LOCUS       sequence-202379-        5974 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 5974)
  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 5974)
  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..5974
                     /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(2472..2489)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2472..2488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2481..2503)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2697..2716)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2816..2838
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2956..3759)
                     /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
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCDQQN"
     promoter        complement(3760..3975)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3982..4000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4068..4172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4173..4241,4242..5033)
                     /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(4391..4410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5204..5792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5693..5712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5946..5963
                     /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 accgcaaata
     2281 ttattaaagc gtgttatttt ttttttctta ctcttctgta aacttctaaa tgctgctgag
     2341 gcacccctac tagcaatttc cctatcaaag tttgtttggc tgttattaag ggtatgtaag
     2401 acctcgtatc tgctgctact gatcaccatg ttagaaggag cagaatactt ttctctcgta
     2461 gttgggaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2521 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2581 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     2641 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2701 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2761 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2821 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcggg
     2881 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac
     2941 ttataataca gttttttagt tttgctggtc gcatcttctc aaatatgctt cccagcctgc
     3001 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc
     3061 aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc
     3121 caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc
     3181 ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct
     3241 cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga
     3301 caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt
     3361 caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc
     3421 tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa
     3481 ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac
     3541 tgtgccctcc atcgaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt
     3601 gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc
     3661 acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg
     3721 atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca
     3781 tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     3841 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     3901 ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa
     3961 aaatgatgaa ttgaaaagct ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4021 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4081 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4141 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4201 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4261 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4321 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4381 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4441 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4501 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4561 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4621 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4681 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4741 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4801 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4861 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4921 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4981 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5041 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5101 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5161 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5221 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5281 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5341 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5401 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5461 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5521 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5581 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5641 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5701 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5761 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5821 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5881 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5941 agcgcagcga gtcagtgagc gaggaagcgg aaga
//
