LOCUS       sequence-466904-        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for the SARS-CoV-2 cytosolic tail 
            (Thr1273Glu 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-Thr1273Glu
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             178..189
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(366..388)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(533..551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        619..723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(724..792,793..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     complement(942..961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1755..2343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2244..2263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2497..2514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2587..2664
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2665..3747
                     /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(2684..2703)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3796..3801,3802..3819,3820..3826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3834..3850
                     /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     3858..3874
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3858..3874
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3870..4397
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3890..3907)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3890..3906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3899..3921)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(4499..4504,4505..4520,4521..4527)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4535..4551
                     /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             join(4574..5059,1..168)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
ORIGIN
        1 ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt
       61 attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct
      121 ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggatctgatc
      181 gaaggtcgtg ggatccccag gaattccctc aagggctgtt gttcttgtgg atcctgctgc
      241 aaatttgatg aagacgactc tgagccagtg ctcaaaggag tcaaattaca ttacgagtaa
      301 ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt
      361 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      421 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
      481 atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg
      541 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
      601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      721 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      781 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      841 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      901 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      961 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1021 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1081 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1141 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1201 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1261 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1321 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1381 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1441 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1501 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1561 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1621 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1681 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1741 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1801 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1861 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     1921 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1981 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2041 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2101 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2161 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2221 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2281 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2341 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2401 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2461 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2521 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2581 aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     2641 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     2701 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     2761 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     2821 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     2881 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     2941 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     3001 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     3061 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     3121 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     3181 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     3241 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     3301 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     3361 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     3421 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     3481 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     3541 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     3601 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     3661 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3721 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     3781 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     3841 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
     3901 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3961 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4021 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
     4081 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
     4141 ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca
     4201 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
     4261 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaattagct
     4321 tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat
     4381 ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg
     4441 gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt
     4501 gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     4561 ggaaacagta ttcatgtccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc
     4621 cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga
     4681 tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc
     4741 ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc
     4801 tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga
     4861 aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga
     4921 aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg
     4981 tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta
     5041 tgacgctctt gatgttgtt
//
