LOCUS       sequence-202546-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Chc1 with Chc1-msGFPx3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  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 6992)
  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-CHC1-msGFPx3
FEATURES             Location/Qualifiers
     source          1..6992
                     /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(3490..3507)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3490..3506)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3499..3521)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3715..3734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3834..3856
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3974..4777)
                     /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(4778..4993)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5000..5018)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5086..5190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5191..5259,5260..6051)
                     /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(5409..5428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6222..6810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6711..6730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6964..6981
                     /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 atgcctgcag gtcgactcta gttgatcttt tgacctcatt gacaccaaga ttagatattc
      301 caagaactgt caaaattttc tcgaaatccg acaactcgcc tttaatcaag cctttcttga
      361 tcaatgtctt accaaaaaat aactcagttg tcaaccaagc ctaccacgat ctgatgattg
      421 aagaagaaga ttataaagct ttgcaagacg ctgttgattc ttatgacaaa tttgaccaat
      481 tgggattggc atctcgtttg gaatctcata agctaatttt tttcaagaaa ataggagccc
      541 tgttataccg tagaaataag aagtgggcaa agagtctttc aatcttgaag gaagaaaagc
      601 tatggaagga cgctattgag acggcagcaa tctctcagga tccaaaggta gtggaggctc
      661 tactcacata tttcgttgaa accggtaaca gagaaggctt tgttgctttg ctatatgctg
      721 cttacaactt agtgagaatt gaatttgttt tagaaatatc atggatgaat tctttggagg
      781 actgcattaa gccatttgaa atctccataa aaaaagagca gaatgattcc attaagaaaa
      841 tcaccgaaga actcgctaaa aagtccggat ctaatgaaga gcacaaagat ggccaacctt
      901 tgatgctgat gaacagcgcg atgaacgttc aacccacagg atttacagat ccaggaagtt
      961 cggggtccag cggttcttca ggcgctagca tggatagcac tgagagcctg ttcaccgggg
     1021 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg
     1081 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     1141 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1201 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1261 gctacgtcca ggagcgcacc atcaccttca aggacgacgg cacctacaag acccgcgccg
     1321 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1381 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacgtct
     1441 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaacg
     1501 tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1561 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     1621 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1681 acggcagctc cggctccagc gggtctagca tggatagcac tgagagcctg ttcaccgggg
     1741 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg
     1801 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     1861 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1921 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1981 gctacgtcca ggagcgcacc atcaccttca aggacgacgg cacctacaag acccgcgccg
     2041 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2101 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacgtct
     2161 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaacg
     2221 tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2281 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     2341 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2401 acggcagctc cggctccagc gggtctagca tggatagcac tgagagcctg ttcaccgggg
     2461 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg
     2521 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     2581 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2641 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2701 gctacgtcca ggagcgcacc atcaccttca aggacgacgg cacctacaag acccgcgccg
     2761 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2821 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacgtct
     2881 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaacg
     2941 tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3001 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     3061 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3121 acggcagctc cggctccagc gggtaaagcg gccgcgacgt ggataaaata attcgtttgc
     3181 tgtaccccat cgtgtatttt ttttttttta ccgttattat tttgtttgtt agcatctttg
     3241 attatctgag gggatttcaa aatgccaata aaaaagacct tatgagttct gtatttttct
     3301 taatattttc gagagaattt cccaactata aaattgtaga ttacaggaat aaataaattg
     3361 cagaaactaa aatgattgaa actgcctttt caatttatat acttttatat caaattatgt
     3421 tcaactctag tagagtttct tttattacgc aaacgtccta tgaccttgac cggtaccgag
     3481 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3541 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3601 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     3661 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     3721 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3781 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3841 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcgggta
     3901 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
     3961 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
     4021 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
     4081 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
     4141 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
     4201 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
     4261 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
     4321 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
     4381 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     4441 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     4501 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     4561 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     4621 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     4681 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     4741 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg
     4801 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     4861 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     4921 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
     4981 atgatgaatt gaaaagctgg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5041 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5101 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5161 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5221 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5281 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5341 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5401 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5461 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5521 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5581 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5641 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5701 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5761 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5821 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5881 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5941 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6001 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6061 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6121 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6901 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6961 cgcagcgagt cagtgagcga ggaagcggaa ga
//
