LOCUS       sequence-436201-        6712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for Schizosaccharomyces pombe 
            alpha-COPI-WD40.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6712)
  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 6712)
  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-alpha-COPI-WD40
FEATURES             Location/Qualifiers
     source          1..6712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             614..805
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(799..821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    907..1046
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     962..981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1061..1078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1232..1820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1312..1331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1991..2782,2783..2851))
                     /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     2614..2633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2852..2956)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2982..3437)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3119..3140)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3331..3350
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3474..3521)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3517..3535
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3588..3605)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4614..4904)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             complement(4908..4925)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4935..4958)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             4968..4990
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5005..5029
                     /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(5029..5048)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5030..5048)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5132..5151)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5357..5434
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5435..6517
                     /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(5454..5473)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
       61 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
      121 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag
      181 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
      241 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
      301 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
      361 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
      421 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
      481 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
      541 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
      601 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
      661 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
      721 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
      781 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      841 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      901 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
      961 gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat
     1021 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     1081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     1681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2101 catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat
     2161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2281 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     2341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     3001 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3061 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3121 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3181 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3241 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3301 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3361 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3421 gccgctacag ggcgcgtccc attcgccaat ccggatatag ttcctccttt cagcaaaaaa
     3481 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
     3541 gcagccaact cagcttcctt tcgggctttg ttagcagccg gatctcagtg gtggtggtgg
     3601 tggtgctcga gtgcggccgc aagcttttac tccagcttga acaccatcac gccgctgtcg
     3661 tgggcagcgg cgaacaggtt cagcttgggg tgcacggtga tgaaccagaa cctgtcgttg
     3721 tccctcctga aggtctgcac ggcggtcctc ctgttcaggt cccacactct gatggtcttg
     3781 tcctcgctgg cggacagaat cagctcctgg tgggggtgga acaggcagca gctcacgttg
     3841 ttgaagtggc ccctgcaggt gtccacctcc caggccttgc tggcggtcat cctccacagc
     3901 ttcaccagcc tgtcgtcgcc ggcgctcagg atcaggggca gggtgggatg gaaggcacac
     3961 cagttcacgc ccctgtcgtg gccctccagc acgaacttca cgatggcgtc ggtgctgccc
     4021 ttcttgtcgt tgctcttgct gttgtgggcc tgggccttct ggtcctcctt gctcatgctc
     4081 acaggggcgg cgttcttcat cctcaggccg ctgatgtccc acaccctcac ggtctggtcc
     4141 aggctggcgc tcacgatcag gtcctcgctg gggtgaaagg cagcgcacat cacgtagtgg
     4201 ctgtggccgg tcaggatggc gatgcagttc ctgctctgcc agttccagat cctgatggtc
     4261 tggtcgtcgc tgcagctcag gatccagggg tactcgtggt ggaaggtgca cactctcaca
     4321 tagtccatgt ggccgcacag gctgaacagc agcttcctgc tcttgtagtt ccacacgttc
     4381 accttgtagt cgtcgccgcc gctcacgaac aggggctggg tggggtggaa ggcgatgccc
     4441 ctcacggggc cgtcgtggcc gtcgaacctg tccagcaggg tgcccatcct gtagtcccac
     4501 agctggatcc tgccgttgtg caggctggtc agaatccaag gctgggtagg gtggaaagcc
     4561 acgcccttgg ccctgctgct cctgctctcg aacttggtca gcatctccat cgaacctcca
     4621 atctgttcgc ggtgagcctc aataatatcg ttatcctcca tgtccaaatc ttcaggggtc
     4681 tgatcagctt gaattctaat accgtcgtac aagaatctta aggagtccat ttccttaccc
     4741 tgtcttttag cgaacgcttc catcagcctt cttaaaggag tggtcttttt gatcttgaag
     4801 aagatctctg aagatccatc ggacaccttt aaattgatgt gagtctcagg cttgacttct
     4861 ggcttgacct ctggcttagc ttcttgattg acttctgagt ccgacccgtg atgatgatgg
     4921 tgatgaccag cacttttttc aaattggggg tgtgaccaca tggtatatct ccttcttaaa
     4981 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
     5041 tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc
     5101 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     5161 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     5221 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     5281 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     5341 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     5401 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     5461 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     5521 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     5581 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     5641 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     5701 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     5761 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     5821 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     5881 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     5941 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     6001 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     6061 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     6121 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     6181 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     6241 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     6301 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     6361 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     6421 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6481 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     6541 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
     6601 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     6661 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc ga
//
