LOCUS       sequence-202394-        5991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Vps21 with msGFP-Vps21..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  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 5991)
  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-msGFP-VPS21
FEATURES             Location/Qualifiers
     source          1..5991
                     /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(618..639)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1226..1247
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2489..2506)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2489..2505)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2498..2520)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2714..2733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2833..2855
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2973..3776)
                     /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
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRGCGQQN"
     promoter        complement(3777..3992)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3999..4017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4085..4189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4190..4258,4259..5050)
                     /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(4408..4427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5221..5809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5710..5729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5963..5980
                     /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 atgccggagg caacaacaaa gccaaaacct agattatgac gatataaact tatgctgagg
      301 taggtatata ccagaggatt tattttatag atattagtct gctgattatc tagtttcttg
      361 tcacctcata cgtgtttttt tttttttggt tttctcggtt tgcattatgg ggctttttgc
      421 taaaacaata tccccgaaaa gacaaagaaa gagaagtgag gtgaaaatac gagcagctta
      481 tcttataaaa gtggcaacga ttttaaaaat aaaaagcaag atcgtcgact tatctaagct
      541 ataaaaaaat atccctttta tcacacaaaa aaatgaacga tagcactgag agcctgttca
      601 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      661 tgcgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca
      721 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      781 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      841 ccgaaggcta cgtccaggag cgcaccatca ccttcaagga cgacggcacc tacaagaccc
      901 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      961 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca
     1021 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1081 acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1141 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     1201 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1261 tcactcacgg cagctccggc tccagcgggt ccggaagtgg tagctcaggg tctagtggat
     1321 ccacatcagt cacttccata aagttggtac tgttgggtga ggcagcagtt ggtaaatcgt
     1381 caatagtcct aaggttcgta tctaatgatt ttgccgaaaa taaggagccc accattggtg
     1441 cagcattcct cactcaaaga gttactataa atgaacatac tgttaagttt gaaatatggg
     1501 acactgctgg gcaagagaga tttgcatctt tagcacctat gtactataga aacgcacaag
     1561 ctgctcttgt agtatacgac gttacaaaac cacaatcctt tattaaagcg cgccactggg
     1621 ttaaggaatt acatgaacag gcatcaaagg atatcatcat tgcgttagtt ggtaacaaga
     1681 tcgatatgct gcaagaaggt ggggaaagaa aagtcgcaag ggaagaaggt gaaaagctag
     1741 cggaagaaaa gggcttgttg ttctttgaaa caagtgcaaa gacaggagag aatgttaacg
     1801 atgtattttt gggaatcggt gaaaagatac cactgaagac ggctgaagag caaaatagtg
     1861 caagtaacga acgtgagtcg aacaatcaaa gagtggattt gaacgctgct aatgatggga
     1921 ccagcgcaaa cagtgcttgc agttgttaga gaggggaaaa taaatatata taaaaaaaaa
     1981 gtctgggcaa attgagataa atagatcttc aaaacgggaa agatgaaagt cataatttat
     2041 cgtacctaga aaggagaggg agttttcatt ttttaccgta atcgttgttt ttcttttctt
     2101 cttataattt gtaaattttt taactgcatt agatcattat attatctttc aaaacgccag
     2161 tcgtttttat tagaccttta tatacgtttc aaagtggttt gtacacgtta gtttagttaa
     2221 acttttgttt tcatgtagac caaaatataa acttatttta tgccgaaaat cgggagatgg
     2281 attagttaat attgtaaata atgataaatt aaaggaacat taaatttaca aagttgcaat
     2341 gaggagcaat atatatatat atatatggat ttctgagaaa ttaattaagc agtatttgaa
     2401 cacatgtcgt ttactctttc ttatccttta tgtctaattt ttcgagtaga tcggtaatta
     2461 cagctgtcaa ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
     2521 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2581 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2641 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     2701 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2761 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2821 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2881 cgagacgaaa gggcgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     2941 agtatacatg catttactta taatacagtc ttttagtttt gctggccgca tcctctcaaa
     3001 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     3061 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     3121 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     3181 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     3241 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     3301 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     3361 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     3421 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     3481 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     3541 gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt
     3601 tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta
     3661 cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg
     3721 gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt
     3781 tatcttcgtt tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     3841 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     3901 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa
     3961 aaaaaagaat aaaaaaaaaa tgatgaattg aaaagctggc ctcgtgatac gcctattttt
     4021 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4741 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4801 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5041 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5881 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5941 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a
//
