LOCUS       sequence-311539-        5561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 surface glycoprotein 
            (Spike protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5561)
  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-m4046970
FEATURES             Location/Qualifiers
     source          1..5561
                     /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             3908..3928
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3929..3955
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      4076..4664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4565..4584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4770..5429
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4836..4855)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5500..5534
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat gtttgtcttc ctggtactcc ttccgttggt
      121 gtccagtcaa tgcgttaacc tgaccacgag aactcagctg ccacccgcgt ataccaattc
      181 tttcactagg ggtgtttact acccggataa agtatttcgg tcgagcgttt tacattctac
      241 acaggacctg ttcctgcctt tcttctcaaa cgtgacttgg tttcacgcca ttcacgttag
      301 cggcaccaac gggaccaagc gcttcgacaa cccggtcctg ccgttcaatg atggtgtgta
      361 ctttgcatct actgagaaaa gcaacatcat acgcggctgg atcttcggaa cgactctgga
      421 ctccaaaacc cagtctctcc tgatcgtgaa caatgctact aacgtagtta ttaaggtatg
      481 tgaatttcaa ttctgcaacg atccgttctt gggcgtatat taccataaaa acaacaaatc
      541 ttggatggaa tccgaatttc gtgtttattc cagcgctaat aattgcactt tcgagtacgt
      601 tagccagcct ttcctgatgg acctggaagg caaacagggt aacttcaaaa acctgcgtga
      661 atttgttttc aagaacatcg atggttactt taaaatctat tctaagcaca ccccaattaa
      721 cctggttcgt gacctgccgc agggtttctc ggctttggag ccgctggtag atcttccgat
      781 cgggattaac atcacccgtt ttcaaaccct gctggcgctg caccgttcat acctgacacc
      841 gggtgacagt tctagcggct ggacagcagg cgctgcggcc tactatgtgg gttacctgca
      901 accgcgcacc ttcctactca aatacaacga aaatggcacg atcaccgacg ccgtcgattg
      961 tgcacttgac cctctgagcg aaactaaatg cactctgaaa tcctttacgg tcgaaaaagg
     1021 tatctatcag acctccaact tccgcgttca gcctactgaa agtatcgtac gtttcccgaa
     1081 cattactaac ctgtgtccct ttggagaggt ttttaacgct accaggttcg cgtctgtgta
     1141 tgcttggaac cgtaagcgaa tctcaaactg cgtagcggat tacagcgtcc tgtataactc
     1201 tgcaagcttt tcgactttca aatgctacgg cgtgtctcca accaagttga acgatctatg
     1261 tttcaccaac gtctacgctg attctttcgt tatccgtggt gacgaggttc gtcagattgc
     1321 accgggccag acaggtaaaa tcgcggatta caattataaa ttaccggacg acttcacggg
     1381 ctgcgtaata gcttggaact ctaacaatct ggatagcaaa gttggcggta actacaacta
     1441 cttatatcga ctgttccgca aaagcaacct gaagcccttc gaacgcgaca tctccactga
     1501 aatttaccag gccggttcga ccccgtgcaa cggtgtagaa ggttttaatt gctacttccc
     1561 gttgcagtcc tatggctttc agccgaccaa cggcgttggc tatcagcctt accgtgttgt
     1621 ggtgctctcc tttgaactgc tgcatgcccc tgcaaccgta tgtggcccga aaaaatccac
     1681 taacttagtg aaaaacaaat gcgtaaactt taatttcaat ggtctgacag ggacgggcgt
     1741 cctgactgaa tctaacaaaa agttcctgcc attccaacag tttggtcgcg atattgctga
     1801 cactaccgat gcagttcgtg acccgcagac gctggaaatc ctggatatca ccccgtgttc
     1861 tttcggtggc gttagcgtga tcactccggg tacaaacacc tccaaccagg tggctgtcct
     1921 gtaccaggac gttaactgca cggaagtacc ggtggcaatt cacgctgatc agctgactcc
     1981 gacgtggcgt gtctactcta ccggttcgaa tgttttccaa acgcgcgctg gctgcctgat
     2041 tggtgctgaa catgtaaaca attcttacga gtgcgacatc cccataggag cgggtatctg
     2101 tgcgagctac cagactcaga ccaacagccc tcgtcgcgcc cgttccgtgg caagccagtc
     2161 aatcatcgcg tatacgatga gcctgggtgc cgagaactcg gttgcataca gtaacaatag
     2221 cattgctatc ccaactaact tcacaatttc agttaccaca gaaatccttc cggtatctat
     2281 gactaaaact tccgtggatt gcaccatgta catttgcggc gacagcactg agtgttccaa
     2341 tcttttactg caatatggta gtttttgcac gcagctgaac cgtgccctga ccggaatcgc
     2401 ggttgaacag gataaaaaca cccaggaagt tttcgctcaa gtaaaacaga tttataagac
     2461 cccgcccatc aaagacttcg gcggttttaa tttcagccaa atcctccccg acccgtctaa
     2521 gccaagcaaa cgctcattca tcgaagacct gctgtttaac aaagtgactc tggcggacgc
     2581 tggcttcatc aagcaatacg gagactgtct aggggatatt gcagctcgtg acctcatttg
     2641 cgcgcagaag ttcaacggct taactgttct gccgccgctg ctgaccgatg agatgatcgc
     2701 tcagtacaca tccgctctgc tggcaggtac catcacctct gggtggacgt ttggcgcggg
     2761 tgccgctctc cagattccat tcgccatgca aatggcttat cgctttaatg gaatcggcgt
     2821 aactcagaac gttttgtacg aaaaccagaa actgatagct aaccaattca attctgcgat
     2881 tggcaaaatc caggactctc tgagctccac tgcaagcgct ctcggcaaac tgcaagacgt
     2941 tgtcaaccag aacgcgcagg cactgaacac cctggtaaaa cagctgtcaa gcaacttcgg
     3001 tgctatcagt tcagttttaa atgatatcct gtcgcggctg gataaagttg aagccgaagt
     3061 tcaaatcgac cgtctgatta ccggtcgtct gcaatctctg caaacttatg taacccagca
     3121 gctgatcaga gctgccgaga ttcgcgctag cgcaaacttg gcggcgacaa aaatgtcaga
     3181 atgcgtactt ggccagtcta aacgtgtgga cttctgcggt aaaggttacc acctgatgtc
     3241 gtttccgcag tcggcccctc atggtgtagt tttcctacac gtgacttacg tcccggccca
     3301 ggaaaagaac tttaccactg ctccagccat ctgccacgat ggtaaagctc atttcccgcg
     3361 tgaaggcgtc ttcgtttcca acggcactca ctggttcgtg acgcagcgta acttctacga
     3421 gccgcagatc attactactg ataacacctt tgtttcgggt aactgcgatg tcgtgatcgg
     3481 catcgttaat aacactgttt acgacccgct gcaaccggaa ctggattcct ttaaagaaga
     3541 actggataaa tattttaaaa accacacctc tccagatgta gacctgggag acattagtgg
     3601 cataaatgcg tccgttgtta acattcagaa agaaatcgac cgcctgaacg aggtcgcaaa
     3661 gaatctgaac gagtctctga ttgacctgca agagctgggt aagtatgagc agtacattaa
     3721 atggccttgg tacatctggc tgggcttcat cgcaggtttg atcgccatcg tcatggtgac
     3781 tattatgctc tgctgtatga cgtcttgctg ctcctgcctg aaaggttgtt gctcttgcgg
     3841 gtcgtgttgc aagtttgacg aagatgacag tgaaccggtt ctgaaaggcg ttaaacttca
     3901 ttacaccgag aacctgtact ttcaaagcta cccgtatgac gttcctgatt atgcctaacc
     3961 ggcttatcgg tcagtttcac ctgatttacg taaaaacccg cttcggcggg tttttgcttt
     4021 tggaggggca gaaagatgaa tgactgtcca cgacgctata cccaaaagaa aaagcttgag
     4081 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4141 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4201 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4261 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4321 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4381 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4441 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4501 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4561 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4621 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaattgatc gggcacgtaa
     4681 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     4741 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     4801 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     4861 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     4921 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     4981 cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     5041 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     5101 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     5161 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtttcagcc aatccctggg
     5221 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     5281 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     5341 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     5401 actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg
     5461 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     5521 cgggcgtttt ttattggtca tcgatctcga tcccgcgaaa t
//
