LOCUS       sequence-202547-        6946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Chs3 with Chs3-msGFPx3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  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 6946)
  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-CHS3-msGFPx3
FEATURES             Location/Qualifiers
     source          1..6946
                     /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(3444..3461)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3444..3460)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3453..3475)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3669..3688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3788..3810
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3928..4731)
                     /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(4732..4947)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4954..4972)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5040..5144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5145..5213,5214..6005)
                     /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(5363..5382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6176..6764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6665..6684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6918..6935
                     /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 ccaagctttt
      241 gaatctgtct tcggttcggt aacttgtttg ccgggatgtt tctcaatgta tcgtataaaa
      301 tctcctaaag gttcagatgg ttattgggta cctgtattgg caaatccaga tattgttgaa
      361 agatattcgg ataatgtcac aaacactttg cataagaaga acttattatt acttggtgaa
      421 gatagatttt tatcttcatt aatgttaaag actttcccta agagaaagca agtatttgtt
      481 ccaaaagctg cttgtaaaac tattgcccct gataaattca aagtcttact ttcccagcgt
      541 cgaagatgga ttaattctac ggtacataac ctttttgaat tagttctaat cagagactta
      601 tgtggcactt tctgtttttc catgcaattt gtgattggta ttgaattgat tggtactatg
      661 gtactgccgt tagccatttg ctttactatt tatgtcatta tttttgccat tgtatcaaaa
      721 cctacacccg taatcacttt agttttactg gcaattattc ttggtctgcc cggcttaatt
      781 gttgttataa ctgctacgag atggtcgtac ctatggtgga tgtgcgtata tatttgtgct
      841 ttgcctattt ggaatttcgt actaccttca tatgcgtact ggaaatttga tgacttctca
      901 tggggtgata cgagaactat tgcgggaggt aataaaaagg cacaagacga gaatgaaggt
      961 gaatttgatc actcaaagat taaaatgagg acatggaggg aatttgaaag ggaagatatt
     1021 ctcaatcgga aggaggaaag tgactccttc gttgcatcgg atccaggaag ttcggggtcc
     1081 agcggttctt caggcgctag catggatagc actgagagcc tgttcaccgg ggtggtgccc
     1141 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     1201 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1261 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1321 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1381 caggagcgca ccatcacctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     1441 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1501 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacc
     1561 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa cgtggaggac
     1621 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1681 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     1741 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcagc
     1801 tccggctcca gcgggtctag catggatagc actgagagcc tgttcaccgg ggtggtgccc
     1861 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     1921 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1981 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2041 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2101 caggagcgca ccatcacctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     2161 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2221 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacc
     2281 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa cgtggaggac
     2341 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2401 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     2461 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcagc
     2521 tccggctcca gcgggtctag catggatagc actgagagcc tgttcaccgg ggtggtgccc
     2581 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     2641 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2701 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2761 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2821 caggagcgca ccatcacctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     2881 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2941 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacc
     3001 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa cgtggaggac
     3061 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3121 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     3181 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcagc
     3241 tccggctcca gcgggtaaag cggccgcaca gtatgaaaat atttttactg tgatacttac
     3301 aagttgatat atggttgtgt gtaacttatt tatttgagag gtattttaac acaccttaga
     3361 actaaaactt aataaataaa tatttctcta tctttaaagg cacatattac gtggctaagg
     3421 caattacagc tgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3481 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3541 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3601 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     3661 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3721 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3781 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3841 cgaaagggcg ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     3901 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     3961 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     4021 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     4081 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     4141 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     4201 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     4261 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     4321 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     4381 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     4441 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     4501 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     4561 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     4621 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     4681 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     4741 tcgtttcctg catgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     4801 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     4861 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     4921 agaataaaaa aaaaatgatg aattgaaaag ctggcctcgt gatacgccta tttttatagg
     4981 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5041 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5101 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5521 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5701 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6001 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6841 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6901 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
