LOCUS       sequence-436198-        6418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for Schizosaccharomyces pombe 
            alpha-COPI-WD40 mutant Arg300Ala (residues 1-327).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6418)
  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 6418)
  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-pcDNA3.1-alpha-COPI-WD40-Arg300Ala
FEATURES             Location/Qualifiers
     source          1..6418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..97
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     331..348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    465..486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(501..506,507..524,525..531)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    539..555
                     /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     544..566
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     563..579
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     563..579
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    628..749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(639..658)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     693..712
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(923..1717)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1034..1053)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1644..1663
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1784..2113)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1798..1933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1852..1871)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2098..2118
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2127..2555)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2237..2258)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2449..2468
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2601..2825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2814..2831
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2866..2889)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     regulatory      3873..3882
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3958..3977)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3959..3977)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4022..4225)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4051..4071)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4226..4605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4777..4796
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4871..4975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4976..5044,5045..5836)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5194..5213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6007..6418,1..177)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
       61 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      301 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      361 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      421 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      481 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      541 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
      601 ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa
      661 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      721 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
      781 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
      841 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
      901 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
      961 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     1021 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     1081 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     1141 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     1201 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     1261 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     1321 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     1381 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     1441 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     1501 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     1561 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     1621 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     1681 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     1741 cttgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa aagcctaggc
     1801 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg gcctcggcct
     1861 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     1921 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     1981 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     2041 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     2101 ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt
     2161 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2221 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2281 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2341 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2401 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2461 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2521 acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag agccccagct
     2581 ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc tgctattgtc
     2641 ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct
     2701 actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
     2761 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
     2821 cacagtcgag gctgatcagc gggtttaaac gggccctcta gattacttct cgaactgggg
     2881 gtggctccag gcgctctcca gcttgaacac catcacgccg ctgtcgtggg cagcggcgaa
     2941 caggttcagc ttggggtgca cggtgatgaa ccagaaggcg tcgttgtccc tcctgaaggt
     3001 ctgcacggcg gtcctcctgt tcaggtccca cactctgatg gtcttgtcct cgctggcgga
     3061 cagaatcagc tcctggtggg ggtggaacag gcagcagctc acgttgttga agtggcccct
     3121 gcaggtgtcc acctcccagg ccttgctggc ggtcatcctc cacagcttca ccagcctgtc
     3181 gtcgccggcg ctcaggatca ggggcagggt gggatggaag gcacaccagt tcacgcccct
     3241 gtcgtggccc tccagcacga acttcacgat ggcgtcggtg ctgcccttct tgtcgttgct
     3301 cttgctgttg tgggcctggg ccttctggtc ctccttgctc atgctcacag gggcggcgtt
     3361 cttcatcctc aggccgctga tgtcccacac cctcacggtc tggtccaggc tggcgctcac
     3421 gatcaggtcc tcgctggggt gaaaggcagc gcacatcacg tagtggctgt ggccggtcag
     3481 gatggcgatg cagttcctgc tctgccagtt ccagatcctg atggtctggt cgtcgctgca
     3541 gctcaggatc caggggtact cgtggtggaa ggtgcacact ctcacatagt ccatgtggcc
     3601 gcacaggctg aacagcagct tcctgctctt gtagttccac acgttcacct tgtagtcgtc
     3661 gccgccgctc acgaacaggg gctgggtggg gtggaaggcg atgcccctca cggggccgtc
     3721 gtggccgtcg aacctgtcca gcagggtgcc catcctgtag tcccacagct ggatcctgcc
     3781 gttgtgcagg ctggtcagaa tccaaggctg ggtagggtgg aaagccacgc ccttggccct
     3841 gctgctcctg ctctcgaact tggtcagcat ctccatggtg gcgaattcca ccacactgga
     3901 ctagtggatc cgagctcggt accaagctta agtttaaacg ctagccagct tgggtctccc
     3961 tatagtgagt cgtattaatt tcgataagcc agtaagcagt gggttctcta gttagccaga
     4021 gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     4081 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     4141 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     4201 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     4261 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     4321 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     4381 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     4441 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     4501 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     4561 accccgtaat tgattactat taataactag tcaataatca atgtcaacgc gtatatctgg
     4621 cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc atgcaattgt
     4681 cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca gcgacctcca
     4741 acacacaagc agggagcaga tactggctta actatgcggc atcagagcag attgtactga
     4801 gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg
     4861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgctt
//
