LOCUS       sequence-436203-        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for the SARS-CoV-2 cytosolic tail 
            (Thr1273Asp substitution) with an N-terminal GST tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5059)
  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 5059)
  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: BD-pGEX5X3-BD-SARS-CoV-2-S-tail-Thr1273Asp
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      488..1076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     977..996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1230..1247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1320..1397
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1398..2480
                     /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(1417..1436)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2529..2534,2535..2552,2553..2559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2567..2583
                     /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     2591..2607
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2591..2607
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2603..3130
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2623..2640)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2623..2639)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2632..2654)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(3232..3237,3238..3253,3254..3260)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3268..3284
                     /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)."
     CDS             3307..3960
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3918..3940
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3970..3981
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4158..4180)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4325..4343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4411..4515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4516..4584,4585..5059,1..317)
                     /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     complement(4734..4753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
       61 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      121 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      181 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      241 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      301 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      601 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1201 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1261 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
     1321 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     1381 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     1441 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     1501 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     1561 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     1621 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     1681 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     1741 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     1801 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     1861 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     1921 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     1981 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     2041 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     2101 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     2161 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     2221 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     2281 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     2341 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     2401 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2461 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     2521 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2581 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     2641 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2701 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2761 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
     2821 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     2881 gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc
     2941 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     3001 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac
     3061 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
     3121 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
     3181 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt
     3241 aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca
     3301 gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     3361 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     3421 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     3481 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     3541 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     3601 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     3661 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     3721 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     3781 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     3841 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     3901 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     3961 tcggatctga tcgaaggtcg tgggatcccc aggaattccc tcaagggctg ttgttcttgt
     4021 ggatcctgct gcaaatttga tgaagacgac tctgagccag tgctcaaagg agtcaaatta
     4081 cattacgact aactcgagcg gccgcatcgt gactgactga cgatctgcct cgcgcgtttc
     4141 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4201 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4261 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtataattc ttgaagacga
     4321 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4381 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4741 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     4801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5041 cgtgacacca cgatgcctg
//
