LOCUS       sequence-202382-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Ypt31 with iGFP-Ypt31..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  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 5636)
  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-iGFP-YPT31
FEATURES             Location/Qualifiers
     source          1..5636
                     /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(578..606)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(2134..2151)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2134..2150)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2143..2165)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2359..2378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2478..2500
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2618..3421)
                     /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(3422..3637)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3644..3662)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3730..3834
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3835..3903,3904..4695)
                     /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(4053..4072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4866..5454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5355..5374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5608..5625
                     /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 atgcttcaga tctatcccag cgcaatctaa ttgcactaat taggtagtag tcttgctgta
      301 agtatgttgt attgttgttc ctaggcgtca attctactgt tgttatgttt gggctcgttt
      361 gttgtttgtt taccattttc caatgcggaa tatcacaggc cgctgcggtg tgcgggtgct
      421 aaattagaga aagagggaaa ctcaaaactt ttcaagaaag aaacgaataa caatttgacc
      481 tttattacaa ggcactttgt ttaggccagc aaagggattc tgacggcgtc tggggatttc
      541 aacaatgagc gatagcactg agagcctttt cactggagtt gtcccaattc ttgttgaatt
      601 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      661 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccttggcc
      721 aacacttgtc actactttca cttatggtgt tcagtgcttt tcaagatacc cggatcatat
      781 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
      841 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
      901 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
      961 gcacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     1021 gaatggaatc aaagctaact tcaaaattag acacaacatt gaagatggaa gcgttcaact
     1081 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     1141 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     1201 ggtccttaaa gagtttgtaa cagctgctgg gattacacat ggcagctccg gctccagcgg
     1261 gtccggaagt ggtagctcag ggtctagtgg aagcgaggac tacgggtatg attacgacct
     1321 actatttaaa attgtactga tcggtgattc cggggtaggg aaatccaatc tattgtcaag
     1381 atttacgaag aacgaattca atatggattc aaagtctaca attggggttg agtttgccac
     1441 cagaactttg gaaattgatg gcaagaggat caaggcgcag atttgggata ctgcgggcca
     1501 ggaacgttac agagccatta cttctgctta ttatagagga gctgtaggtg ccttgatcgt
     1561 ttacgatatc agtaaatcaa gctcatatga aaattgtaat cattggttat cagaattaag
     1621 agaaaatgca gatgacaacg tcgcagtggg gcttatcggt aacaagtctg atttggccca
     1681 tttacgtgct gtacctacgg aggagtccaa gacttttgca caggaaaatc agttattatt
     1741 cacagaaaca agtgctttga acagtgaaaa cgtggataag gcttttgaag agttaatcaa
     1801 taccatctat caaaaggtca gcaagcatca aatggacctt ggtgactcca gtgccaacgg
     1861 gaatgccaat ggtgcgtccg ctcctaatgg accaacgatt agtctgacac caacaccaaa
     1921 cgaaaacaaa aaggcaaatg gaaataactg ttgttaagag atattttgca gcagttgcgc
     1981 acttgcatgt gaatgactct tctccccttt aattctgtgc tatattttta caattttctg
     2041 ctgacatata gtttatatac atatagaacg catataggaa attgaagtaa acagaataca
     2101 caagtagagg ccggtatgta cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga
     2161 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2221 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2281 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2341 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2401 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2461 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     2521 acgcgcgaga cgaaagggcg ggtaataact gatataatta aattgaagct ctaatttgtg
     2581 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     2641 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     2701 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     2761 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     2821 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     2881 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     2941 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     3001 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     3061 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     3121 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     3181 ataatgcctt tagcggctta actgtgccct ccatcgaaaa atcagtcaag atatccacat
     3241 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     3301 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     3361 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     3421 tgatttatct tcgtttcctg catgtttttg ttctgtgcag ttgggttaag aatactgggc
     3481 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     3541 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     3601 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctggcctcgt gatacgccta
     3661 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3721 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3781 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3841 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3901 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3961 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4021 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4081 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4141 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4201 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4261 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4321 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4381 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4441 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4501 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4561 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4621 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4681 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4741 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4801 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4981 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5341 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5401 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5461 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5521 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5581 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
