LOCUS       sequence-407417-        8529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human SARS coronavirus S protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8529)
  AUTHORS   Strebinger D, Frangieh CJ, Friedrich MJ, Faure G, Macrae RK, Zhang F
  TITLE     Cell type-specific delivery by modular envelope design.
  JOURNAL   Nat Commun. 2023 Aug 23;14(1):5141. doi: 10.1038/s41467-023-40788-8.
  PUBMED    37612276
REFERENCE   2  (bases 1 to 8529)
  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: "Nat 
            Commun."; date: "2023-08-23"; volume: "14(1):5141. doi"; pages: " 
            10.1038/s41467-023-40788-8"
COMMENT     Sequence Label: pHCMV_SARS-CoV_S
FEATURES             Location/Qualifiers
     source          1..8529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    5139..5150
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     primer_bind     complement(5234..5253)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5299..5354
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5353..5372)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5712..5728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5712..5728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5725..5747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5736..5752
                     /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(5760..5766,5767..5784,5785..5790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5805..5826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5943..5960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6114..6702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6194..6213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6873..7664,7665..7733))
                     /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     7496..7515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7734..7838)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7864..8319)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8001..8022)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8213..8232
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8446..8468
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8460..8477
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8461..8477
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8487..8506
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8487..8505
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaca cgccatccac
      721 gctgttttga cctccataga tgacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattcgc caccatgttc atctttctgc
     1441 tgtttctgac actgacctcc ggctccgatc tcgatagatg caccaccttc gacgacgtgc
     1501 aagcccctaa ttacacacag cacaccagca gcatgagggg agtctactac cccgacgaaa
     1561 tcttcagaag cgatacactg tatctgacac aagatttgtt tctgcccttc tactccaacg
     1621 tcaccggatt tcacaccatc aatcacacct tcggcaaccc cgtgattccc ttcaaagacg
     1681 gaatttattt cgctgccacc gaaaaaagca atgtggtgag aggatgggtg ttcggctcca
     1741 caatgaacaa caaaagccaa tccgtgatca tcattaacaa ttccaccaat gtggtgatta
     1801 gagcttgcaa cttcgaactg tgcgataacc ccttcttcgc tgtgtccaaa cccatgggca
     1861 cacagaccca caccatgatc ttcgacaatg cctttaattg cacatttgag tatatctccg
     1921 acgctttctc tctggatgtc agcgaaaaga gcggcaactt caagcatctg agagagttcg
     1981 tgttcaaaaa taaggacgga tttctgtacg tgtacaaggg atatcagccc atcgacgtgg
     2041 tgagggatct cccttccggc ttcaacacac tgaagcctat cttcaagctc cccctcggca
     2101 tcaatatcac caattttagg gctattctca ccgcctttag ccccgcccaa gacatttggg
     2161 gcacaagcgc cgctgcctat ttcgtgggct atctgaagcc caccaccttt atgctgaaat
     2221 acgacgaaaa cggcacaatc accgatgctg tcgactgcag ccagaaccct ctcgccgagc
     2281 tcaagtgtag cgtgaaatcc tttgaaatcg ataagggaat ctaccagacc agcaacttta
     2341 gggtggtccc cagcggcgac gtggtgaggt tccccaacat taccaatctc tgccccttcg
     2401 gagaggtgtt caacgccaca aaattcccca gcgtgtacgc ttgggagaga aagaaaattt
     2461 ccaattgcgt cgctgactac agcgtgctgt acaactccac cttcttttcc accttcaagt
     2521 gctacggagt gtccgccaca aagctgaatg atctgtgctt ctccaacgtg tacgccgaca
     2581 gctttgtggt caagggagat gatgtgaggc aaatcgctcc cggccagacc ggcgtcatcg
     2641 ctgattacaa ctacaaactg cccgacgact tcatgggatg cgtgctcgct tggaatacca
     2701 gaaatattga tgctacatcc accggcaact acaactacaa gtatagatat ctgagacacg
     2761 gcaaactgag acctttcgag agggacatca gcaatgtccc tttcagcccc gacggaaagc
     2821 cttgcacccc tcccgctctg aattgctatt ggcctctgaa cgactacggc ttctacacca
     2881 caaccggcat tggctatcag ccctacagag tcgtggtgct gagcttcgaa ctgctcaatg
     2941 cccccgccac cgtctgtggc cctaaactct ccacagacct cattaagaat caatgcgtca
     3001 acttcaactt caatggactg accggaaccg gcgtgctgac cccttccagc aagagatttc
     3061 agcccttcca gcagtttgga agggacgtgt ccgacttcac agacagcgtg agggatccca
     3121 aaacctccga gattctcgac atctcccctt gcgcttttgg cggagtctcc gtgatcaccc
     3181 ccggaaccaa tgctagctcc gaagtggctg tgctgtacca agacgtgaac tgtaccgacg
     3241 tgtccaccgc cattcacgct gaccaactca cacccgcttg gagaatctac tccaccggca
     3301 ataatgtgtt ccagacacaa gccggatgtc tgattggcgc cgagcacgtc gacacaagct
     3361 acgagtgcga catccctatt ggagccggaa tctgtgccag ctaccacaca gtgtctctgc
     3421 tgaggtccac cagccaaaaa agcatcgtcg cctacaccat gagcctcgga gccgactcct
     3481 ccattgccta ctccaataac accatcgcta tccccacaaa ttttagcatc agcatcacca
     3541 ccgaggtcat gcccgtgtcc atggctaaga ccagcgtgga ctgcaacatg tatatctgcg
     3601 gcgattccac cgaatgtgcc aatctgctgc tgcaatacgg ctccttttgc acccagctga
     3661 acagagccct cagcggaatc gctgccgagc aagatagaaa cacaagggag gtgttcgccc
     3721 aagtgaagca gatgtataag acccccacac tgaagtattt cggaggcttt aactttagcc
     3781 aaattctccc cgaccctctg aaacccacca aaaggagctt tattgaggat ctgctgttta
     3841 ataaagtgac cctcgccgat gccggcttta tgaagcaata tggcgagtgc ctcggagaca
     3901 ttaacgctag ggatctgatc tgtgcccaga agtttaacgg actgacagtg ctgccccctc
     3961 tgctcacaga cgacatgatc gctgcctaca cagctgctct ggtgtccggc acagctacag
     4021 ccggctggac cttcggcgcc ggcgccgccc tccaaatccc cttcgccatg cagatggcct
     4081 atagattcaa cggcattggc gtgacacaaa acgtgctgta cgagaatcag aaacaaatcg
     4141 ctaaccagtt taataaggcc atcagccaga ttcaagagtc cctcacaacc acaagcacag
     4201 ctctgggcaa gctgcaagac gtcgtcaatc aaaacgctca agctctgaac accctcgtga
     4261 agcagctgtc cagcaacttt ggagccatca gcagcgtcct caacgacatt ctgtccagac
     4321 tcgataaggt ggaggccgag gtgcagatcg ataggctgat caccggcaga ctgcaatctc
     4381 tgcaaaccta tgtgacccag caactcatca gagccgccga gatcagagcc agcgccaatc
     4441 tggctgctac aaaaatgagc gaatgcgtcc tcggacagag caaaagagtg gatttctgcg
     4501 gcaagggata ccatctgatg tcctttcccc aagctgcccc tcacggagtg gtctttctcc
     4561 acgtgacata cgtgccttcc caagagagaa acttcaccac cgctcccgct atctgccatg
     4621 agggcaaggc ctattttccc agagaaggcg tctttgtgtt caacggaacc agctggttta
     4681 tcacccagag aaacttcttt tccccccaga tcattaccac cgataatacc ttcgtgagcg
     4741 gcaattgtga tgtggtgatc ggaatcatta ataacacagt ctacgatcct ctgcagcccg
     4801 agctggatag cttcaaggaa gagctggaca agtacttcaa gaaccatacc agccccgatg
     4861 tcgatctcgg cgatatctcc ggcatcaacg ccagcgtggt gaacattcaa aaagagatcg
     4921 atagactgaa tgaagtcgcc aaaaatctga atgaaagcct cattgacctc caagaactcg
     4981 gcaagtacga acaatacatc aagtggcctt ggtacgtgtg gctgggcttt atcgctggac
     5041 tgatcgctat cgtgatggtc accattctgc tgtgctgcat gaccagctgc tgcagctgtc
     5101 tgaagggcgc ttgcagctgc ggcagctgtt gcaagttcga tgaggatgac agcgagcccg
     5161 tgctgaaggg cgtcaagctc cattatacct gatgtcttcc tggagctcca gtcattgact
     5221 ataacgtctc agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     5281 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     5341 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     5401 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     5461 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     5521 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     5581 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     5641 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacgga tcggccgcaa
     5701 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5761 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5821 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5881 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5941 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6001 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6061 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6121 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6181 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6241 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6301 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6361 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6421 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6481 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6541 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6601 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6661 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6721 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6781 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6841 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6901 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6961 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7021 cgctcaccgg caccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7081 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7141 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7201 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7261 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7321 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7381 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7441 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7501 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7561 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7621 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7681 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7741 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7801 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7861 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     7921 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     7981 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     8041 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     8101 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     8161 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     8221 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     8281 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     8341 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     8401 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     8461 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctcc
     8521 accgcggtg
//
