LOCUS       sequence-202380-        6492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Tlg2 with iGFP-Tlg2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  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 6492)
  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-iGFP-TLG2
FEATURES             Location/Qualifiers
     source          1..6492
                     /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(621..649)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(2990..3007)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2990..3006)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2999..3021)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3215..3234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3334..3356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3474..4277)
                     /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(4278..4493)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4500..4518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4586..4690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4691..4759,4760..5551)
                     /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(4909..4928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5722..6310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6211..6230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6464..6481
                     /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 ccaagcttaa
      241 gagttagtcg ttgatatggt attctctaat accgaaaata acttagtaca tataaacgcg
      301 taaagttcac gcgtttatat caccctacca tatgacattc taaccctact aattttttat
      361 caaagctaat agggttgtat tttattatct tttacaccta cctttttttt ttgtatatgc
      421 cctatagatg aattataatt gttggaccgg tttgtattgg gcttttaagg tgtaaagtga
      481 gtgcgtatag attatagcag aatagactat aagtacggca agacaggaaa gtctccaatt
      541 gatgtacatt ttatcgcttt gtgatctagg cagtcgttac aaacatgttt atggatagca
      601 ctgagagcct tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
      661 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc
      721 ttaaatttat ttgcactact ggaaaactac ctgttccttg gccaacactt gtcactactt
      781 tcacttatgg tgttcagtgc ttttcaagat acccggatca tatgaaacgg catgactttt
      841 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
      901 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
      961 agttaaaagg tattgatttt aaagaagatg gaaacattct tgggcacaaa ttggaataca
     1021 actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagcta
     1081 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
     1141 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
     1201 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt aaagagtttg
     1261 taacagctgc tgggattaca catggcagct ccggctccag cgggtccgga agtggtagct
     1321 cagggtctag tggatccaga gatagaacta atttattttt atcataccgt aggactttcc
     1381 ctcataacat aactttctca tctgggaaag cacccttggg cgatgatcaa gatattgaaa
     1441 tgggtacata cccgatgatg aatatgtctc acgatatatc ggccagattg acagatgaac
     1501 gaaaaaacaa gcatgagaat cacagtgatg cgttaccacc catatttatt gatatagcgc
     1561 aggatgttga tgattatctt ctggaggtga gacggttaag tgagcagttg gcgaaggttt
     1621 ataggaagaa ttcgttacct ggctttgaag acaagagtca tgatgaggcg ctgattgaag
     1681 atctaagttt caaagtaatt cagatgctgc agaaatgtta cgctgttatg aaacgattaa
     1741 agaccatata caactctcaa tttgtggatg gtaagcaatt aagtcgagaa gagctgatta
     1801 tactagataa tctacagaag atctatgcag aaaagataca aactgaaagt aataagttca
     1861 gagtattaca aaataattac ttgaaattct tgaacaagga tgatttgaaa cctatccgca
     1921 ataaggccag tgcagagaac acgctattat tagatgatga ggaagaggag gcagcaagag
     1981 agaagagaga gggcctcgat attgaagact attcaaaaag gacgttgcag agacagcaac
     2041 agcttcatga cacttctgca gaagcatatt tgagagaacg tgacgaagaa attactcagt
     2101 tggcaagggg tgtattagaa gtgagcacta ttttcaggga aatgcaagat ttggttgttg
     2161 accaggggac tattgtggac aggattgatt ataatctgga aaatacagtg gtagagctaa
     2221 aaagtgcaga taaagagttg aataaagcaa cacactacca gaagagaact caaaaatgta
     2281 aagtcatttt attactaaca ttgtgcgtaa tagcgctctt tttctttgtt atgttgaaac
     2341 cacatggcgg tggcagtggt ggtcgtaata atggtagcaa caagtacaac aatgatgata
     2401 ataaaacagt taataacagt catgatgacg gcagtaatac ccatattaat gacgaagaga
     2461 gcaacttgcc gagtatcgtt gaagtaacag aatctgagaa tgatgctttg gatgacctac
     2521 tttgatgaca aaactttcac ggtaccgcat cgcgattttg aactataata tgctttgaag
     2581 aaaattgaga gatttagtca gcgttcatat gacctgttgc tcgttctatt tgggaaaagt
     2641 gaaaaggtta atatatgatt tcaataaagt atgtagtatg ataataaaat acaaatttat
     2701 atgggataaa attgaaccgc aagtaaagac tggcgaacga atattgtgca taagtagttc
     2761 ctgaagatga taaaatatgg tttaatacaa gttggtggtt attgtgattt gtagaatata
     2821 aaaaaaaaaa aagaaccttc atttttcttt ttttacctaa atccgtaagg attgttcagt
     2881 gaagtagtag ctcggggcaa gtttttgttt tgcattctcc tcttataggc aggcttatat
     2941 ccttgtgtta tcgggctttg gtttggtaaa ggattcggag ctcgaattca ctggccgtcg
     3001 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3061 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3121 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     3181 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3241 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3301 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3361 caccgtcatc accgaaacgc gcgagacgaa agggcgggta ataactgata taattaaatt
     3421 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     3481 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     3541 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     3601 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     3661 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     3721 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     3781 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     3841 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     3901 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     3961 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     4021 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat cgaaaaatca
     4081 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     4141 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     4201 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     4261 tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct gtgcagttgg
     4321 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     4381 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     4441 tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaagctgg
     4501 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4561 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4621 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4681 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4741 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4801 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4861 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4921 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4981 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5041 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5101 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5161 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5221 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5281 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5341 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5401 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5461 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5521 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5581 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5641 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5701 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5761 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5821 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5881 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5941 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6001 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6061 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6121 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6181 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6241 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6301 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6361 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6421 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6481 ggaagcggaa ga
//
