LOCUS       sequence-202383-        5446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for integration of iGFP-Ypt52 at TRP1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5446)
  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 5446)
  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-iGFP-YPT52
FEATURES             Location/Qualifiers
     source          1..5446
                     /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(714..742)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(2191..2208)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2191..2207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2200..2222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2416..2435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2535..2557
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2676..3350)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3351..3452)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3454..3472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3540..3644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3645..3713,3714..4505)
                     /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(3863..3882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4676..5264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5165..5184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5418..5435
                     /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 atgcgttccc atgttcaaat gtgtgaaagg taggtaaaat cattatgtta aattttgaaa
      301 tccacatgac cttgaattaa ataggcacat cacgattgta aaggaatgaa atcaatctga
      361 ctccaaatta ctcaatgttc ttaacactat gtgtatcatt gagaccaaac ggcaacttca
      421 tagtgtcttg ctttcgtgga cagcgactac tattctggcc ggacaattca ggtttataaa
      481 gctctgcccc tgcgatatga agttacaatg ttaacatacg ggagtgaagg gaacaggccc
      541 gtttaccaac tgtcagtatt aattatttgt taagtcgctt cgatatcgag ctctggacat
      601 atctaaattt ttttttatag tgcgagggta ggaagaaggt ttaacttatt tggagtaaac
      661 gtatatatta ttaacagatg ttgatggata gcactgagag ccttttcact ggagttgtcc
      721 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
      781 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
      841 tacctgttcc ttggccaaca cttgtcacta ctttcactta tggtgttcag tgcttttcaa
      901 gatacccgga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
      961 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
     1021 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
     1081 atggaaacat tcttgggcac aaattggaat acaactataa ctcacacaat gtatacatca
     1141 tggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattagacac aacattgaag
     1201 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     1261 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     1321 aaaagagaga ccacatggtc cttaaagagt ttgtaacagc tgctgggatt acacatggca
     1381 gctccggctc cagcgggtcc ggaagtggta gctcagggtc tagtggatcc caatttaaac
     1441 tagttttatt gggtgactca tccgtgggta aatcttccat tgttcatcgt tttgttaagg
     1501 atacattcga tgaactacgt gaaagtacga ttggtgctgc ttttttatct caatcaatca
     1561 ctattcaccc aaacgatggt aatgaaacaa aggatgtcgt tataaaattt gaaatctggg
     1621 atactgctgg tcaagaaaga tacaagtcat tggcaccaat gtattataga aacgctaatg
     1681 cagcattagt tgtttatgac attacgcagg aagattcact acaaaaggca aggaactggg
     1741 ttgatgagct gaaaaataag gtaggcgacg acgacttggt tatttattta ctgggaaaca
     1801 aagtggattt atgtcaagaa acgcctagca ctgagacaag cccagattca aacgaaggtg
     1861 gagatgaaga acaaaaagta agagcaatta gcactgaaga agcgaaacag tacgcacaag
     1921 aacaagggtt gctgttccga gaagttagtg ctaagacagg tgaaggggtt aaggaaattt
     1981 ttcaagatat tggtgaaaaa ttatacgatt taaagaaaga cgaaattctc tctaagcaaa
     2041 atagacaaat cggtggaggc aataacggac aagtagacat taatttgcaa aggccatcca
     2101 caaatgatcc aacatcttgt tgcagttgat atcctatgat ttgtgtttta gaggaaaaac
     2161 aacagaagtt gcggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     2221 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2281 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     2341 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2401 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     2461 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2521 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2581 cgcgagacga aagggcgatc ttttatgctt gcttttcaaa aggcttgcag gcaagtgcac
     2641 aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa
     2701 tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca
     2761 ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct
     2821 aacataaaat gtaagctctc ggggctctct tgccttccaa cccagtcaga aatcgagttc
     2881 caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga
     2941 ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta
     3001 ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg
     3061 acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga
     3121 aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt
     3181 gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata
     3241 cccagcaagt cagcatcgga atctagtgca cattctgcgg cctctgtgct ctgcaagccg
     3301 caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct
     3361 gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct
     3421 tggccctctc cttttctttt ttcgaccgaa ttggcctcgt gatacgccta tttttatagg
     3481 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3541 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3601 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3661 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3721 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3781 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3841 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3901 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3961 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4021 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4081 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4141 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4201 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4261 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4321 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4381 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4441 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4501 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4561 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4621 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4681 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4741 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4801 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4861 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4921 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4981 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5041 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5101 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5161 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5221 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5281 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5341 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5401 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
