LOCUS       sequence-313545-        5558 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 5558)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5558)
  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-m4046925
FEATURES             Location/Qualifiers
     source          1..5558
                     /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..3952
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      4073..4661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4562..4581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4767..5426
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4833..4852)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5497..5531
                     /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 ttcaaagctg gagccatccg cagtttgaaa aataaccggc
     3961 ttatcggtca gtttcacctg atttacgtaa aaacccgctt cggcgggttt ttgcttttgg
     4021 aggggcagaa agatgaatga ctgtccacga cgctataccc aaaagaaaaa gcttgagatc
     4081 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4141 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4201 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4261 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4321 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4381 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4441 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4501 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4561 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4621 gatttttgtg atgctcgtca ggggggcgga gcctatggaa attgatcggg cacgtaagag
     4681 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     4741 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     4801 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     4861 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     4921 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     4981 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     5041 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     5101 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     5161 cctatttccc taaagggttt attgagaata tgtttttcgt ttcagccaat ccctgggtga
     5221 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     5281 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     5341 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     5401 gcgatgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact cctgttgata
     5461 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     5521 gcgtttttta ttggtcatcg atctcgatcc cgcgaaat
//
