LOCUS       sequence-358849-        7286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the original SARS-Cov-2 Spike Protein under a CMV 
            Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7286)
  AUTHORS   Andreas Sturmer
  TITLE     Plasmids expressing the original SARS-Coronavirus 2 Spike Protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7286)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCMV-Spike
FEATURES             Location/Qualifiers
     source          1..7286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    407..478
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     CDS             complement(469..486)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     enhancer        487..790
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        791..994
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     945..965
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    4906..5027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4943..4962)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4997..5016
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    5043..5114
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     protein_bind    complement(5115..5148)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5196..5212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5196..5212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5209..5231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5220..5236
                     /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)."
     promoter        complement(join(5244..5250,5251..5268,5269..5274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5427..5444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5598..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5678..5697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6364..7173)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7087..7106
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7165..7184
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgatatcatc ttggaccatt
      421 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcgg
      481 taccagcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      541 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      601 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      661 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      721 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      781 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      841 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      901 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      961 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     1021 gcgctaccgg tcgccaccat gtttgttttt cttgttttat tgccactagt ctctagtcag
     1081 tgtgttaatc ttacaaccag aactcaatta cctcctgcat acactaattc tttcacacgt
     1141 ggtgtttatt accctgacaa agttttcaga tcctcagttt tacattcaac tcaggacttg
     1201 ttcttacctt tcttttccaa tgttacttgg ttccatgcta tacatgtctc tgggaccaat
     1261 ggtactaaga ggtttgataa ccctgtccta ccatttaatg atggtgttta ttttgcttcc
     1321 actgagaagt ctaacataat aagaggctgg atttttggta ctactttaga ttcgaagacc
     1381 cagtccctac ttattgttaa taacgctact aatgttgtta ttaaagtctg tgaatttcaa
     1441 ttttgtaatg atccattttt gggtgtttat taccacaaaa acaacaaaag ttggatggaa
     1501 agtgagttca gagtttattc tagtgcgaat aattgcactt ttgaatatgt ctctcagcct
     1561 tttcttatgg accttgaagg aaaacagggt aatttcaaaa atcttaggga atttgtgttt
     1621 aagaatattg atggttattt taaaatatat tctaagcaca cgcctattaa tttagtgcgt
     1681 gatctccctc agggtttttc ggctttagaa ccattggtag atttgccaat aggtattaac
     1741 atcactaggt ttcaaacttt acttgcttta catagaagtt atttgactcc tggtgattct
     1801 tcttcaggtt ggacagctgg tgctgcagct tattatgtgg gttatcttca acctaggact
     1861 tttctattaa aatataatga aaatggaacc attacagatg ctgtagactg tgcacttgac
     1921 cctctctcag aaacaaagtg tacgttgaaa tccttcactg tagaaaaagg aatctatcaa
     1981 acttctaact ttagagtcca accaacagaa tctattgtta gatttcctaa tattacaaac
     2041 ttgtgccctt ttggtgaagt ttttaacgcc accagatttg catctgttta tgcttggaac
     2101 aggaagagaa tcagcaactg tgttgctgat tattctgtcc tatataattc cgcatcattt
     2161 tccactttta agtgttatgg agtgtctcct actaaattaa atgatctctg ctttactaat
     2221 gtctatgcag attcatttgt aattagaggt gatgaagtca gacaaatcgc tccagggcaa
     2281 actggaaaga ttgctgatta taattataaa ttaccagatg attttacagg ctgcgttata
     2341 gcttggaact ctaacaatct tgattctaag gttggtggta attataatta cctgtataga
     2401 ttgtttagga agtctaatct caaacctttt gagagagata tttcaactga aatctatcag
     2461 gccggtagca caccttgtaa tggtgttgaa ggttttaatt gttactttcc tttacaatca
     2521 tatggtttcc aacccactaa tggtgttggt taccaaccat acagagtagt agtactttct
     2581 tttgaacttc tacatgcacc agcaactgtt tgtggaccta aaaagtctac taatttggtt
     2641 aaaaacaaat gtgtcaattt caacttcaat ggtttaacag gcacaggtgt tcttactgag
     2701 tctaacaaaa agtttctgcc tttccaacaa tttggcagag acattgctga cactactgat
     2761 gctgtccgtg atccacagac acttgagatt cttgacatta caccatgttc ttttggtggt
     2821 gtcagtgtta taacaccagg aacaaatact tctaaccagg ttgctgttct ttatcaggat
     2881 gttaactgca cagaagtccc tgttgctatt catgcagatc aacttactcc tacttggcgt
     2941 gtttattcta caggttctaa tgtttttcaa acacgtgcag gctgtttaat aggggctgaa
     3001 catgtcaaca actcatatga gtgtgacata cccattggtg caggtatatg cgctagttat
     3061 cagactcaga ctaattctcc tcggcgggca cgtagtgtag ctagtcaatc catcattgcc
     3121 tacactatgt cacttggtgc agaaaattca gttgcttact ctaataactc tattgccata
     3181 cccacaaatt ttactattag tgttaccaca gaaattctac cagtgtctat gaccaagaca
     3241 tcagtagatt gtacaatgta catttgtggt gattcaactg aatgcagcaa tcttttgttg
     3301 caatatggca gtttttgtac acaattaaac cgtgctttaa ctggaatagc tgttgaacaa
     3361 gacaaaaaca cccaagaagt ttttgcacaa gtcaaacaaa tttacaaaac accaccaatt
     3421 aaagattttg gtggttttaa tttttcacaa atattaccag atccatcaaa accaagcaag
     3481 aggtcattta ttgaagatct acttttcaac aaagtgacac ttgcagatgc tggcttcatc
     3541 aaacaatatg gtgattgcct tggtgatatt gctgctagag atctcatttg tgcacaaaag
     3601 tttaacggcc ttactgtttt gccacctttg ctcacagatg aaatgattgc tcaatacact
     3661 tctgcactgt tagcgggtac aatcacttct ggttggacct ttggtgcagg agctgcatta
     3721 caaataccat ttgctatgca aatggcttat aggtttaatg gtattggagt tacacagaat
     3781 gttctctatg agaaccaaaa attgattgcc aaccaattta atagtgctat tggcaaaatt
     3841 caagactcac tttcttccac agcaagtgca cttggaaaac ttcaagatgt ggtcaaccaa
     3901 aatgcacagg ctttaaacac gcttgttaaa caacttagct ccaattttgg tgcaatttca
     3961 agtgttttaa atgacatcct ttcacgtctt gacaaagttg aggctgaagt gcaaattgat
     4021 aggttgatca caggcagact tcaaagtttg cagacatatg tgactcaaca attaattaga
     4081 gctgcagaaa tcagagcttc tgctaatctt gctgctacta aaatgtcaga gtgtgtactt
     4141 ggacaatcaa aaagagttga tttttgtgga aagggctatc atcttatgtc cttccctcag
     4201 tcagcacctc atggtgtagt cttcttgcat gtgacttatg tccctgcaca agaaaagaac
     4261 ttcacaactg ctcctgccat ttgtcatgat ggaaaagcac actttcctcg tgaaggtgtc
     4321 tttgtttcaa atggcacaca ctggtttgta acacaaagga atttttatga accacaaatc
     4381 attactacag acaacacatt tgtgtctggt aactgtgatg ttgtaatagg aattgtcaac
     4441 aacacagttt atgatccttt gcaacctgaa ttagactcat tcaaggagga gttagataaa
     4501 tattttaaga atcatacatc accagatgtt gatttaggtg acatctctgg cattaatgct
     4561 tcagttgtaa acattcaaaa agaaattgac cgcctcaatg aggttgccaa gaatttaaat
     4621 gaatctctca tcgatctcca agaacttgga aagtatgagc agtatataaa atggccatgg
     4681 tacatttggc taggttttat agctggcttg attgccatag taatggtgac aattatgctt
     4741 tgctgtatga ccagttgctg tagttgtctc aagggctgtt gttcttgtgg ttcctgctgc
     4801 aaatttgatg aagacgactc tgagccagtg ctcaaaggag tcaaattaca ttacacataa
     4861 atctagaggg ccctattcta tagtgtcacc taactagagc tcctgaactt gtttattgca
     4921 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4981 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatag tagagctggt
     5041 accattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     5101 ttcttccctc cctaataact tcgtatagca tacattatac gaagttatga tatcgtcgac
     5161 tgcagaggcc tgcatgcaag cttggtgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5221 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5281 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5401 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6001 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gttttaaatc aagcccaatc tgaataatgt tacaaccaat taaccaattc
     6361 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     6421 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     6481 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     6541 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     6601 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     6661 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     6721 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     6781 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     6841 atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga gtaaccatgc
     6901 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     6961 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     7021 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc
     7081 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     7141 cggcctcgac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     7201 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     7261 attttgagac acgggccaga gctgca
//
