LOCUS       40924_48748        8376 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression vector for expression of the SARS-CoV-2 spike 
            HexaPro variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8376)
  AUTHORS   Hsieh CL, Goldsmith JA, Schaub JM, DiVenere AM, Kuo HC, Javanmardi 
            K, Le KC, Wrapp D, Lee AG, Liu Y, Chou CW, Byrne PO, Hjorth CK, 
            Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D, 
            Lavinder JJ, Ippolito GC, Maynard JA, Finkelstein IJ, McLellan JS
  TITLE     Structure-based design of prefusion-stabilized SARS-CoV-2 spikes.
  JOURNAL   Science. 2020 Sep 18;369(6510):1501-1505. doi: 
            10.1126/science.abd0826. Epub 2020 Jul 23.
  PUBMED    32703906
REFERENCE   2  (bases 1 to 8376)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8376)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.abd0826"; journalName: "Science"; date: 
            "2020-09-18- 18"; volume: "369"; issue: "6510"; pages: "1501-1505"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             5486..5509
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     CDS             5513..5536
                     /codon_start=1
                     /label=8xHis
                     /note="8xHis affinity tag"
                     /translation="HHHHHHHH"
     CDS             5543..5566
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     CDS             5606..5629
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     primer_bind     complement(5731..5750)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5796..5851
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5850..5869)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(6212..6228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6236..6252)
                     /label=lac operator
                     /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(6260..6290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6305..6326)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(6455..6472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6626..7214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7388..8245)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8246..8350)
                     /label=AmpR promoter
     promoter        8376
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin 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 gggggggcgc gccrgggggg gggggggggg gggggggggg
      541 ggggggggsg rggsggagag gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1741 tcaggccgag ttcggtaccg ccaccatgtt cgtgttcctg gtgctcctgc ctctggtgag
     1801 cagccagtgc gtgaacctga ccacccgaac ccagctccca ccagcctaca ccaacagctt
     1861 tacacggggc gtgtactacc ctgacaaggt gttcagatct agcgtcctgc acagcactca
     1921 ggacctcttc ctgccgttct tcagcaacgt gacatggttc cacgccatcc acgtgagcgg
     1981 cacaaacgga accaagcggt ttgataaccc cgtcctgcca ttcaatgatg gagtttactt
     2041 cgccagtacc gagaagagta acatcatccg gggctggatc ttcggcacca ccctggatag
     2101 caaaacacag agcctcctga tcgtgaacaa tgccacgaac gtcgtgatca aggtgtgcga
     2161 gttccagttt tgcaatgatc ctttcctggg tgtgtactac cacaagaaca acaagagctg
     2221 gatggaaagc gagttcagag tctacagcag cgccaacaac tgcacattcg agtacgtctc
     2281 tcagcctttt ctgatggacc ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt
     2341 cgtgttcaag aacatcgacg gctacttcaa aatctactcc aagcacacac ccatcaacct
     2401 ggtccgggac ctccctcagg gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg
     2461 catcaacata acgcggttcc aaaccctgct ggccctgcat agatcctacc tgactcctgg
     2521 cgacagcagc agcggatgga ccgccggagc tgcagcctac tatgtgggct acctgcaacc
     2581 tagaaccttc ctgctgaagt acaacgagaa cggcacaatc acagacgccg tcgactgcgc
     2641 cctggaccct ctctctgaga caaagtgcac cctgaagtcc ttcaccgtgg aaaagggcat
     2701 ctaccagacc agcaacttcc gggtgcagcc tacagagagc atcgtgcgat ttccaaacat
     2761 taccaacctc tgccccttcg gcgaggtgtt taacgccaca agatttgcct ccgtttacgc
     2821 ctggaataga aagagaatca gcaattgtgt ggccgactac tccgtgctgt ataacagcgc
     2881 ctctttcagc accttcaagt gctacggcgt ttccccaaca aagctgaatg acctgtgctt
     2941 caccaacgtg tacgccgact ccttcgtaat tagaggcgat gaggtgcggc agatcgcacc
     3001 aggccagacc ggtaagatcg ctgactacaa ctataagctg cctgatgatt ttacaggctg
     3061 cgtgatcgcc tggaactcta acaacctgga tagcaaggtg ggcggcaact acaactacct
     3121 gtaccggctg tttcgcaagt ctaacctgaa acctttcgag agagacatct ccacagagat
     3181 ctaccaggcc ggttctacac cttgtaacgg ggtggaaggc ttcaactgtt acttccctct
     3241 gcaaagctac ggcttccagc ctaccaatgg agtcggctac cagccatacc gggtggtcgt
     3301 gctgtccttc gagttactcc acgcccccgc caccgtctgc ggtcctaaga agtccaccaa
     3361 tctggttaag aacaaatgcg tgaacttcaa cttcaacggc ctgaccggga ccggcgtgct
     3421 gaccgaaagc aacaaaaagt tcctcccctt ccagcagttc ggccgtgata tcgctgacac
     3481 cacagatgcc gtcagagatc cacagaccct ggaaatcctg gatattacac cctgctcctt
     3541 cggaggagtt tctgtgatca cccccgggac caataccagc aaccaggtgg ctgtgctgta
     3601 ccaagatgtt aactgcaccg aggttcctgt ggccatccac gccgatcagc tgacacctac
     3661 ttggagagtg tactccactg gctccaatgt gttccagacc agggccggat gtctgatcgg
     3721 cgccgagcac gtgaataaca gttacgagtg cgacatccct atcggcgccg gcatctgtgc
     3781 cagctaccag acccagacaa acagccctgg gtctgcttcc tctgtagcta gccagagcat
     3841 catcgcctac accatgagcc tgggcgcaga gaacagcgtg gcctattcca acaactctat
     3901 cgccattccc accaacttta caattagcgt cacaacagag atcctgcccg tgagcatgac
     3961 caagaccagc gtggactgta caatgtacat ctgtggcgac agcactgaat gcagcaacct
     4021 gctgctgcaa tacggctcct tttgcaccca actgaaccgg gcgctgaccg gaatcgccgt
     4081 ggaacaggac aaaaataccc aggaggtgtt cgcccaagtg aagcagatct acaagacccc
     4141 acctatcaag gacttcggcg gctttaactt tagccagatt ctccctgatc cttctaagcc
     4201 tagcaagcgg agccctatcg aggatctgct gttcaacaag gtcaccctgg ccgatgccgg
     4261 ctttatcaaa cagtatggcg attgcctggg cgacatagcc gccagagatc tgatctgcgc
     4321 ccagaaattc aacggcctga cagttctccc acctctgctg accgacgaga tgatcgctca
     4381 gtacacctct gccctgctgg ctggcaccat cacatctggg tggacatttg gcgccggccc
     4441 cgccctgcag atcccctttc ccatgcagat ggcctataga ttcaacggaa tcggcgtgac
     4501 ccagaacgtg ctgtatgaaa accagaagct gatcgctaac cagttcaatt ctgccatcgg
     4561 caagatccag gactccctct cctctacccc cagcgccctg ggcaaactgc aggacgtggt
     4621 gaatcagaac gcccaagccc tgaacaccct ggtgaagcag ctcagcagca attttggcgc
     4681 catcagctct gtgctgaacg atatcctgtc tagactggac cctccagaag ccgaagtcca
     4741 gatcgataga ctgatcacag gcagactgca gtccctgcaa acctacgtga cccaacagct
     4801 gatcagggcc gctgaaataa gagccagcgc caatctcgcc gctaccaaga tgtccgagtg
     4861 tgtgctggga cagtctaaac gcgttgactt ctgcggcaaa ggctatcacc tgatgagctt
     4921 cccccagagc gcgccgcacg gcgtggtgtt cctgcatgtg acatacgtgc ctgcccaaga
     4981 gaagaatttc acaaccgccc ctgccatctg ccacgacggc aaggcccact tcccaagaga
     5041 gggcgttttc gtttccaatg gcacacactg gttcgtgaca caaagaaact tctacgaacc
     5101 ccagattatc accaccgaca acaccttcgt gagtggcaat tgtgacgtgg tcatcggaat
     5161 cgtgaacaac acagtgtacg accctctgca acctgagctg gactctttta aggaagagct
     5221 ggacaagtac tttaaaaacc acaccagccc cgatgtggac ctgggcgaca tcagtggcat
     5281 taacgccagc gtggtgaaca tccaaaagga aatcgacaga ctgaacgagg tggccaagaa
     5341 cctgaacgag tccctgatcg acctgcagga gctcggcaaa tacgagcagg gatccggata
     5401 catccccgag gcccccagag atggccaggc ctacgtgcgg aaggacggcg agtgggtact
     5461 gctgagcaca ttcctgggca gatccctgga ggtgctgttc cagggcccag gccatcacca
     5521 ccatcaccac catcatagcg cctggtccca cccccagttc gagaagggcg gcggtagtgg
     5581 agggggcgga tctggcggct cagcttggag ccacccccag ttcgaaaagt gataatgact
     5641 cgagcgataa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     5701 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     5761 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     5821 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     5881 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     5941 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     6001 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     6061 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     6121 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     6181 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6361 gaaacctgtc gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga
     6421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8341 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
