LOCUS       sequence-199812-        9038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Apl5 with 
            Apl5-mCherry2Bx6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9038)
  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 9038)
  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-mCherry2Bx6
FEATURES             Location/Qualifiers
     source          1..9038
                     /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(5531..5548)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5531..5547)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5540..5562)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5756..5775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5875..5897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6015..6818)
                     /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(6819..7039)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7046..7064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7132..7236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7237..7305,7306..8097)
                     /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(7455..7474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8268..8856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8757..8776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9010..9027
                     /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 tgagagcggc
      961 tccaccgagt ccgtcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1021 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1081 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1141 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     1201 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1261 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1321 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gactatgggc
     1381 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     1441 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     1501 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     1561 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     1621 ggctcccagg gatctagcgg cagcagttct agcatggata gcactgagag cggctccacc
     1681 gagtccgtca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1741 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1801 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     1861 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     1921 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     1981 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2041 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagactat gggctgggag
     2101 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2161 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2221 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2281 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccggctcc
     2341 cagggatcta gcggcagcag ttctagcatg gatagcactg agagcggctc caccgagtcc
     2401 gtcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2461 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2521 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2581 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2641 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2701 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2761 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ctatgggctg ggaggcctcc
     2821 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2881 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2941 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     3001 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccgg ctcccaggga
     3061 tctagcggca gcagttctag catggatagc actgagagcg gctccaccga gtccgtcatc
     3121 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     3181 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     3241 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     3301 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     3361 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     3421 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     3481 ttcccctccg acggccccgt aatgcagaag aagactatgg gctgggaggc ctcctccgag
     3541 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     3601 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     3661 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     3721 accatcgtgg aacagtacga acgcgccgag ggccgccact ccggctccca gggatctagc
     3781 ggcagcagtt ctagcatgga tagcactgag agcggctcca ccgagtccgt catcaaggag
     3841 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     3901 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3961 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     4021 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     4081 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     4141 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     4201 tccgacggcc ccgtaatgca gaagaagact atgggctggg aggcctcctc cgagcggatg
     4261 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     4321 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     4381 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     4441 gtggaacagt acgaacgcgc cgagggccgc cactccggct cccagggatc tagcggcagc
     4501 agttctagca tggatagcac tgagagcggc tccaccgagt ccgtcatcaa ggagttcatg
     4561 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     4621 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     4681 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     4741 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     4801 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     4861 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     4921 ggccccgtaa tgcagaagaa gactatgggc tgggaggcct cctccgagcg gatgtacccc
     4981 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     5041 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     5101 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     5161 cagtacgaac gcgccgaggg ccgccactcc ggctcccagg gatctagcgg cagcagttaa
     5221 gcggccgccg ataaaaaaat cgaagtcgtg aattctcaat gacagcgatt acgtacgtaa
     5281 actataatta atttaagtat tttattctgc atagtatatt attgcggtgc actcaatttg
     5341 ccgaaagaga cggcatattt aataatgacg cgaatgaatc aaaggccgcc aaaggtaatc
     5401 atagaccaga gatgatttgc aaaattattt gacatactat tgctagtgta acaataacac
     5461 agcataactt tgcttagaca tatatgtcat ttaaggcaac tatcaccgag tcggtaccga
     5521 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5581 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5641 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     5701 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     5761 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5821 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5881 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcgggt
     5941 aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact
     6001 tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct
     6061 tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca
     6121 acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc
     6181 aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct
     6241 tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc
     6301 ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac
     6361 aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc
     6421 aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct
     6481 gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat
     6541 tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact
     6601 gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg
     6661 ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca
     6721 cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga
     6781 tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcat
     6841 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
     6901 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     6961 gagattaccg aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa
     7021 aatgatgaat tgaattgaaa agctggcctc gtgatacgcc tatttttata ggttaatgtc
     7081 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7141 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7201 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7261 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7321 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7381 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7441 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7501 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7561 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7621 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7681 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7741 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7801 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7861 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7921 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7981 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8041 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8101 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8161 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8401 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8881 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8941 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     9001 accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
