LOCUS       sequence-300219-        3986 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 3986)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3986)
  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-m4136463
FEATURES             Location/Qualifiers
     source          1..3986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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             78..100
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             111..128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             447..467
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      840..871
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        872..974
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(975..1043,1044..1835)
                     /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(1193..1212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1836..2027
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      2051..2078
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2090..2677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2170..2189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2699..2728)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(2814..3896)
                     /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 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg caccatcatc
      121 accatcatgg tggttctggt atgtctgatt ctgaagttaa ccaagaagct aaacctgagg
      181 ttaagccaga agtcaaacca gaaactcata ttaacttgaa ggtttctgat ggttcttctg
      241 aaatcttctt caagatcaag aagaccactc cattgcgtag attgatggaa gctttcgcta
      301 aaagacaagg taaagaaatg gattctttga gattcttgta cgacggtatc agaattcaag
      361 ctgatcaaac tcctgaagat ttggatatgg aggataatga catcatcgaa gctcatagag
      421 aacaaattgg tggtggtggt tctggtgaga acctgtactt tcaaagctat agttttgtat
      481 cagaggaaac ggggacactt attgtcaata gcgtgttgct gttcctagct ttcgttgttt
      541 ttctgctcgt gaccttagcg atcctgactg ccttgcgtct gtgtgcatac tgctgcaaca
      601 tcgttaacgt atccctggta aagccgtctt tctacgtgta ttcgcgcgtt aaaaacctca
      661 acagctctag ggttcccgac ctgcttgttt aaagagaata taaaaagcca gattattaat
      721 ccggcttttt tattatttag gttctgttaa gtaactgaac ccaatgtcgt tagtgacgct
      781 tacctcttaa gaggtcactg acctaacagc ttgactccag cgtaactgga ctgcaatcaa
      841 ctcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt tccaactttc
      901 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
      961 ctaaggaagc taaaatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1021 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1081 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1141 agagtttacg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1201 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1261 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatctcacg gatggcatga
     1321 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1381 ttctggcaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1441 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1501 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1561 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1621 gatcacttct gcgctcggcc ctcccggctg gctggtttat tgctgataaa tctggagccg
     1681 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgca
     1741 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1801 ctgagatagg tgcctcactg attaagcatt ggtaagtgac caaacaggaa aaaaccgccc
     1861 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     1921 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     1981 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgatga gggcccaaat
     2041 gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt ttccataggc
     2101 tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg cgaaacccga
     2161 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2221 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2281 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2341 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2401 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2461 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2521 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc tcggaaaaag
     2581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtc tttttgtttg
     2641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga ttttctaccg
     2701 aagaaaggcc cacccgtgaa ggtgagccag tgactttctg ctatggaggt caggtatgat
     2761 ttaaatggtc agtgcggcgc gccaacaaaa aacccctagc cccccgtttt atatcactgc
     2821 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2881 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagactggc
     2941 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     3001 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3061 ctatcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     3121 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3181 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3241 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     3301 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     3361 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcctcatg ggagaaaata
     3421 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3481 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3541 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3601 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3661 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     3721 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3781 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3841 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcatattc
     3901 accaccctga attgactgtc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     3961 cattcgatac tagtagcggc cggatg
//
