LOCUS       sequence-202559-        5646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Mup1 with Mup1-iYFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5646)
  AUTHORS   Day KJ, Casler JC, Glick BS
  TITLE     Budding Yeast Has a Minimal Endomembrane System.
  JOURNAL   Dev Cell. 2018 Jan 8;44(1):56-72.e4. doi: 
            10.1016/j.devcel.2017.12.014. Epub 2018 Jan 8.
  PUBMED    29316441
REFERENCE   2  (bases 1 to 5646)
  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"; doi: 
            "10.1016/j.devcel.2017.12.014"; journalName: "Dev Cell"; date: 
            "2018-01-8- 8"; volume: "44"; issue: "1"; pages: "56-72.e4"
COMMENT     Sequence Label: YIplac211-MUP1-iYFP
FEATURES             Location/Qualifiers
     source          1..5646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(943..971)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(2144..2161)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2144..2160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2153..2175)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2369..2388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2488..2510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2628..3431)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3432..3647)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3654..3672)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3740..3844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3845..3913,3914..4705)
                     /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(4063..4082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4876..5464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5365..5384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5618..5635
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gacgtgaagg tgtgttgccc ttttctaatt tctttgcctc ttctaagcca
      301 ttcaactccc caatggttgg tttgtttcaa cattttattg tttgtacagt aaccatttta
      361 gccccacctc caggtgatgc atatttgcta gtccaaaatt taatttccta cccaatgaat
      421 attattaatt ttgccattag cgcgggtttg ctttggatat actggcaacg tagacaaggt
      481 aaaattgaat ggaacccacc aattaaggcc ggtgtttttg ttacgggatt tttcacatta
      541 tccaatttat acctgattat tgctccttac gttccacctt caaatggcga gtcggtgtat
      601 tcaagcatgc catactggat ccactgtgta attgcttggg gtattttctt ctttggtggt
      661 gtttactacg ttgtttgggc ccaattattg ccaagatggg gccactacaa attggtttcc
      721 aaggatgtgc ttggtgaaga tgggttctgg agagtcaaaa ttgccaaagt ttatgatgac
      781 actattggcg atgtcgatac acaagaagac ggcgttattg aaacgaatat aatcgaacat
      841 tacaaaagtg aacaagaaaa atcgctgaaa gatccaggaa gttcggggtc cagcggttct
      901 tcaggcagtt ccatggatag cactgagagc cttttcactg gagttgtccc aattcttgtt
      961 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     1021 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcct
     1081 tggccaacac ttgtcactac tttaggttat ggtttgcagt gctttgcaag atacccggat
     1141 catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
     1201 actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt
     1261 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
     1321 cttgggcaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
     1381 caaaagaatg gaatcaaagc taacttcaaa attagacaca acattgaaga tggaagcgtt
     1441 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     1501 gacaaccatt acctgtccta tcaatctgcc ctttcgaaag atcccaacga aaagagagac
     1561 cacatggtcc ttaaagagtt tgtaacagct gctgggatta cacatggcag ctccggctcc
     1621 agcgggtaaa gcggccgccc aaaaaggtac ttaccatctc atctcgattc ttataatcac
     1681 gtatgaacat tcttataatt gcattttctt tatacaaaat atataggtac caaacctcat
     1741 tataatttct gccatttaat agtttaagat gctattccgt ttcgtatttt ttcaattttt
     1801 atacctacta ttgcgcgaaa tcaaaatagc atactaagaa aaagcaatat catgaattgc
     1861 tgatgagata gattcaaatt ggaattaact aaatctatca gcaacgctat aaacacagat
     1921 aagcgctcta taaaatcgtt tgtctaacgt tttatatagt tctttattgc tcccttaccc
     1981 tctttccgtc aatagtgatt gttgtctcgg gaaagctttt ttttttatta atcaaaggcc
     2041 caattgagtt acaactgaaa ttaagctgag ttggcggaac agtgtggtat tagcgaaggg
     2101 aaaatctgtg aagtgcccta gctacaaaac agagctcgaa ttcactggcc gtcgttttac
     2161 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2221 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2281 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2341 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2401 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2461 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2521 catcaccgaa acgcgcgaga cgaaagggcg ggtaataact gatataatta aattgaagct
     2581 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
     2641 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
     2701 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     2761 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     2821 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     2881 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     2941 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     3001 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     3061 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     3121 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     3181 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     3241 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     3301 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     3361 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     3421 gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag ttgggttaag
     3481 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     3541 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     3601 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctggcctcgt
     3661 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3721 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3841 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3901 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3961 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4021 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4081 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4141 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4201 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4261 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4321 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4381 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4441 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4501 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4561 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4621 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4681 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4741 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5581 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5641 ggaaga
//
