LOCUS       sequence-356139-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for Saccharomyces cerevisiae 
            beta′-COPI-WD40 (residues 1-301)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  AUTHORS   Dey D, Singh S, Khan S, Martin M, Schnicker NJ, Gakhar L, Pierce BG,
            Hasan SS
  TITLE     An extended motif in the SARS-CoV-2 spike modulates binding and 
            release of host coatomer in retrograde trafficking.
  JOURNAL   Commun Biol. 2022 Feb 8;5(1):115. doi: 10.1038/s42003-022-03063-y.
  PUBMED    35136165
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: "Commun 
            Biol."; date: "2022-02-8"; volume: "5(1):115. doi"; pages: " 
            10.1038/s42003-022-03063-y"
COMMENT     Sequence Label: pSUMO-beta′-COPI-WD40
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(497..1579)
                     /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     1541..1560
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1580..1657)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1863..1882
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1985..2009
                     /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)."
     RBS             2024..2046
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2056..2079
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2089..2106
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2110..2400
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             3331..3348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3401..3419)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3415..3462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3499..3954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3586..3605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3796..3817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4085..4153,4154..4945)
                     /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     complement(4303..4322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5116..5704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5605..5624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5858..5875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5890..6029
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5955..5974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6115..6137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6131..6322)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
       61 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
      121 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
      181 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
      241 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
      301 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
      361 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
      421 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
      481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
      601 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
      661 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
      721 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
      781 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
      841 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
      901 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
      961 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     1021 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     1081 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     1141 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     1201 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     1261 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     1321 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     1381 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     1441 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     1501 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     1561 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     1621 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     1681 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     1741 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     1801 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     1861 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     1921 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     1981 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     2041 aggagatata ccatgtggtc acacccccaa tttgaaaaaa gtgctggtca tcaccatcat
     2101 catcacgggt cggactcaga agtcaatcaa gaagctaagc cagaggtcaa gccagaagtc
     2161 aagcctgaga ctcacatcaa tttaaaggtg tccgatggat cttcagagat cttcttcaag
     2221 atcaaaaaga ccactccttt aagaaggctg atggaagcgt tcgctaaaag acagggtaag
     2281 gaaatggact ccttaagatt cttgtacgac ggtattagaa ttcaagctga tcagacccct
     2341 gaagatttgg acatggagga taacgatatt attgaggctc accgcgaaca gattggaggt
     2401 tcgatgaagt tggatataaa gaaaactttt agtaatagat ccgatcgagt taaaggtatc
     2461 gatttccatc ctactgagcc atgggttttg actaccttgt actctggtcg tgttgagatc
     2521 tggaattacg aaacacaagt tgaagttaga tccatccaag ttacggaaac tcctgttaga
     2581 gcaggtaaat tcattgcaag aaagaattgg ataattgttg ggagtgatga tttcaggatc
     2641 agagtcttca actacaatac aggtgaaaaa gttgttgatt tcgaagctca tccagactat
     2701 atccgttcca ttgctgtaca tccaacaaag ccttatgttt tatccggcag tgatgatttg
     2761 acagtcaaac tatggaattg ggaaaataac tgggccttag aacaaacttt tgaaggtcac
     2821 gaacactttg tcatgtgcgt cgcattcaat ccaaaggacc ctagtacgtt tgcatctggg
     2881 tgtttggacc gaaccgttaa agtttggtca ttgggacaat cgacaccaaa tttcacgtta
     2941 accacaggac aagaaagagg tgtgaattat gttgattatt acccgctacc tgacaagcct
     3001 tatatgatta ctgcgtcgga cgacttaacc atcaaaattt gggattacca aacgaagtca
     3061 tgtgtagcca ctctagaagg tcacatgtcc aatgtttctt ttgctgtttt ccatccaact
     3121 ttacccatta taatttctgg ttccgaagat ggcactttaa agatatggaa ttcttccact
     3181 tacaaggtag aaaagacttt aaacgttggt ttagaaagaa gttggtgtat cgctacccat
     3241 ccaacaggac gtaaaaacta cattgcatcc gggtttgata atgggtttac cgtactttcc
     3301 ttgggatgaa agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct
     3361 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     3421 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     3481 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3541 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3601 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3661 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3721 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3781 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3841 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3901 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4081 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4141 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4201 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4261 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4321 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4381 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4441 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4501 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4561 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4621 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4681 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4741 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4801 ttctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4861 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4921 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4981 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5221 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5761 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5821 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5881 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     5941 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     6001 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     6061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     6181 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     6241 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     6301 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     6361 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     6421 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     6481 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     6541 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     6601 gcgtttccag actttacgaa acacggaaac cgaa
//
