LOCUS       sequence-202562-        5151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses iGFP-PH(Osh1) under the TPI1 promoter. Integrating at 
            TRP1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5151)
  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 5151)
  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: YIplac204-T|C-iGFP-PH(OSH1)
FEATURES             Location/Qualifiers
     source          1..5151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        205..635
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(676..704)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      1716..1963
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1982..1998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1982..1998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1995..2017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2006..2022
                     /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)."
     promoter        complement(join(2030..2036,2037..2054,2055..2060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2075..2096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2213..2230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2384..2972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2464..2483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3143..3934,3935..4003))
                     /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     3766..3785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4004..4108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4176..4194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4196..4297
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4298..4972
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTSENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(5091..5113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
       61 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
      121 tgcgtaagga gaaaataccg catcaggcgg accttaatac attcagacac ttctgcggta
      181 tcaccctact tattcccttc gagattatat ctaggaaccc atcaggttgg tggaagatta
      241 cccgttctaa gacttttcag cttcctctat tgatgttaca cctggacacc ccttttctgg
      301 catccagttt ttaatcttca gtggcatgtg agattctccg aaattaatta aagcaatcac
      361 acaattctct cggataccac ctcggttgaa actgacaggt ggtttgttac gcatgctaat
      421 gcaaaggagc ctatatacct ttggctcggc tgctgtaaca gggaatataa agggcagcat
      481 aatttaggag tttagtgaac ttgcaacatt tactattttc ccttcttacg taaatatttt
      541 tctttttaat tctaaatcaa tctttttcaa ttttttgttt gtattctttt cttgcttaaa
      601 tctataacta caaaaaacac atacataaac taaaagaatt cgagcatgga tagcactgag
      661 agccttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
      721 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
      781 tttatttgca ctactggaaa actacctgtt ccttggccaa cacttgtcac tactttcact
      841 tatggtgttc agtgcttttc aagatacccg gatcatatga aacggcatga ctttttcaag
      901 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
      961 tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     1021 aaaggtattg attttaaaga agatggaaac attcttgggc acaaattgga atacaactat
     1081 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agctaacttc
     1141 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat
     1201 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     1261 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttaaaga gtttgtaaca
     1321 gctgctggga ttacacatgg cagctccggc tccagcgggt ccggaagtgg tagctcaggg
     1381 tctagtggat ccgaggcccc cacttacaaa ggctacctga aaaaatggac caacttcgct
     1441 caaggctaca aattgcgttg gttcatcctt agtagcgatg ggaaactatc ctactacatc
     1501 gatcaggccg acactaagaa tgcctgcagg ggctccctaa acatgtcttc gtgctctctg
     1561 catttggatt cgtctgaaaa gttgaaattc gaaattatcg gcggtaacaa cggtgttatc
     1621 aggtggcatt taaaggggaa ccaccccatc gagacaaata gatgggtttg ggccatccag
     1681 ggcgccataa gatacgcaaa ggacagatag agtcatcatg taattagtta tgtcacactt
     1741 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1801 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     1861 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     1921 cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcaagcttg gcgtaatcat
     1981 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2041 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2221 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2281 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2341 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2401 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2461 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2521 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2581 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2641 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2701 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2761 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2821 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2881 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2941 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3001 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3061 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3241 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3901 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3961 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4021 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4081 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4141 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccaattc
     4201 ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg
     4261 agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg
     4321 tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc
     4381 actagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat
     4441 tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg
     4501 cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct
     4561 ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata
     4621 ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat
     4681 actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc
     4741 aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga
     4801 gagccccgag agcttacatt ttatgttagc tggtggactg acgtcagaaa atgttggtga
     4861 tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg
     4921 tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac
     4981 tgagtagtat ttatttaagt attgtttgtg cacttgcctg caagcctttt gaaaagcaag
     5041 cataaaagat cgccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5101 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag c
//
