LOCUS       sequence-313668-        4790 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 4790)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4790)
  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-m4136460
FEATURES             Location/Qualifiers
     source          1..4790
                     /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..1219
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1076..1085
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1193..1216
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1232..1252
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1625..1656
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1657..1759
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(1760..1828,1829..2620)
                     /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(1978..1997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2621..2812
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      2836..2863
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2875..3462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2955..2974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3484..3513)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(3599..4681)
                     /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 taaaatcgaa gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct
      181 cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc
      241 ggataaactg gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat
      301 cttctgggca cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac
      361 cccggacaaa gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa
      421 cggcaagctg attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga
      481 tctgctgccg aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa
      541 agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct
      601 gattgctgct gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga
      661 cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa
      721 aaacaaacac atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg
      781 cgaaacagcg atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt
      841 gaattatggt gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg
      901 cgtgctgagc gcaggtatta acgccgccag tccgaacaaa gagctggcga aagagttcct
      961 cgaaaactat ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg
     1021 tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac
     1081 catggaaaac gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg
     1141 gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc
     1201 cctgaaagac gcgcagactg gtggttctgg tgagaacctg tactttcaaa gctatagttt
     1261 tgtatcagag gaaacgggga cacttattgt caatagcgtg ttgctgttcc tagctttcgt
     1321 tgtttttctg ctcgtgacct tagcgatcct gactgccttg cgtctgtgtg catactgctg
     1381 caacatcgtt aacgtatccc tggtaaagcc gtctttctac gtgtattcgc gcgttaaaaa
     1441 cctcaacagc tctagggttc ccgacctgct tgtttaaaga gaatataaaa agccagatta
     1501 ttaatccggc ttttttatta tttaggttct gttaagtaac tgaacccaat gtcgttagtg
     1561 acgcttacct cttaagaggt cactgaccta acagcttgac tccagcgtaa ctggactgca
     1621 atcaactcaa aagcctccga ccggaggctt ttgacttgat cggcacgtaa gaggttccaa
     1681 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     1741 aggagctaag gaagctaaaa tgagtattca acatttccgt gtcgccctta ttcccttttt
     1801 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1861 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1921 ccttgagagt ttacgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1981 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2041 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc tcacggatgg
     2101 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2161 cttacttctg gcaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2221 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2281 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2341 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2401 tgcaggatca cttctgcgct cggccctccc ggctggctgg tttattgctg ataaatctgg
     2461 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2521 ccgcatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2581 gatcgctgag ataggtgcct cactgattaa gcattggtaa gtgaccaaac aggaaaaaac
     2641 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     2701 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     2761 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gatgagggcc
     2821 caaatgtaat cacctggctc accttcgggt gggcctttct gcgttgctgg cgtttttcca
     2881 taggctccgc ccccctgacg agcatcacaa aaatcgatgc tcaagtcaga ggtggcgaaa
     2941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttacctcgga
     3361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggtcttttt
     3421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgattttc
     3481 taccgaagaa aggcccaccc gtgaaggtga gccagtgact ttctgctatg gaggtcaggt
     3541 atgatttaaa tggtcagtgc ggcgcgccaa caaaaaaccc ctagcccccc gttttatatc
     3601 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3661 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     3721 ctggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     3781 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     3841 atgagctatc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     3901 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     3961 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4021 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     4081 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     4141 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtcc tcatgggaga
     4201 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     4261 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     4321 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     4381 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     4441 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     4501 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     4561 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     4621 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     4681 tattcaccac cctgaattga ctgtcttccg ggcgctatca tgccataccg cgaaaggttt
     4741 tgcgccattc gatactagta gcggccggat ggatctcgat cccgcgaaat
//
