LOCUS       sequence-337714-        8371 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein mink variant 
            (S-GSAS-D614G-deltaFV variant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8371)
  AUTHORS   Gobeil SM, Janowska K, McDowell S, Mansouri K, Parks R, Stalls V, 
            Kopp MF, Manne K, Li D, Wiehe K, Saunders KO, Edwards RJ, Korber B, 
            Haynes BF, Henderson R, Acharya P
  TITLE     Effect of natural mutations of SARS-CoV-2 on spike structure, 
            conformation, and antigenicity.
  JOURNAL   Science. 2021 Jun 24. pii: science.abi6226. doi: 
            10.1126/science.abi6226.
  PUBMED    34168071
REFERENCE   2  (bases 1 to 8371)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2021 Jun 24. pii: science.abi6226. doi: 10.1126/science.abi6226."
COMMENT     Sequence Label: p-alpha-H-S-GSAS-D614G-deltaFV
FEATURES             Location/Qualifiers
     source          1..8371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5481..5504
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5508..5531
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5538..5561
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5601..5624
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5726..5745)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5791..5846
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5845..5864)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6207..6223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6207..6223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6220..6242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6231..6247
                     /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(6255..6261,6262..6279,6280..6285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6300..6321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6450..6467)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6621..7209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6701..6720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7380..8171,8172..8240))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8003..8022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8241..8345)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggncgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagcgcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga gttcggtacc gccaccatgt tcgtgttcct ggtgctcctg cctctggtga
     1801 gcagccagtg cgtgaacctg accacccgaa cccagctccc accagcctac accaacagct
     1861 ttacacgggg cgtgtactac cctgacaagg tgttcagatc tagcgtcctg cacagcactc
     1921 aggacctctt cctgccgttc ttcagcaacg tgacatggtt ccacgccatc agcggcacaa
     1981 acggaaccaa gcggtttgat aaccccgtcc tgccattcaa tgatggagtt tacttcgcca
     2041 gtaccgagaa gagtaacatc atccggggct ggatcttcgg caccaccctg gatagcaaaa
     2101 cacagagcct cctgatcgtg aacaatgcca cgaacgtcgt gatcaaggtg tgcgagttcc
     2161 agttttgcaa tgatcctttc ctgggtgtgt actaccacaa gaacaacaag agctggatgg
     2221 aaagcgagtt cagagtctac agcagcgcca acaactgcac attcgagtac gtctctcagc
     2281 cttttctgat ggaccttgag gggaaacaag gcaacttcaa gaacctgaga gaattcgtgt
     2341 tcaagaacat cgacggctac ttcaaaatct actccaagca cacacccatc aacctggtcc
     2401 gggacctccc tcagggcttc agcgccctgg aacccctggt cgacctgccc ataggcatca
     2461 acataacgcg gttccaaacc ctgctggccc tgcatagatc ctacctgact cctggcgaca
     2521 gcagcagcgg atggaccgcc ggagctgcag cctactatgt gggctacctg caacctagaa
     2581 ccttcctgct gaagtacaac gagaacggca caatcacaga cgccgtcgac tgcgccctgg
     2641 accctctctc tgagacaaag tgcaccctga agtccttcac cgtggaaaag ggcatctacc
     2701 agaccagcaa cttccgggtg cagcctacag agagcatcgt gcgatttcca aacattacca
     2761 acctctgccc cttcggcgag gtgtttaacg ccacaagatt tgcctccgtt tacgcctgga
     2821 atagaaagag aatcagcaat tgtgtggccg actactccgt gctgtataac agcgcctctt
     2881 tcagcacctt caagtgctac ggcgtttccc caacaaagct gaatgacctg tgcttcacca
     2941 acgtgtacgc cgactccttc gtaattagag gcgatgaggt gcggcagatc gcaccaggcc
     3001 agaccggtaa gatcgctgac tacaactata agctgcctga tgattttaca ggctgcgtga
     3061 tcgcctggaa ctctaacaac ctggatagca aggtgggcgg caactacaac tacctgttcc
     3121 ggctgtttcg caagtctaac ctgaaacctt tcgagagaga catctccaca gagatctacc
     3181 aggccggttc tacaccttgt aacggggtgg aaggcttcaa ctgttacttc cctctgcaaa
     3241 gctacggctt ccagcctacc aatggagtcg gctaccagcc ataccgggtg gtcgtgctgt
     3301 ccttcgagtt actccacgcc cccgccaccg tctgcggtcc taagaagtcc accaatctgg
     3361 ttaagaacaa atgcgtgaac ttcaacttca acggcctgac cgggaccggc gtgctgaccg
     3421 aaagcaacaa aaagttcctc cccttccagc agttcggccg tgatatcgct gacaccacag
     3481 atgccgtcag agatccacag accctggaaa tcctggatat tacaccctgc tccttcggag
     3541 gagtttctgt gatcaccccc gggaccaata ccagcaacca ggtggctgtg ctgtaccaag
     3601 gtgttaactg caccgaggtt cctgtggcca tccacgccga tcagctgaca cctacttgga
     3661 gagtgtactc cactggctcc aatgtgttcc agaccagggc cggatgtctg atcggcgccg
     3721 agcacgtgaa taacagttac gagtgcgaca tccctatcgg cgccggcatc tgtgccagct
     3781 accagaccca gacaaacagc cctgggtctg cttcctctgt agctagccag agcgtcatcg
     3841 cctacaccat gagcctgggc gcagagaaca gcgtggccta ttccaacaac tctatcgcca
     3901 ttcccaccaa ctttacaatt agcgtcacaa cagagatcct gcccgtgagc atgaccaaga
     3961 ccagcgtgga ctgtacaatg tacatctgtg gcgacagcac tgaatgcagc aacctgctgc
     4021 tgcaatacgg ctccttttgc acccaactga accgggcgct gaccggaatc gccgtggaac
     4081 aggacaaaaa tacccaggag gtgttcgccc aagtgaagca gatctacaag accccaccta
     4141 tcaaggactt cggcggcttt aactttagcc agattctccc tgatccttct aagcctagca
     4201 agcggagctt tatcgaggat ctgctgttca acaaggtcac cctggccgat gccggcttta
     4261 tcaaacagta tggcgattgc ctgggcgaca tagccgccag agatctgatc tgcgcccaga
     4321 aattcaacgg cctgacagtt ctcccacctc tgctgaccga cgagatgatc gctcagtaca
     4381 cctctgccct gctggctggc accatcacat ctgggtggac atttggcgcc ggcgccgccc
     4441 tgcagatccc ctttgccatg cagatggcct atagattcaa cggaatcggc gtgacccaga
     4501 acgtgctgta tgaaaaccag aagctgatcg ctaaccagtt caattctgcc atcggcaaga
     4561 tccaggactc cctctcctct accgccagcg ccctgggcaa actgcaggac gtggtgaatc
     4621 agaacgccca agccctgaac accctggtga agcagctcag cagcaatttt ggcgccatca
     4681 gctctgtgct gaacgatatc ctgtctagac tggacaaggt ggaagccgaa gtccagatcg
     4741 atagactgat cacaggcaga ctgcagtccc tgcaaaccta cgtgacccaa cagctgatca
     4801 gggccgctga aataagagcc agcgccaatc tcgccgctac caagatgtcc gagtgtgtgc
     4861 tgggacagtc taaacgcgtt gacttctgcg gcaaaggcta tcacctgatg agcttccccc
     4921 agagcgcgcc gcacggcgtg gtgttcctgc atgtgacata cgtgcctgcc caagagaaga
     4981 atttcacaac cgcccctgcc atctgccacg acggcaaggc ccacttccca agagagggcg
     5041 ttttcgtttc caatggcaca cactggttcg tgacacaaag aaacttctac gaaccccaga
     5101 ttatcaccac cgacaacacc ttcgtgagtg gcaattgtga cgtggtcatc ggaatcgtga
     5161 acaacacagt gtacgaccct ctgcaacctg agctggactc ttttaaggaa gagctggaca
     5221 agtactttaa aaaccacacc agccccgatg tggacctggg cgacatcagt ggcattaacg
     5281 ccagcgtggt gaacatccaa aaggaaatcg acagactgaa cgaggtggcc aagaacctga
     5341 acgagtccct gatcgacctg caggagctcg gcaaatacga gcagggatcc ggatacatcc
     5401 ccgaggcccc cagagatggc caggcctacg tgcggaagga cggcgagtgg gtactgctga
     5461 gcacattcct gggcagatcc ctggaggtgc tgttccaggg cccaggccat caccaccatc
     5521 accaccatca tagcgcctgg tcccaccccc agttcgagaa gggcggcggt agtggagggg
     5581 gcggatctgg cggctcagct tggagccacc cccagttcga aaagtgataa tgactcgagc
     5641 gataattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     5701 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     5761 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     5821 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     5881 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     5941 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     6001 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     6061 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     6121 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     6181 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6241 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6301 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6361 ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     6421 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6481 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6541 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6601 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6661 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6721 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6781 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6841 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6901 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6961 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7021 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7081 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7141 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7201 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7261 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7321 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7381 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7441 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7501 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7561 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7621 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7681 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7741 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7801 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7861 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7921 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7981 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8041 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8101 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8161 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8221 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8281 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8341 ccgcgcacat ttccccgaaa agtgccacct g
//
