LOCUS       sequence-199929-        5978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Tlg1 with iGFP-Tlg1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  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 5978)
  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-TLG1
FEATURES             Location/Qualifiers
     source          1..5978
                     /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(700..728)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(2471..2488)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2471..2487)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2480..2502)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2696..2715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2815..2837
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2955..3758)
                     /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(3759..3979)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3986..4004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4072..4176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4177..4245,4246..5037)
                     /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(4395..4414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5208..5796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5697..5716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5950..5967
                     /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 atgcttcgcg ttgaatttgc tccattggaa tatcagcatt acttgaagta tcctctacca
      301 taggaaccgt aacttcatta tgttggggac tattctcact ttcattcatt gtataggttt
      361 ccctaattct cttaagagaa cctgcttatg aaggcttttt tttgtttgga tcaccatctg
      421 tttttctttt tttatcgagt ttttaattat ccacgagata ttcgaaccta cgacataaag
      481 tagtaaatat cagaagatag gacaattcga agcacacaac acagaacata tactaaaatt
      541 atatttgctg tctttctgac aaggtttgtg aacattttaa tttctacgtt ttccctgtaa
      601 ctgaagtaaa cgtgaaagac gtttaactgt aaaattgtat tgctaagatc ctttctttaa
      661 caaatgaaca tggatagcac tgagagcctt ttcactggag ttgtcccaat tcttgttgaa
      721 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      781 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccttgg
      841 ccaacacttg tcactacttt cacttatggt gttcagtgct tttcaagata cccggatcat
      901 atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
      961 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     1021 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     1081 gggcacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     1141 aagaatggaa tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     1201 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1261 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     1321 atggtcctta aagagtttgt aacagctgct gggattacac atggcagctc cggctccagc
     1381 gggtccggaa gtggtagctc agggtctagt ggatccaaca gtgaagatcc gtttcaacaa
     1441 gttgttaagg acaccaagga gcaattgaac cgcataaaca attacataac tcgtcacaat
     1501 actgctggtg atgacgatca agaggaggaa atacaagata ttttaaagga tgttgaggaa
     1561 acaatagttg atttggacag aagcataatc gtaatgaaaa gggatgaaaa cgaagacgtg
     1621 agtggtaggg aagcacaagt taaaaatata aaacagcaac ttgatgctct gaagttgcgt
     1681 tttgatcgaa gaatacagga atctactcaa acaactattc ctctagaaga gacggtggaa
     1741 aattcaacac ttaacaccag catggctgag aacaatgatg gtggtatgtc caatccgttt
     1801 caggaacaaa tgttaagaga acaagatgtt catttagatg gtattcacaa gacaatgcaa
     1861 aatttgcata ttcaagctca aacaatgggg gatgaattag agaaccaggg acaattgttg
     1921 gataatatgg acgagggtat ggacggtgtt gtaaataagc tggctagagg tcgtaggcaa
     1981 ttggaatggg tctacgaaaa aaataaagaa aaatacgacg attgttgtat aggacttctt
     2041 attgtcgtct tgatagtttt attagttttg gcattcattg cttgaaaaat acaatgcact
     2101 aatgccgaag agcttcaatc gaagatatac tgggcatata taaaatacat gcattctgaa
     2161 tacataagtg atacggatat taatgctaca agggtttgga tatgatttat atgttggttt
     2221 ttctggatcg tcaacaactc gtgcatattt ttttgtttcg aagtagagga accacaaata
     2281 ttattaaagc gtgttatttt tttttcttac tcttctgtaa acttctaaat gctgctgagg
     2341 cacccctact agcaatttcc ctatcaaagt ttgtttggct gttattaagg gtatgtaaga
     2401 cctcgtatct gctgctactg atcaccatgt tagaaggagc agaatacttt tctctcgtag
     2461 ttgggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2521 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2581 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     2641 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     2701 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2761 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2821 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcgggt
     2881 aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact
     2941 tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct
     3001 tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca
     3061 acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc
     3121 aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct
     3181 tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc
     3241 ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac
     3301 aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc
     3361 aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct
     3421 gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat
     3481 tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact
     3541 gtgccctcca ttgaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg
     3601 ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca
     3661 cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga
     3721 tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcat
     3781 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
     3841 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     3901 gagattaccg aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa
     3961 aatgatgaat tgaattgaaa agctggcctc gtgatacgcc tatttttata ggttaatgtc
     4021 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4141 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4201 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4261 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4321 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4381 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4441 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4501 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4561 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4621 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4681 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4741 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4801 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4861 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4921 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4981 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5041 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5101 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5161 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5221 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5281 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5341 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5401 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5461 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5521 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5581 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5641 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5701 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5761 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5821 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5881 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5941 accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
