LOCUS       sequence-432648-        6418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for Schizosaccharomyces pombe 
            alpha-COPI-WD40 Arg13Ala mutant (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-Arg13Ala
FEATURES             Location/Qualifiers
     source          1..6418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      545..1133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1034..1053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1287..1304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1421..1442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1457..1462,1463..1480,1481..1487)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1495..1511
                     /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     1500..1522
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1519..1535
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1519..1535
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1584..1705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1595..1614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1649..1668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1879..2673)
                     /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(1990..2009)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2600..2619
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2740..3069)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2754..2889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2808..2827)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3054..3074
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3083..3511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3193..3214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3405..3424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3557..3781
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3770..3787
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3822..3845)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     regulatory      4829..4838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4914..4933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4915..4933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4978..5181)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5007..5027)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5182..5561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5733..5752
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5827..5931
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5932..6000,6001..6418,1..374)
                     /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(6150..6169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
       61 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      121 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      181 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      241 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      301 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      361 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      421 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      481 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      541 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1141 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1201 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1261 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1321 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1381 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     1441 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     1501 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag
     1561 aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1621 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1681 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
     1741 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     1801 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     1861 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     1921 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     1981 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     2041 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     2101 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     2161 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     2221 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     2281 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     2341 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     2401 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     2461 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     2521 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     2581 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     2641 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     2701 atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc
     2761 aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc
     2821 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     2881 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     2941 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     3001 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     3061 ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3121 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3181 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3241 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3301 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3361 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3421 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3481 gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt
     3541 ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc
     3601 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     3661 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     3721 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     3781 gtcgaggctg atcagcgggt ttaaacgggc cctctagatt acttctcgaa ctgggggtgg
     3841 ctccaggcgc tctccagctt gaacaccatc acgccgctgt cgtgggcagc ggcgaacagg
     3901 ttcagcttgg ggtgcacggt gatgaaccag aacctgtcgt tgtccctcct gaaggtctgc
     3961 acggcggtcc tcctgttcag gtcccacact ctgatggtct tgtcctcgct ggcggacaga
     4021 atcagctcct ggtgggggtg gaacaggcag cagctcacgt tgttgaagtg gcccctgcag
     4081 gtgtccacct cccaggcctt gctggcggtc atcctccaca gcttcaccag cctgtcgtcg
     4141 ccggcgctca ggatcagggg cagggtggga tggaaggcac accagttcac gcccctgtcg
     4201 tggccctcca gcacgaactt cacgatggcg tcggtgctgc ccttcttgtc gttgctcttg
     4261 ctgttgtggg cctgggcctt ctggtcctcc ttgctcatgc tcacaggggc ggcgttcttc
     4321 atcctcaggc cgctgatgtc ccacaccctc acggtctggt ccaggctggc gctcacgatc
     4381 aggtcctcgc tggggtgaaa ggcagcgcac atcacgtagt ggctgtggcc ggtcaggatg
     4441 gcgatgcagt tcctgctctg ccagttccag atcctgatgg tctggtcgtc gctgcagctc
     4501 aggatccagg ggtactcgtg gtggaaggtg cacactctca catagtccat gtggccgcac
     4561 aggctgaaca gcagcttcct gctcttgtag ttccacacgt tcaccttgta gtcgtcgccg
     4621 ccgctcacga acaggggctg ggtggggtgg aaggcgatgc ccctcacggg gccgtcgtgg
     4681 ccgtcgaacc tgtccagcag ggtgcccatc ctgtagtccc acagctggat cctgccgttg
     4741 tgcaggctgg tcagaatcca aggctgggta gggtggaaag ccacgccctt ggcagcgctg
     4801 ctcctgctct cgaacttggt cagcatctcc atggtggcga attccaccac actggactag
     4861 tggatccgag ctcggtacca agcttaagtt taaacgctag ccagcttggg tctccctata
     4921 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     4981 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     5041 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     5101 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     5161 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     5221 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     5281 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     5341 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     5401 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5461 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     5521 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     5581 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     5641 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     5701 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     5761 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     5821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5941 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6001 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6061 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6121 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6181 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6241 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6301 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6361 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgtt
//
