaatagc gtacttcttt ttcttgcttt cgtggtattc
      661 ttgctagtta cactagccat ccttactgcg cttcgattgt gtgcgtactg ctgcaatatt
      721 gttaacgtga gtcttgtaaa accttctttt tacgtttact ctcgtgttaa aaatctgaat
      781 tcttctagag ttcctgatct tctggtctaa gctagcttga ctgactgaga tacagcgtac
      841 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
      901 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
      961 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1021 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1081 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1141 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     1201 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     1261 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     1321 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     1381 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     1441 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     1501 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     1561 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     1621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2221 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2281 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2341 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2401 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2461 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     2521 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     2581 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     2641 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     2701 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     2761 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     2821 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     2881 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     2941 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     3001 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     3061 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     3121 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     3181 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     3241 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     3301 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     3361 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     3421 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     3481 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     3541 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     3601 ataatgcagg gtggggccac cagccggtag gtgtgcggta ggcttttctc cgtcgcagga
     3661 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     3721 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     3781 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     3841 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     3901 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     3961 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4021 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4081 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4141 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4201 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4261 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4321 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4381 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4441 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4501 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     4561 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     4621 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     4681 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     4741 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     4801 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     4861 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     4921 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4981 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5041 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     5101 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5161 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     5221 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5281 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     5341 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5401 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     5461 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     5521 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5581 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5641 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5701 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     5761 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     5821 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     5881 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     5941 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6001 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6061 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     6121 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     6181 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     6241 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     6301 agcccatata tggagttccg
//
