LOCUS       sequence-313669-        4385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4385)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4385)
  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-m4136453
FEATURES             Location/Qualifiers
     source          1..4385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             122..772
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             779..796
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             827..847
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1220..1251
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1252..1354
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(1355..1423,1424..2215)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1573..1592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2216..2407
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      2431..2458
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2470..3057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2550..2569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3079..3108)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(3194..4276)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat gcaccatcat caccatcatg gtggttctgg
      121 ttcgccgata cttggttatt ggaagattaa aggactggtt cagcccacac gcctcctgct
      181 agaatactta gaggaaaaat atgaagagca cctgtacgaa cgtgatgagg gggacaaatg
      241 gcgaaacaag aaatttgaat tgggcttaga atttccaaat ttgccgtatt atatcgatgg
      301 cgacgtaaaa ctgacccaat caatggccat tattcgctac atcgcagata aacataacat
      361 gctgggtggc tgccctaaag aacgggctga gatctccatg ctggaaggtg cggtgctcga
      421 cattcgctac ggcgtcagta gaattgcgta tagcaaggat ttcgaaactt taaaagtgga
      481 tttcctgagc aaacttccgg agatgctgaa aatgtttgag gataggcttt gtcataagac
      541 gtatctcaat ggtgatcatg ttacccaccc ggactttatg ctgtacgacg cccttgatgt
      601 cgttctgtat atggatccca tgtgcctgga cgccttcccg aaattagtgt gttttaaaaa
      661 acgtatcgaa gcgattccac agatcgataa atacttgaag tcttctaaat atattgcttg
      721 gcctctgcaa ggctggcagg caacgttcgg tgggggagat catccgccaa aatcagatct
      781 ggtgccgcgt gggagccctg gcatacaccg cgacggtggt tctggtgaga acctgtactt
      841 tcaaagctat agttttgtat cagaggaaac ggggacactt attgtcaata gcgtgttgct
      901 gttcctagct ttcgttgttt ttctgctcgt gaccttagcg atcctgactg ccttgcgtct
      961 gtgtgcatac tgctgcaaca tcgttaacgt atccctggta aagccgtctt tctacgtgta
     1021 ttcgcgcgtt aaaaacctca acagctctag ggttcccgac ctgcttgttt aaagagaata
     1081 taaaaagcca gattattaat ccggcttttt tattatttag gttctgttaa gtaactgaac
     1141 ccaatgtcgt tagtgacgct tacctcttaa gaggtcactg acctaacagc ttgactccag
     1201 cgtaactgga ctgcaatcaa ctcaaaagcc tccgaccgga ggcttttgac ttgatcggca
     1261 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     1321 gttatcgaga ttttcaggag ctaaggaagc taaaatgagt attcaacatt tccgtgtcgc
     1381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1501 caacagcggt aagatccttg agagtttacg ccccgaagaa cgttttccaa tgatgagcac
     1561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1681 gcatctcacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1741 taacactgcg gccaacttac ttctggcaac gatcggagga ccgaaggagc taaccgcttt
     1801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1981 ggaggcggat aaagttgcag gatcacttct gcgctcggcc ctcccggctg gctggtttat
     2041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2101 agatggtaag ccctcccgca tcgtagttat ctacacgacg gggagtcagg caactatgga
     2161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaagtgac
     2221 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     2281 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     2341 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     2401 cctctgatga gggcccaaat gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt
     2461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gatgctcaag
     2521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2941 gccagttacc tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3001 agcggtggtc tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3061 gatcctttga ttttctaccg aagaaaggcc cacccgtgaa ggtgagccag tgactttctg
     3121 ctatggaggt caggtatgat ttaaatggtc agtgcggcgc gccaacaaaa aacccctagc
     3181 cccccgtttt atatcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3241 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     3301 ttttcaccag tgagactggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     3361 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     3421 acggcgggat ataacatgag ctatcttcgg tatcgtcgta tcccactacc gagatatccg
     3481 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     3541 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     3601 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     3661 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     3721 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     3781 cgtcctcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     3841 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     3901 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     3961 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     4021 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     4081 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     4141 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     4201 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     4261 acgttactgg tttcatattc accaccctga attgactgtc ttccgggcgc tatcatgcca
     4321 taccgcgaaa ggttttgcgc cattcgatac tagtagcggc cggatggatc tcgatcccgc
     4381 gaaat
//
