LOCUS       sequence-300700-        4452 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 4452)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4452)
  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-m4133244
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(114..131)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(132..152)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(452..654)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(481..501)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        656..959
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1040..1088
                     /note="synthetic polyadenylation signal"
     terminator      1106..1152
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1157..1228
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1380..1516)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1529..2320,2321..2389))
                     /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     2152..2171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3135..3723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3624..3643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3949..4020
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4065..4201)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(4231..4277)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(4392..4452,1..61)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4429..4448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
       61 atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc ttaatggtga
      121 tggtgatggt gcgactgaaa gtaaaggttt tcgaccagaa gatcaggaac tctagaagaa
      181 ttcagatttt taacacgaga gtaaacgtaa aaagaaggtt ttacaagact cacgttaaca
      241 atattgcagc agtacgcaca caatcgaagc gcagtaagga tggctagtgt aactagcaag
      301 aataccacga aagcaagaaa aagaagtacg ctattaacta ttaacgtacc tgtctcttcc
      361 gaaacgaatg agtacatggt ggcggccgct ggcgggtgtg tcgagtggat ggtaggatcg
      421 acaaagatct gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta
      481 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
      541 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
      601 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcagcatgg
      661 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
      721 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
      781 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
      841 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
      901 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
      961 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactag atctggctac
     1021 actcgatcag cagttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
     1081 tttgtgtgtg taacgcaacg attgatagca taaccccttg gggcctctaa acgggtcttg
     1141 aggggttttt tgaattgaga ggtagctgaa gctgctgtta tgaccactag agggaagaag
     1201 atacctgtgg agctaatggt ccaagatgtc gggagccgcg atcgctgcca aaaaggtaag
     1261 ctgacgatag ctgacgagat tcgatgcccg acgagcaaat gagcgccacg gctgttcgcg
     1321 taagctcgaa acgacggaga ttcgatagag ccagagtcga gtttaaacga ttcgggatct
     1381 taaggataaa tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta
     1441 tccgttgctg aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt
     1501 ttttctgcgc caatgcacct agaatcggtt accaatgctt aatcagtgag gcacctatct
     1561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1741 gtcctgcaac tttatccgcc tccatccagt ctatcaactg ttgccgggaa gctagagtaa
     1801 gtagttcgcc agttaatagt ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt
     1861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca
     1981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga atactcaacc aagtcattct
     2101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2161 cgccacatag cagaacttta aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac
     2221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgc gcacccaact
     2281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatg cgaaagcctc
     2401 ctgctgatcc tgtctcttga tagatgtttg caaaagctag gactcgatta tacgttgacc
     2461 gatcagacta tcaatatcta ggtttaaaca gatcggtacg ctgccgtgac caggcttgag
     2521 tacaatatct gacgattgga cttcctgggt cccaagctgg attccgactg gctgactgcc
     2581 aagcaacgat atggatcgtc gagactcgca ctgagaggat cgttctcggt acaccaggcg
     2641 cgccccgatt tcgatattcc tcggatctcg gatagatcgg gattgaggat gtaggttgaa
     2701 cttctctcgg gtgatcccga tctcggatat tcgggattgg agtaatacct ggcgataggc
     2761 gctgccaagc aatagccaag gcgcaaggcc ggccaagaga cttgcgatag ttcgttcgag
     2821 aggtagcctg caagatggat cggacgaaag gtttgaagat tttccgacga gggccttgga
     2881 gagttcggaa atttaaatgc aactatggat gaacgaaata gacagatcgc tgagataggt
     2941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3241 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3781 atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat agcaaggacc agatcgaact
     3841 tgactgacgc aattctgagc gtagtcgaaa cccagacaag tcgatctcgc caagtagctg
     3901 acttgactgg ttaattaagt ttcgatagcc caaggtaacc aatgagatta gggagggaag
     3961 aaagtttgat gaactacttc tgacccctag tggtgtccag aaaagaccat taaaggaatg
     4021 tcttgatcga aatcgagaga tcgcctgcag gatagctgac gaccttaagg ataaatttct
     4081 ggtaaggagg acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct
     4141 ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg
     4201 ccggtagata gcaatccggc ccgaggggca caaaaaaccc ctcaagaccc gtttagaggc
     4261 cccaaggggt tatgctacga taccgataga gatgggccaa taccacattt gtagaggttt
     4321 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4381 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4441 caaatttcac aa
//
