LOCUS       sequence-436194-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for the cytosolic tail of SARS-CoV-2 
            spike (with Cys1254Ala and Thr1273Glu substitutions) and an 
            N-terminal eXact tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Dey D, Qing E, He Y, Chen Y, Jennings B, Cohn W, Singh S, Gakhar L, 
            Schnicker NJ, Pierce BG, Whitelegge JP, Doray B, Orban J, Gallagher 
            T, Hasan SS
  TITLE     A single C-terminal residue controls SARS-CoV-2 spike trafficking 
            and incorporation into VLPs.
  JOURNAL   Nat Commun. 2023 Dec 15;14(1):8358. doi: 10.1038/s41467-023-44076-3.
  PUBMED    38102143
REFERENCE   2  (bases 1 to 6084)
  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: "2023-12-15"; volume: "14(1):8358. doi"; pages: " 
            10.1038/s41467-023-44076-3"
COMMENT     Sequence Label: JO-pH720-SARS-CoV-2-S-tail-Cys1254Ala-Thr1273Glu
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        134..211
                     /gene="lacI"
                     /label=lacI promoter
     CDS             212..1294
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(231..250)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2606..2797
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2791..2813)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2899..3038
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2954..2973
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3053..3070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3224..3812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3304..3323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3983..4774,4775..4843))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4606..4625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4844..4948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5016..5034
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5061..5066,5067..5083,5084..5089)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(5264..5311)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5307..5325
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             5829..5851
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5866..5890
                     /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)."
     primer_bind     complement(5890..5909)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5891..5909)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5993..6012)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
       61 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
      121 cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
      181 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
      241 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
      301 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
      361 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
      421 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
      481 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
      541 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
      601 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
      661 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
      721 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
      781 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
      841 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
      901 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
      961 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     1021 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     1081 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     1141 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     1201 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1261 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     1321 tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct
     1381 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     1441 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     1501 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     1561 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     1621 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
     1681 tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     1741 ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg
     1801 ctcttaccag cctaacttcg atcattggac cgctgatcgt cacggcgatt tatgccgcct
     1861 cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc
     1921 tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg
     1981 gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac
     2041 tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag
     2101 ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct
     2161 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     2221 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     2281 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     2341 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     2401 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     2461 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     2521 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     2581 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     2641 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     2701 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     2761 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2821 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2881 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     2941 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg cggtgtgaaa
     3001 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctgttccgct tcctcgctca
     3061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3661 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3901 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3961 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4021 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4081 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4141 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4201 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4261 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     4321 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4381 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4441 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4501 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4561 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     4621 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4681 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4741 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4801 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4921 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4981 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5041 gtcttcaaga aatctcatgt ttgacagctt atcatcgata ggctttaatg cggtagttta
     5101 tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc
     5161 gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg
     5221 ccgggcctct tgcgggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
     5281 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
     5341 cagcttcctt tcgggcttat gcggccgcct cgaggaattc ttattcgtag tgcagcttca
     5401 cgcccttcag cacgggctcg ctgtcgtcct cgtcgaactt gcacgccagg ccgcggtact
     5461 ggtggtcaaa ttcaactttt tcaacgcccg gcattttctt cagtttgcca actgcattgg
     5521 ccggcacatc cacgacaacg gccgggatca gtttgaactg atagacgatg tgaccgccaa
     5581 tacccagaac ttcatgcggg ttaaatttcg ctttatccac gctcacgatc acgcgaatcg
     5641 tattgcctga gccttccgtc ggcagtgctt tcaggatttc atttttcaga ctttcaacgc
     5701 cttccaccgt tttcgcattg ttgatcagtt taatgtattt gtcaccgatg ccgtactgtt
     5761 tcagttcttt aatagctgct tctttggcct gcgccagaga attggcatcc accgcttcca
     5821 tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc
     5881 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac
     5941 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     6001 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     6061 ggcgtgggta tggtggcagg cccc
//
