LOCUS       sequence-298346-        5552 bp    DNA     circular SYN 14-APR-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 5552)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5552)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046828
FEATURES             Location/Qualifiers
     source          1..5552
                     /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             3927..3947
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3948..3965
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      4086..4674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4575..4594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4780..5439
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4846..4865)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5510..5544
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg tttgtcttcc
      121 tggtactcct tccgttggtg tccagtcaat gcgttaacct gaccacgaga actcagctgc
      181 cacccgcgta taccaattct ttcactaggg gtgtttacta cccggataaa gtatttcggt
      241 cgagcgtttt acattctaca caggacctgt tcctgccttt cttctcaaac gtgacttggt
      301 ttcacgccat tcacgttagc ggcaccaacg ggaccaagcg cttcgacaac ccggtcctgc
      361 cgttcaatga tggtgtgtac tttgcatcta ctgagaaaag caacatcata cgcggctgga
      421 tcttcggaac gactctggac tccaaaaccc agtctctcct gatcgtgaac aatgctacta
      481 acgtagttat taaggtatgt gaatttcaat tctgcaacga tccgttcttg ggcgtatatt
      541 accataaaaa caacaaatct tggatggaat ccgaatttcg tgtttattcc agcgctaata
      601 attgcacttt cgagtacgtt agccagcctt tcctgatgga cctggaaggc aaacagggta
      661 acttcaaaaa cctgcgtgaa tttgttttca agaacatcga tggttacttt aaaatctatt
      721 ctaagcacac cccaattaac ctggttcgtg acctgccgca gggtttctcg gctttggagc
      781 cgctggtaga tcttccgatc gggattaaca tcacccgttt tcaaaccctg ctggcgctgc
      841 accgttcata cctgacaccg ggtgacagtt ctagcggctg gacagcaggc gctgcggcct
      901 actatgtggg ttacctgcaa ccgcgcacct tcctactcaa atacaacgaa aatggcacga
      961 tcaccgacgc cgtcgattgt gcacttgacc ctctgagcga aactaaatgc actctgaaat
     1021 cctttacggt cgaaaaaggt atctatcaga cctccaactt ccgcgttcag cctactgaaa
     1081 gtatcgtacg tttcccgaac attactaacc tgtgtccctt tggagaggtt tttaacgcta
     1141 ccaggttcgc gtctgtgtat gcttggaacc gtaagcgaat ctcaaactgc gtagcggatt
     1201 acagcgtcct gtataactct gcaagctttt cgactttcaa atgctacggc gtgtctccaa
     1261 ccaagttgaa cgatctatgt ttcaccaacg tctacgctga ttctttcgtt atccgtggtg
     1321 acgaggttcg tcagattgca ccgggccaga caggtaaaat cgcggattac aattataaat
     1381 taccggacga cttcacgggc tgcgtaatag cttggaactc taacaatctg gatagcaaag
     1441 ttggcggtaa ctacaactac ttatatcgac tgttccgcaa aagcaacctg aagcccttcg
     1501 aacgcgacat ctccactgaa atttaccagg ccggttcgac cccgtgcaac ggtgtagaag
     1561 gttttaattg ctacttcccg ttgcagtcct atggctttca gccgaccaac ggcgttggct
     1621 atcagcctta ccgtgttgtg gtgctctcct ttgaactgct gcatgcccct gcaaccgtat
     1681 gtggcccgaa aaaatccact aacttagtga aaaacaaatg cgtaaacttt aatttcaatg
     1741 gtctgacagg gacgggcgtc ctgactgaat ctaacaaaaa gttcctgcca ttccaacagt
     1801 ttggtcgcga tattgctgac actaccgatg cagttcgtga cccgcagacg ctggaaatcc
     1861 tggatatcac cccgtgttct ttcggtggcg ttagcgtgat cactccgggt acaaacacct
     1921 ccaaccaggt ggctgtcctg taccaggacg ttaactgcac ggaagtaccg gtggcaattc
     1981 acgctgatca gctgactccg acgtggcgtg tctactctac cggttcgaat gttttccaaa
     2041 cgcgcgctgg ctgcctgatt ggtgctgaac atgtaaacaa ttcttacgag tgcgacatcc
     2101 ccataggagc gggtatctgt gcgagctacc agactcagac caacagccct cgtcgcgccc
     2161 gttccgtggc aagccagtca atcatcgcgt atacgatgag cctgggtgcc gagaactcgg
     2221 ttgcatacag taacaatagc attgctatcc caactaactt cacaatttca gttaccacag
     2281 aaatccttcc ggtatctatg actaaaactt ccgtggattg caccatgtac atttgcggcg
     2341 acagcactga gtgttccaat cttttactgc aatatggtag tttttgcacg cagctgaacc
     2401 gtgccctgac cggaatcgcg gttgaacagg ataaaaacac ccaggaagtt ttcgctcaag
     2461 taaaacagat ttataagacc ccgcccatca aagacttcgg cggttttaat ttcagccaaa
     2521 tcctccccga cccgtctaag ccaagcaaac gctcattcat cgaagacctg ctgtttaaca
     2581 aagtgactct ggcggacgct ggcttcatca agcaatacgg agactgtcta ggggatattg
     2641 cagctcgtga cctcatttgc gcgcagaagt tcaacggctt aactgttctg ccgccgctgc
     2701 tgaccgatga gatgatcgct cagtacacat ccgctctgct ggcaggtacc atcacctctg
     2761 ggtggacgtt tggcgcgggt gccgctctcc agattccatt cgccatgcaa atggcttatc
     2821 gctttaatgg aatcggcgta actcagaacg ttttgtacga aaaccagaaa ctgatagcta
     2881 accaattcaa ttctgcgatt ggcaaaatcc aggactctct gagctccact gcaagcgctc
     2941 tcggcaaact gcaagacgtt gtcaaccaga acgcgcaggc actgaacacc ctggtaaaac
     3001 agctgtcaag caacttcggt gctatcagtt cagttttaaa tgatatcctg tcgcggctgg
     3061 ataaagttga agccgaagtt caaatcgacc gtctgattac cggtcgtctg caatctctgc
     3121 aaacttatgt aacccagcag ctgatcagag ctgccgagat tcgcgctagc gcaaacttgg
     3181 cggcgacaaa aatgtcagaa tgcgtacttg gccagtctaa acgtgtggac ttctgcggta
     3241 aaggttacca cctgatgtcg tttccgcagt cggcccctca tggtgtagtt ttcctacacg
     3301 tgacttacgt cccggcccag gaaaagaact ttaccactgc tccagccatc tgccacgatg
     3361 gtaaagctca tttcccgcgt gaaggcgtct tcgtttccaa cggcactcac tggttcgtga
     3421 cgcagcgtaa cttctacgag ccgcagatca ttactactga taacaccttt gtttcgggta
     3481 actgcgatgt cgtgatcggc atcgttaata acactgttta cgacccgctg caaccggaac
     3541 tggattcctt taaagaagaa ctggataaat attttaaaaa ccacacctct ccagatgtag
     3601 acctgggaga cattagtggc ataaatgcgt ccgttgttaa cattcagaaa gaaatcgacc
     3661 gcctgaacga ggtcgcaaag aatctgaacg agtctctgat tgacctgcaa gagctgggta
     3721 agtatgagca gtacattaaa tggccttggt acatctggct gggcttcatc gcaggtttga
     3781 tcgccatcgt catggtgact attatgctct gctgtatgac gtcttgctgc tcctgcctga
     3841 aaggttgttg ctcttgcggg tcgtgttgca agtttgacga agatgacagt gaaccggttc
     3901 tgaaaggcgt taaacttcat tacaccgaga acctgtactt tcaaagccac catcatcacc
     3961 atcattaacc ggcttatcgg tcagtttcac ctgatttacg taaaaacccg cttcggcggg
     4021 tttttgcttt tggaggggca gaaagatgaa tgactgtcca cgacgctata cccaaaagaa
     4081 aaagcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4141 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4201 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4261 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4321 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4381 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4441 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4501 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4561 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4621 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaattgatc
     4681 gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     4741 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     4801 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     4861 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     4921 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     4981 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     5041 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     5101 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     5161 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtttcagcc
     5221 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     5281 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     5341 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     5401 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg
     5461 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     5521 cggttgccgc cgggcgtttt ttattggtca tc
//
