LOCUS       sequence-202432-        9070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Apl5 with Apl5-iGFPx6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9070)
  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 9070)
  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-APL5-iGFPx6
FEATURES             Location/Qualifiers
     source          1..9070
                     /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(5568..5585)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5568..5584)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5577..5599)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5793..5812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5912..5934
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6052..6855)
                     /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(6856..7071)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7078..7096)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7164..7268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7269..7337,7338..8129)
                     /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(7487..7506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8300..8888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8789..8808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9042..9059
                     /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 atgccatttg agattgagag agaaaaggaa cggatgtcaa atccttatta tctaggtgaa
      301 gaagacgaag aaagaactaa gaactctaaa gatttgttgg acttgaatga ggaggaaagt
      361 agtgataaaa agccagaaac tattagatta aatagaaccg ataattcgtt gaattctcta
      421 agtttatcta caactgaaat cagcagaaag aagaagaagg gaaagaagaa aaatagggtt
      481 caagtcttat ccgatgaacc tgttatcgaa gccgctccca aaaggaaaga tgcattccag
      541 aaaccccatg ataatcattc aactcaaaat cctttgaaaa aggacaagat taatctgaga
      601 atgcactctc aacttgaaaa ttttgatttt tccaattttg ggcaatcgag taacgcagga
      661 agaggatcgc aggaagaggg caaccttaga aaggaagacg agttagaatt aagtcgttta
      721 gaggccaatc ttattgtaaa agatgaaaag gataatttaa gtgatactga agaagttata
      781 gttataaaaa agaagaagaa agggaaaaag tcaaagtcaa aaaacaaact aaaaacgaaa
      841 gcaaaaaatt ctccagaacc aaatgaattt cttcgagacc aaagcactga cattacggat
      901 ccaggaagtt cggggtccag cggttcttca ggcgctagca tggatagcac tgagagcctt
      961 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1021 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1081 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     1141 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     1201 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1261 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1321 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     1381 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     1441 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     1501 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     1561 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     1621 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     1681 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1741 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1801 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     1861 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     1921 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1981 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     2041 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     2101 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     2161 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     2221 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     2281 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     2341 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     2401 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     2461 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     2521 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     2581 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     2641 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     2701 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     2761 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     2821 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     2881 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     2941 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     3001 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     3061 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     3121 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     3181 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     3241 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     3301 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     3361 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     3421 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     3481 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     3541 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     3601 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     3661 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     3721 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     3781 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     3841 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     3901 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     3961 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     4021 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     4081 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     4141 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     4201 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     4261 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     4321 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     4381 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     4441 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     4501 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     4561 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     4621 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     4681 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     4741 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     4801 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     4861 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     4921 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     4981 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     5041 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     5101 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     5161 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     5221 gggattacac atggcagctc cggctccagc gggtaaagcg gccgccgata aaaaaatcga
     5281 agtcgtgaat tctcaatgac agcgattacg tacgtaaact ataattaatt taagtatttt
     5341 attctgcata gtatattatt gcggtgcact caatttgccg aaagagacgg catatttaat
     5401 aatgacgcga atgaatcaaa ggccgccaaa ggtaatcata gaccagagat gatttgcaaa
     5461 attatttgac atactattgc tagtgtaaca ataacacagc ataactttgc ttagacatat
     5521 atgtcattta aggcaactat caccgagtcg gtaccgagct cgaattcact ggccgtcgtt
     5581 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5641 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5701 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5761 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5821 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5881 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5941 ccgtcatcac cgaaacgcgc gagacgaaag ggcgggtaat aactgatata attaaattga
     6001 agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg
     6061 ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac
     6121 cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt
     6181 agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa
     6241 acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct
     6301 ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt
     6361 agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc
     6421 tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc
     6481 caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga
     6541 gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa
     6601 attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt
     6661 caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc
     6721 cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg
     6781 catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata
     6841 tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt
     6901 taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct
     6961 aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt
     7021 tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctggcc
     7081 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     7141 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     7201 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     7261 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     7321 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7381 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7441 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7501 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7561 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7621 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7681 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7741 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7801 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7861 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7921 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7981 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     8041 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     8101 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     8161 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     8221 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     8341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8941 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     9001 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     9061 aagcggaaga
//
