LOCUS       sequence-297574-        6320 bp    DNA     circular SYN 15-APR-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 Apr 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"
     polyA_signal    282..403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(293..312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     347..366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      518..564
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      594..730
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    775..846
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1072..1660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1152..1171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2160..3369
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3206..3225
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3373..3972
                     /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(3373..3392)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3869..3889
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3999..4047
                     /note="synthetic polyadenylation signal"
     CDS             join(4313..4381,4382..5173)
                     /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(4531..4550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5186..5322
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5474..5545
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5550..5596)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5614..5662
                     /note="synthetic polyadenylation signal"
     enhancer        5743..6046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6048..6250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6201..6221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac agctttgtat ctgaagagac aggcactctc atcgttaatt cagtgttgct
       61 gttcctggca tttgtggtct tccttttagt gaccctagcg atactgacgg ccctccggct
      121 gtgcgcttat tgttgtaaca ttgtcaacgt tagtctcgtg aagccctcgt tctacgttta
      181 ttcccgcgtg aaaaatctaa acagctcccg tgtgcctgat cttcttgtct aagctagctt
      241 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      301 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      361 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      421 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      481 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
      541 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
      601 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
      661 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
      721 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
      781 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
      841 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
      901 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
      961 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1861 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     1921 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     1981 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2041 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2101 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2161 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
     2221 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
     2281 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     2341 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     2401 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     2461 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     2521 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
     2581 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     2641 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     2701 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     2761 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
     2821 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
     2881 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     2941 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     3001 acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt aggtgtgcgg
     3061 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     3121 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     3181 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     3241 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     3301 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     3361 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3421 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3481 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3541 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3601 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3661 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac
     3721 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3781 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg
     3841 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3901 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     3961 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa
     4021 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac
     4081 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4141 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4201 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4261 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4321 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4381 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4441 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4501 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4561 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4621 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4681 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     4741 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4801 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4861 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     4921 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4981 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5041 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5101 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5161 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5221 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5281 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa
     5341 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     5401 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     5461 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     5521 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     5581 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     5641 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
     5701 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     5761 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     5821 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     5881 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     5941 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6001 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
     6061 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     6121 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     6181 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     6241 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
     6301 acacacccgc cagcggccgc
//
