LOCUS       Exported                8996 bp ds-DNA     circular SYN 08-APR-2022
DEFINITION  Expresses virus-like particles based upon the original SARS-Cov-2.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Spike-VLP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8996)
  AUTHORS   Andreas Sturmer
  TITLE     Plasmids expressing the original SARS-Coronavirus 2 Spike Protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 8, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8996
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    4879..5103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      5558..5567
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    6543..6598
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     polyA_signal    6616..6737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6653..6672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6707..6726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    6753..6824
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     protein_bind    6825..6858
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(6906..6922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6906..6922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6919..6941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6930..6946
                     /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(6954..6984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6999..7020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7137..7154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7308..7896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7388..7407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8074..8883)
                     /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     8797..8816
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8875..8894
                     /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 atctagaaaa caagctagct gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4921 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4981 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5041 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     5101 tggaattcca ttgacccaat gtacttgggc aaaacccagt ggaaagtccc atttgactca
     5161 gtgtgcttgc cagtaatggg aaaaacccat tggcttacct cccattgacc caatgtactt
     5221 gggtaaaggc cattgagtca ccacccctat gctgggaaat ggtgaacgcc ccctatgtgg
     5281 aaagtcccta tgggccgcca ttagagtgca tgaccgtgct gattcatatg ccatatgagt
     5341 gtattagggg gctttccgct tgggaaattg ggtaaaaagt ccccgtatta ctcacatagg
     5401 gggcgtttgg ctttgcaaat taggggattt tagtgcattt ggcattaaaa actattggtt
     5461 ctagtcataa aacgggcgga gttaaccata taaaagctgt cccccatgcc attcgagccc
     5521 agagcaggac ggacctaagc tcgatacgac cctatcagcc accatggcag attccaacgg
     5581 tactattacc gttgaagagc ttaagaagct ccttgaacaa tggaacctag taataggttt
     5641 cctattcctt acatggattt gtcttctaca atttgcctat gccaacagga ataggttctt
     5701 gtatataatt aagttaattt tcctctggct gttatggcca gtaactttag cttgttttgt
     5761 gcttgctgct gtttacagaa taaattggat cactggtgga attgctatcg caatggcttg
     5821 tcttgtaggc ttgatgtggc tcagctactt cattgcttct ttcagactgt ttgcgcgtac
     5881 gcgttccatg tggtcattca atccagaaac taacattctt ctcaacgtgc cactgcatgg
     5941 cactattctg accagaccgc tgctagaaag tgaactcgta atcggagctg tgatccttcg
     6001 tggacatctt cgtattgctg gacaccatct aggacgctgt gacatcaagg acctgcctaa
     6061 agaaatcact gttgctacat cacgaacgct ttcttattac aaattgggag cttcgcagcg
     6121 tgtagcaggt gactcaggtt ttgctgcata cagtcgctac aggattggca actataaatt
     6181 aaacacagac cattccagta gcagtgacaa tattgctttg cttgtacagt aaggtacctc
     6241 aattaaacat atttataatt tcttcttcca gaacacctct gaaagcttgg aacaaactaa
     6301 gccaccatgt actcattcgt ttcggaagag acaggtacgt taatagttaa tagcgtactt
     6361 ctttttcttg ctttcgtggt attcttgcta gttacactag ccatccttac tgcgcttcga
     6421 ttgtgtgcgt actgctgcaa tattgttaac gtgagtcttg taaaaccttc tttttacgtt
     6481 tactctcgtg ttaagaatct caattcttcc agagttcctg atcttctggt ctaaatctaa
     6541 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcaca
     6601 attgaagagc tcctgaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6661 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6721 ctcatcaatg tatcttatag tagagctggt accattcctt taatggtctt ttctggacac
     6781 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaataact tcgtatagca
     6841 tacattatac gaagttatga tatcgtcgac tgcagaggcc tgcatgcaag cttggtgtaa
     6901 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7021 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7081 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     7141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7741 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7861 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7981 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aagcccaatc
     8041 tgaataatgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     8101 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     8161 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     8221 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     8281 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     8341 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     8401 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     8461 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     8521 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     8581 tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     8641 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     8701 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     8761 caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     8821 taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct
     8881 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     8941 atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga gctgca
//
