LOCUS       sequence-199757-        9714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Apl2 with Apl2-iGFPx6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9714)
  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 9714)
  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-APL2-iGFPx6
FEATURES             Location/Qualifiers
     source          1..9714
                     /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(6207..6224)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6207..6223)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6216..6238)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6432..6451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6551..6573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6691..7494)
                     /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(7495..7715)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7722..7740)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7808..7912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7913..7981,7982..8773)
                     /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(8131..8150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8944..9532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9433..9452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9686..9703
                     /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 gtcgatcctt ccaacttgaa acaatgtact ttgttgttga
      301 cagaattaaa ggaatacgca atggaatatg aacctgagtt tgtttcaaga gctattcaag
      361 ctttgtccca gttgggtatt aaatatgcac aagaatcatt cgtcagtaaa gtcttggata
      421 ttctcctaga attgttagaa agacaagaca caataaaaga cgactgctgt atatcattat
      481 gtgatttatt aagacattgc cccggaaacg ataagatggc caaacaagtt tgtgcagtat
      541 ttaatacttg gtcaaatcca gaggtgttat tacaatcaga tattgcgaag tgcaactatg
      601 tctggttgct aggacagcat cccaacaatt tctcagattt ggagtcaaaa atcaatattt
      661 ttatagagaa ctttgttcaa gaggaagctt taacacagat gtctttacta atgacaatag
      721 tcagactaca tgctacttta acgggtagta tgctacaaag cgtcctagaa ttggccacac
      781 aacagacaca tgaattagat gtgcgagata tggctatgat gtattggaga tgcctttcga
      841 tgccaaataa tgaaagtttg gttaatgatc tatgtcaaaa taaacttcca atgatctcca
      901 atacactgga gaagttttca ccagaagtgc tagagaaact attaatggaa ttgggtacta
      961 taagttccat ttatttcaag cccgactcaa ataggagaaa aggcaaaaag tatgttcaaa
     1021 atatagttaa aggaaaacat atagaggaat tagaaagcat ggcaaaaaat gaaatttcta
     1081 gcaaggctaa cgatgatgtg ctattggatt ttgatgaaag agatgatgta acaaatacaa
     1141 atgcaggaat gctgaatact ttaacaactt taggcgattt ggatgattta tttgatttcg
     1201 gaccatctga ggatgccaca caaataaata caaatgatac caaggctgta caagggttga
     1261 aggagctgaa actaggtggc gactcaaacg gtatatcttc tggtggtaaa aacaatcccg
     1321 atgtttcggg cggcaatata gtgtcacagg atctcctcga tttattcacg gatccaggaa
     1381 gttcggggtc cagcggttct tcaggcgcta gcatggatag cactgagagc cttttcactg
     1441 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     1501 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     1561 ctggaaaact acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcagt
     1621 gcttttcaag atacccggat catatgaaac agcatgactt tttcaagagt gccatgcccg
     1681 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     1741 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     1801 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     1861 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     1921 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     1981 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     2041 atcccaacga aaagagagac cacatggtcc ttaaagagtt tgtaacagct gctgggatta
     2101 cacatggcag ctccggctcc agcgggtcta gcatggatag cactgagagc cttttcactg
     2161 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     2221 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     2281 ctggaaaact acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcagt
     2341 gcttttcaag atacccggat catatgaaac agcatgactt tttcaagagt gccatgcccg
     2401 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     2461 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     2521 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     2581 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     2641 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     2701 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     2761 atcccaacga aaagagagac cacatggtcc ttaaagagtt tgtaacagct gctgggatta
     2821 cacatggcag ctccggctcc agcgggtcta gcatggatag cactgagagc cttttcactg
     2881 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     2941 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     3001 ctggaaaact acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcagt
     3061 gcttttcaag atacccggat catatgaaac agcatgactt tttcaagagt gccatgcccg
     3121 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     3181 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     3241 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     3301 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     3361 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     3421 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     3481 atcccaacga aaagagagac cacatggtcc ttaaagagtt tgtaacagct gctgggatta
     3541 cacatggcag ctccggctcc agcgggtcta gcatggatag cactgagagc cttttcactg
     3601 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     3661 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     3721 ctggaaaact acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcagt
     3781 gcttttcaag atacccggat catatgaaac agcatgactt tttcaagagt gccatgcccg
     3841 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     3901 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     3961 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     4021 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     4081 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     4141 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4201 atcccaacga aaagagagac cacatggtcc ttaaagagtt tgtaacagct gctgggatta
     4261 cacatggcag ctccggctcc agcgggtcta gcatggatag cactgagagc cttttcactg
     4321 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     4381 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     4441 ctggaaaact acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcagt
     4501 gcttttcaag atacccggat catatgaaac agcatgactt tttcaagagt gccatgcccg
     4561 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     4621 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     4681 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     4741 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     4801 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     4861 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4921 atcccaacga aaagagagac cacatggtcc ttaaagagtt tgtaacagct gctgggatta
     4981 cacatggcag ctccggctcc agcgggtcta gcatggatag cactgagagc cttttcactg
     5041 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     5101 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     5161 ctggaaaact acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcagt
     5221 gcttttcaag atacccggat catatgaaac agcatgactt tttcaagagt gccatgcccg
     5281 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     5341 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     5401 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     5461 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     5521 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     5581 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     5641 atcccaacga aaagagagac cacatggtcc ttaaagagtt tgtaacagct gctgggatta
     5701 cacatggcag ctccggctcc agcgggtaaa gcggccgcct ataaacgtcc gttgtagtga
     5761 acttaagtat tataccatac atatataccc ttcgtattaa tttttccttt ttattgttgg
     5821 gctgtgtgta tataatttat gtacaagaat ggatatatca aaaaatcctt aaaagatgta
     5881 aatcaaaaac atcaatcaac acagcttatt ttgtagagct ttgtttttga cataactttt
     5941 caagcttgtt gattttttct aaaacaattt tatccctttc accatctaaa tatttctcat
     6001 ctgtttgata aaacaggagg cacgatttca tttcatctcg ttgcttatca caagtgggaa
     6061 aaactttcat tccatttgcc ttagcacaat ctgccattgc ttgaacaaaa tttgcacatt
     6121 ttttgtatgt ttccgtcttc aagttcttgc gagcctgttg ttcttcgcga ggacttagca
     6181 cccatatagg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     6241 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6301 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6361 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     6421 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6481 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6541 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6601 agacgaaagg gcgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     6661 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     6721 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     6781 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
     6841 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
     6901 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     6961 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     7021 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     7081 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     7141 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     7201 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     7261 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     7321 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     7381 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     7441 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     7501 tcttcgtttc ctgcatgttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     7561 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     7621 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa
     7681 aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct ggcctcgtga tacgcctatt
     7741 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     7801 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7861 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7921 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7981 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8041 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8101 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8161 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8221 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8281 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8341 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8401 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     8461 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     8521 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8581 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     8641 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8701 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8761 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8821 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8881 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     9121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     9421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     9481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     9541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     9601 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     9661 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
