LOCUS       sequence-200094-        9953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Vps16 with Vps16-iGFPx6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9953)
  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 9953)
  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-VPS16-iGFPx6
FEATURES             Location/Qualifiers
     source          1..9953
                     /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(6446..6463)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6446..6462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6455..6477)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6671..6690)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6790..6812
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6930..7733)
                     /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(7734..7954)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7961..7979)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8047..8151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8152..8220,8221..9012)
                     /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(8370..8389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9183..9771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9672..9691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9925..9942
                     /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 atgctggagg atcatgcacc caaagcaatt gaaatattaa agaattttgt cttagagaag
      301 ggtgtactgg actgtattgc tgctgcaata gacgaatttg aacctaaact acagaagatg
      361 ttgttgaacg ctgcatctta tggaaaagct tctttgcagt acaagtcatt tgacgctagt
      421 atatttgtga atgcgtgtaa tacgataaaa ttattaaact gcttcagaag ttttggtatt
      481 tttttgactg tggaggaata caggtgtatt tctttgaaag gtgtcattga taggctgttg
      541 aagtatcaca gatattatga gtgcatacag atatgcaaat tggccaatga acgatttctg
      601 ttaggatacg tgtttactga gtgggcaaag gataaaatca agggtagtcc tgacatggaa
      661 gatgatgagt tactagacaa aatcaaatct cgactaagtg ttatagatat gacggataca
      721 ttacaaatgg tagcagttgc gaaagttgca tatcttgaag gaaggttcca gttatcccgt
      781 aatttagctt tattagagaa aaacgaagaa gctagaattg agcaactgta taatcttgat
      841 gatgatagta tagcgctaaa ggagtgtatc aaagtacaaa attacagtct gacaatttca
      901 ttactgattg cactttccaa aaagctgaca aactctcaac tgacgaagct tttgatcatt
      961 gatatgttca acaatccctt atatctctat tatatgagaa tggataaagc atacttgtac
     1021 gatttttacc gccaaactga ccgatttatt gacttggctc acgtattact gcagcaggga
     1081 aaagaacagc aatctttaca cagcttttta ccgcaaatta aggaccttta tagccaagtt
     1141 caaaacagtg aagtggtcaa caatacaata gaacaacttc aaagacagga aaagttgtgg
     1201 atataccaag aatcattagg caaaagattt gctatatcat tcactaatat gactttagat
     1261 caaacgcttt cgaagttaat agaaacaggg caagataaac aggtgaaaga aatagttaaa
     1321 aagttcaaaa taagcgaaaa aaagctgtat cacctgaaat gtaaaacgtt agtcgaagct
     1381 aagaagtttg acgaactact acagtttgct cagtcgcgga aatcaccaat tggttacatg
     1441 cctttctaca catacttaaa aagcagaggc catatggata aagctagtcc ttatgttaat
     1501 atgatacctg gcctttcgta tcaggaaaag aaaaagcttt atgtggagtg caggggtttt
     1561 cgtgatgcaa ttcaactagc aggaaaggaa aaagacattc caggcttaaa ggagatatat
     1621 aatattatac cacctaatga gccggaatta aaggcattag caaatgaaac tatgagcagg
     1681 ataacggatc caggaagttc ggggtccagc ggttcttcag gcgctagcat ggatagcact
     1741 gagagccttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     1801 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     1861 aaatttattt gcactactgg aaaactacct gttccttggc caacacttgt cactacttta
     1921 acttatggtg ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc
     1981 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     2041 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     2101 ttaaaaggta ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac
     2161 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac
     2221 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     2281 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     2341 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta
     2401 acagctgctg ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact
     2461 gagagccttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     2521 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     2581 aaatttattt gcactactgg aaaactacct gttccttggc caacacttgt cactacttta
     2641 acttatggtg ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc
     2701 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     2761 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     2821 ttaaaaggta ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac
     2881 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac
     2941 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     3001 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     3061 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta
     3121 acagctgctg ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact
     3181 gagagccttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     3241 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     3301 aaatttattt gcactactgg aaaactacct gttccttggc caacacttgt cactacttta
     3361 acttatggtg ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc
     3421 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     3481 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     3541 ttaaaaggta ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac
     3601 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac
     3661 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     3721 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     3781 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta
     3841 acagctgctg ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact
     3901 gagagccttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     3961 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     4021 aaatttattt gcactactgg aaaactacct gttccttggc caacacttgt cactacttta
     4081 acttatggtg ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc
     4141 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     4201 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     4261 ttaaaaggta ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac
     4321 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac
     4381 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     4441 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     4501 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta
     4561 acagctgctg ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact
     4621 gagagccttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     4681 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     4741 aaatttattt gcactactgg aaaactacct gttccttggc caacacttgt cactacttta
     4801 acttatggtg ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc
     4861 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     4921 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     4981 ttaaaaggta ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac
     5041 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac
     5101 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     5161 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     5221 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta
     5281 acagctgctg ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact
     5341 gagagccttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     5401 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     5461 aaatttattt gcactactgg aaaactacct gttccttggc caacacttgt cactacttta
     5521 acttatggtg ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc
     5581 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     5641 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     5701 ttaaaaggta ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac
     5761 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac
     5821 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     5881 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     5941 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta
     6001 acagctgctg ggattacaca tggcagctcc ggctccagcg ggtaaagcgg ccgcaactca
     6061 taatttatag acgttgtgga tatctagccg catatcaaat aattaaatgc tgccattacg
     6121 tttgtgcggt tatatatgtt ggctccctat agggttaccc atggaacggc gaaggaaaaa
     6181 aaaaataatt ttcactgaaa aatttcaaga gtctatctca tctcatcgtt tattctgaat
     6241 taaatactac atacaaaatc tttttattag taaaatagtg agatttcttc gtgaactagt
     6301 gaaaagctat caatattacc aatctaccaa agtgagatgg aagaaacaat agaaaatgtt
     6361 gaagtgcctt ctagtaatgt ctcaaaacaa aatgacgacg gcctagatat gaagactttg
     6421 tttgtgaggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     6481 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     6541 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6601 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     6661 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     6721 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     6781 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     6841 gacgaaaggg cgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     6901 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     6961 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     7021 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     7081 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     7141 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     7201 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     7261 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     7321 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     7381 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     7441 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     7501 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     7561 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     7621 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     7681 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     7741 cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     7801 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     7861 tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa
     7921 aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg gcctcgtgat acgcctattt
     7981 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     8041 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     8101 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     8161 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     8221 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     8281 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8341 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8401 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8461 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8521 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8581 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8641 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8701 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8761 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8821 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8881 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8941 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     9001 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     9061 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     9121 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     9181 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     9241 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9301 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9361 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9421 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9481 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9541 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9601 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9661 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9721 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     9781 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     9841 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     9901 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aga
//
