LOCUS       sequence-472121-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for Saccharomyces cerevisiae 
            beta′-COPI-WD40-Lys17Ala mutant (residues 1-301).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  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 6634)
  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: SSH-pSUMO-beta′-COPI-WD40-Lys17Ala
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(70..87)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1018..1308)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             complement(1312..1329)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1339..1362)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             1372..1394
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1409..1433
                     /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(1433..1452)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1434..1452)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1536..1555)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1761..1838
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1839..2921
                     /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(1858..1877)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3730..3921
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3915..3937)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4023..4162
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4078..4097
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4177..4194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4348..4936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4428..4447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5107..5898,5899..5967))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5730..5749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5968..6072)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6098..6553)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6235..6256)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6447..6466
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(join(6590..6634,1..3))
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     join(6633..6634,1..17)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc
       61 cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagctttc atcccaagga
      121 aagtacggta aacccattat caaacccgga tgcaatgtag tttttacgtc ctgttggatg
      181 ggtagcgata caccaacttc tttctaaacc aacgtttaaa gtcttttcta ccttgtaagt
      241 ggaagaattc catatcttta aagtgccatc ttcggaacca gaaattataa tgggtaaagt
      301 tggatggaaa acagcaaaag aaacattgga catgtgacct tctagagtgg ctacacatga
      361 cttcgtttgg taatcccaaa ttttgatggt taagtcgtcc gacgcagtaa tcatataagg
      421 cttgtcaggt agcgggtaat aatcaacata attcacacct ctttcttgtc ctgtggttaa
      481 cgtgaaattt ggtgtcgatt gtcccaatga ccaaacttta acggttcggt ccaaacaccc
      541 agatgcaaac gtactagggt cctttggatt gaatgcgacg cacatgacaa agtgttcgtg
      601 accttcaaaa gtttgttcta aggcccagtt attttcccaa ttccatagtt tgactgtcaa
      661 atcatcactg ccggataaaa cataaggctt tgttggatgt acagcaatgg aacggatata
      721 gtctggatga gcttcgaaat caacaacttt ttcacctgta ttgtagttga agactctgat
      781 cctgaaatca tcactcccaa caattatcca attctttctt gcaatgaatt tacctgctct
      841 aacaggagtt tccgtaactt ggatggatct aacttcaact tgtgtttcgt aattccagat
      901 ctcaacacga ccagagtaca aggtagtcaa aacccatggc tcagtaggat ggaaatcgat
      961 accagcaact cgatcggatc tattactaaa agttttcttt atatccaact tcatcgaacc
     1021 tccaatctgt tcgcggtgag cctcaataat atcgttatcc tccatgtcca aatcttcagg
     1081 ggtctgatca gcttgaattc taataccgtc gtacaagaat cttaaggagt ccatttcctt
     1141 accctgtctt ttagcgaacg cttccatcag ccttcttaaa ggagtggtct ttttgatctt
     1201 gaagaagatc tctgaagatc catcggacac ctttaaattg atgtgagtct caggcttgac
     1261 ttctggcttg acctctggct tagcttcttg attgacttct gagtccgacc cgtgatgatg
     1321 atggtgatga ccagcacttt tttcaaattg ggggtgtgac cacatggtat atctccttct
     1381 taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag
     1441 tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc
     1501 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     1561 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
     1621 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
     1681 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
     1741 gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     1801 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     1861 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     1921 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     1981 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     2041 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     2101 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     2161 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     2221 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     2281 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     2341 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     2401 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     2461 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     2521 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     2581 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     2641 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     2701 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     2761 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     2821 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2881 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
     2941 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca
     3001 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     3061 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     3121 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     3181 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     3241 attatcgccg gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
     3301 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
     3361 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
     3421 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
     3481 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     3541 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
     3601 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
     3661 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
     3721 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     3781 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     3841 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
     3901 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3961 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4021 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     4081 catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt
     4141 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4861 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5221 tggccccagt gctgcaatga taccgcgaga accacgctca ccggctccag atttatcagc
     5281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5401 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     6121 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6181 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6241 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6301 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6361 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6421 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6481 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6541 ctacagggcg cgtcccattc gccaatccgg atatagttcc tcctttcagc aaaaaacccc
     6601 tcaagacccg tttagaggcc ccaaggggtt atgc
//
