LOCUS       sequence-200091-        9202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Vps8 with Vps8-iYFPx6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9202)
  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 9202)
  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-VPS8-iYFPx6
FEATURES             Location/Qualifiers
     source          1..9202
                     /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(5695..5712)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5695..5711)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5704..5726)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5920..5939)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6039..6061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6179..6982)
                     /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(6983..7203)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7210..7228)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7296..7400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7401..7469,7470..8261)
                     /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(7619..7638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8432..9020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8921..8940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9174..9191
                     /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 atgcagttct cagcaacgct gagagcttac aaattctgga cttattgaat caagactcta
      301 attttgaagc tgcagctata attcacgaac gcttggaaag ttttaaccta gcagtcaggg
      361 atttattaag ttttattgaa caatgtctaa atgaagggaa aacaaatata tctactttat
      421 tggaatcttt gaggagggcc tttgatgatt gtaattctgc tggtaccgag aaaaaatcgt
      481 gttggatatt attgattaca ttcctgatca ctctatatgg gaaatatcct tcacacgatg
      541 aaaggaaaga tttatgtaat aaactacttc aagaagcatt tttgggattg gttaggtcca
      601 agagttcctc tcagaaggat tcaggtgggg aattctggga aataatgtct tctgttcttg
      661 agcaccaaga cgttatttta atgaaagttc aggatttaaa gcaactgcta ctgaatgttt
      721 ttaatactta taaattggaa agatctcttt ctgagttgat tcaaaagatt atagaggatt
      781 cttcgcaaga tcttgttcaa cagtatagaa aatttctgag tgaagggtgg tctatacaca
      841 ccgacgactg cgaaatctgc gggaaaaaaa tatggggagc tggtctggac ccattacttt
      901 ttctagcttg ggaaaatgta cagcgccacc aagatatgat tagtgtagat ctcaaaactc
      961 cccttgtcat attcaaatgt caccatggct ttcaccagac ttgcctcgaa aacttggccc
     1021 agaaacccga tgaatattct tgtttaattt gccagacgga atctaaccca aaaatagtaa
     1081 cggatccagg aagttcgggg tccagcggtt cttcaggcgc tagcatggat agcactgaga
     1141 gccttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     1201 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     1261 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaggtt
     1321 atggtttgca gtgctttgca agatacccgg atcatatgaa acagcatgac tttttcaaga
     1381 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     1441 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     1501 aaggtattga ttttaaagaa gatggaaaca ttcttgggca caaattggaa tacaactata
     1561 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     1621 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata
     1681 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
     1741 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttaaagag tttgtaacag
     1801 ctgctgggat tacacatggc agctccggct ccagcgggtc tagcatggat agcactgaga
     1861 gccttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     1921 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     1981 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaggtt
     2041 atggtttgca gtgctttgca agatacccgg atcatatgaa acagcatgac tttttcaaga
     2101 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     2161 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     2221 aaggtattga ttttaaagaa gatggaaaca ttcttgggca caaattggaa tacaactata
     2281 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     2341 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata
     2401 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
     2461 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttaaagag tttgtaacag
     2521 ctgctgggat tacacatggc agctccggct ccagcgggtc tagcatggat agcactgaga
     2581 gccttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     2641 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     2701 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaggtt
     2761 atggtttgca gtgctttgca agatacccgg atcatatgaa acagcatgac tttttcaaga
     2821 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     2881 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     2941 aaggtattga ttttaaagaa gatggaaaca ttcttgggca caaattggaa tacaactata
     3001 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     3061 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata
     3121 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
     3181 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttaaagag tttgtaacag
     3241 ctgctgggat tacacatggc agctccggct ccagcgggtc tagcatggat agcactgaga
     3301 gccttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     3361 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     3421 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaggtt
     3481 atggtttgca gtgctttgca agatacccgg atcatatgaa acagcatgac tttttcaaga
     3541 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     3601 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     3661 aaggtattga ttttaaagaa gatggaaaca ttcttgggca caaattggaa tacaactata
     3721 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     3781 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata
     3841 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
     3901 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttaaagag tttgtaacag
     3961 ctgctgggat tacacatggc agctccggct ccagcgggtc tagcatggat agcactgaga
     4021 gccttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     4081 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     4141 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaggtt
     4201 atggtttgca gtgctttgca agatacccgg atcatatgaa acagcatgac tttttcaaga
     4261 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     4321 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     4381 aaggtattga ttttaaagaa gatggaaaca ttcttgggca caaattggaa tacaactata
     4441 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     4501 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata
     4561 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
     4621 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttaaagag tttgtaacag
     4681 ctgctgggat tacacatggc agctccggct ccagcgggtc tagcatggat agcactgaga
     4741 gccttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     4801 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     4861 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaggtt
     4921 atggtttgca gtgctttgca agatacccgg atcatatgaa acagcatgac tttttcaaga
     4981 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     5041 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     5101 aaggtattga ttttaaagaa gatggaaaca ttcttgggca caaattggaa tacaactata
     5161 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     5221 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata
     5281 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
     5341 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttaaagag tttgtaacag
     5401 ctgctgggat tacacatggc agctccggct ccagcgggta aagcggccgc catttctaaa
     5461 tatttaatac aactttggtt acataaaagt aaaatttata cacctcattt cattatgtag
     5521 attcatatat agaataccaa ttatgattga cccaatagcc atcaaaatca gtagttatta
     5581 atacttgtct ttctaggagc catttgcata tttctgatat ttcatgaagc gaaagtactt
     5641 cacgacacct agattgcaat ctactcaatg ttatccctgg atgagctcga attcactggc
     5701 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5761 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5821 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     5881 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     5941 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6061 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc gggtaataac tgatataatt
     6121 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
     6181 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
     6241 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
     6301 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     6361 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     6421 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     6481 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
     6541 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
     6601 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     6661 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     6721 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     6781 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     6841 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     6901 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     6961 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcatgttttt gttctgtgca
     7021 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     7081 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     7141 aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt
     7201 gaaaagctgg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     7261 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7321 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     7381 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     7441 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     7501 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7561 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     7621 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     7681 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     7741 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     7801 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     7861 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7921 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7981 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     8041 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     8101 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     8161 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     8221 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     8281 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     8341 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8401 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8461 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8521 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     8581 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8641 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8701 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8761 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8821 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8881 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8941 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9001 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9061 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     9121 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     9181 cagtgagcga ggaagcggaa ga
//
