LOCUS       sequence-202456-        9826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Hse1 with Hse1-iGFPx6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9826)
  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 9826)
  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-HSE1-iGFPx6
FEATURES             Location/Qualifiers
     source          1..9826
                     /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(6324..6341)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6324..6340)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6333..6355)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6549..6568)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6668..6690
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6808..7611)
                     /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(7612..7827)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7834..7852)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7920..8024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8025..8093,8094..8885)
                     /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(8243..8262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9056..9644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9545..9564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9798..9815
                     /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 atgctaacgt gatcttaaga accctctctt tgacggtatc tttggcagaa aattgtggtt
      301 cccgcttaag acaggaaatt agttcaaaaa attttacctc tttgttatat gcgctcatcg
      361 agagccattc tgtacacatt actctaaaaa aagcggtgac tgatgttgtc aagcaactgt
      421 ctgattcatt caaggatgac ccatcattac gcgccatggg agatctatat gacaaaatta
      481 aaagaaaggc tccatatttg gtacagccta atgtaccaga gaagcataac atgagtaccc
      541 aggcagacaa ctcagatgat gaggaattac aaaaagcact aaaaatgtct ttgtttgagt
      601 acgaaaagca aaaaaagcta caagaacagg aaaaagaaag tgccgaagtc cttccgcagc
      661 aacagcagca gcatcagcag cagaatcaag ctcctgctca caaaataccg gcacaaaccg
      721 ttgtgagaag agttcgggct ctatatgact taaccacaaa cgaaccggat gaattgtcct
      781 ttaggaaggg cgatgttatt acagtgctgg aacaagtata tagagattgg tggaagggag
      841 ctcttcgagg aaatatgggt atatttccat taaattatgt aacaccgatt gttgagccat
      901 cgaaagaaga aattgagaag gaaaaaaata aggaagccat agtattctct caaaaaacta
      961 ccatagatca acttcataat tcgcttaatg cagcttctaa aacgggaaat tcaaatgaag
     1021 ttttgcaaga tcctcatatt ggagatatgt acggatctgt aacgcctttg agacctcaag
     1081 ttacgagaat gttaggtaaa tatgctaaag aaaaagaaga catgttatcc ctgcgtcaag
     1141 tcttagcaaa tgcagaacgc tcatataatc aactaatgga tcgtgcagct aacgcacata
     1201 tctcaccacc agttccaggc cctgcactct acgcaggaat gactcatgct aataatactc
     1261 cagtaatgcc acctcagagg cagagttacc aaagcaatga gtactcacct tacccttcta
     1321 atttgccaat ccaacatcct acgaatagcg caaataatac cccacaatac ggttatgatc
     1381 ttggatattc agttgttagc caaccaccac ctggttatga acaaacggat ccaggaagtt
     1441 cggggtccag cggttcttca ggcgctagca tggatagcac tgagagcctt ttcactggag
     1501 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     1561 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     1621 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct
     1681 tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     1741 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     1801 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     1861 aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat
     1921 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     1981 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     2041 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     2101 ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac
     2161 atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt ttcactggag
     2221 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     2281 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     2341 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct
     2401 tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     2461 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     2521 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     2581 aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat
     2641 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     2701 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     2761 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     2821 ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac
     2881 atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt ttcactggag
     2941 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     3001 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     3061 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct
     3121 tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     3181 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     3241 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     3301 aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat
     3361 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     3421 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     3481 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     3541 ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac
     3601 atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt ttcactggag
     3661 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     3721 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     3781 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct
     3841 tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     3901 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     3961 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     4021 aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat
     4081 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     4141 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     4201 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     4261 ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac
     4321 atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt ttcactggag
     4381 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     4441 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     4501 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct
     4561 tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     4621 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     4681 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     4741 aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat
     4801 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     4861 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     4921 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     4981 ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac
     5041 atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt ttcactggag
     5101 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     5161 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     5221 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct
     5281 tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     5341 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     5401 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     5461 aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat
     5521 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     5581 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     5641 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     5701 ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac
     5761 atggcagctc cggctccagc gggtaaagcg gccgcccctt caacttatat agcaccttta
     5821 catatcttta atttttacga agagcatttt attcaacttt tgtaccttta aagacggttt
     5881 tcagcccaca cattttttgt tattttacac cacacagtaa aactttcttg cttttttttc
     5941 ttgagactcc ccgaccattg ttcgatattc caattttcgc aatcttcaga aaagggtatc
     6001 ctaacattct aaattaataa taaagaagag aggctacaag catttattag tgcggagcat
     6061 ttaaagtaac ttcaggaaat aaacgcgcaa ataaaccaaa aatgtccact gattctattg
     6121 ttaaagcctc caactggaga ttagtcgaag tcggccgtgt cgttttgatc aaaaagggtc
     6181 aatccgcagg taaattggct gctatcgttg aaattatcga ccaaaagaag gtatgttgaa
     6241 cctaaaaccc accgtggaca aactgaggag gaaattgtaa ggaagagaaa gtccccgtat
     6301 gttcaggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6361 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6421 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6481 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     6541 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6601 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6661 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6721 cgaaagggcg ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     6781 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     6841 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     6901 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     6961 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     7021 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     7081 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     7141 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     7201 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     7261 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     7321 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     7381 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     7441 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     7501 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     7561 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     7621 tcgtttcctg catgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     7681 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     7741 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     7801 agaataaaaa aaaaatgatg aattgaaaag ctggcctcgt gatacgccta tttttatagg
     7861 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     7921 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7981 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8041 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8101 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8161 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8221 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8281 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8341 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8401 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8461 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8521 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8581 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8641 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8701 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     8761 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8821 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8881 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8941 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9001 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     9721 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     9781 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
