LOCUS       sequence-199903-        6998 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 6998)
  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 6998)
  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..6998
                     /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(3491..3508)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3491..3507)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3500..3522)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3716..3735)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3835..3857
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3975..4778)
                     /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(4779..4999)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5006..5024)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5092..5196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5197..5265,5266..6057)
                     /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(5415..5434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6228..6816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6717..6736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6970..6987
                     /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 acaacttgcc 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 actacattaa 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 tttttttttt accgttatta ttttgtttgt tagcatcttt
     3241 gattatctga ggggatttca aaatgccaat aaaaaagacc ttatgagttc tgtatttttc
     3301 ttaatatttt cgagagaatt tcccaactat aaaattgtag attacaggaa taaataaatt
     3361 gcagaaacta aaatgattga aactgccttt tcaatttata tacttttata tcaaattatg
     3421 ttcaactcta gtagagtttc ttttattacg caaacgtcct atgaccttga ccggtaccga
     3481 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3541 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3601 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     3661 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     3721 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3781 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3841 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcgggt
     3901 aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact
     3961 tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct
     4021 tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca
     4081 acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc
     4141 aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct
     4201 tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc
     4261 ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac
     4321 aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc
     4381 aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct
     4441 gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat
     4501 tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact
     4561 gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg
     4621 ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca
     4681 cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga
     4741 tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcat
     4801 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
     4861 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     4921 gagattaccg aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa
     4981 aatgatgaat tgaattgaaa agctggcctc gtgatacgcc tatttttata ggttaatgtc
     5041 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5701 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6061 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6121 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6361 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6841 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6901 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6961 accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
