LOCUS       sequence-436193-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for the cytosolic tail of SARS-CoV-2 
            spike with 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-WT
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(724..1515,1516..1584))
                     /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     1347..1366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1585..1689)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1757..1775
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1802..1807,1808..1824,1825..1830)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2005..2052)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2048..2066
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             2570..2592
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2607..2631
                     /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(2631..2650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2632..2650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2734..2753)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2959..3036
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3037..4119
                     /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(3056..3075)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5431..5622
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5616..5638)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5724..5863
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5779..5798
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5878..5895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6049..6084,1..553))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
       61 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      121 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      181 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      241 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      301 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      361 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
      421 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      481 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      541 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
      601 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
      661 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      721 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      781 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
      841 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
      901 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
      961 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1021 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1081 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1141 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1201 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1261 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1321 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     1381 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1441 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1501 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1561 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1681 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     1741 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aaatctcatg
     1801 tttgacagct tatcatcgat aggctttaat gcggtagttt atcacagtta aattgctaac
     1861 gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca
     1921 ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata
     1981 tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag
     2041 gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggctta
     2101 tgcggccgcc tcgaggaatt cttaggtgta gtgcagcttc acgcccttca gcacgggctc
     2161 gctgtcgtcc tcgtcgaact tgcagcacag gccgcggtac tggtggtcaa attcaacttt
     2221 ttcaacgccc ggcattttct tcagtttgcc aactgcattg gccggcacat ccacgacaac
     2281 ggccgggatc agtttgaact gatagacgat gtgaccgcca atacccagaa cttcatgcgg
     2341 gttaaatttc gctttatcca cgctcacgat cacgcgaatc gtattgcctg agccttccgt
     2401 cggcagtgct ttcaggattt catttttcag actttcaacg ccttccaccg ttttcgcatt
     2461 gttgatcagt ttaatgtatt tgtcaccgat gccgtactgt ttcagttctt taatagctgc
     2521 ttctttggcc tgcgccagag aattggcatc caccgcttcc atatgtatat ctccttctta
     2581 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
     2641 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg
     2701 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     2761 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     2821 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
     2881 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     2941 gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     3001 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     3061 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     3121 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     3181 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     3241 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     3301 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     3361 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     3421 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     3481 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     3541 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     3601 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     3661 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     3721 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     3781 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     3841 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     3901 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     3961 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     4021 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4081 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
     4141 ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt
     4201 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     4261 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     4321 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     4381 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     4441 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     4501 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     4561 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     4621 cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc
     4681 cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt
     4741 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     4801 cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg
     4861 agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc
     4921 agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     4981 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     5041 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     5101 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
     5161 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     5221 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
     5281 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
     5341 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
     5401 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
     5461 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     5521 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     5581 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5641 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5701 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     5761 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg
     5821 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctgtt ccgcttcctc
     5881 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5941 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6001 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6061 ccgcccccct gacgagcatc acaa
//
