LOCUS       sequence-438030-        8679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP-tagged marker for the vacuolar membrane (FAP optimized for yeast
            expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8679)
  AUTHORS   Oppenheimer KG, Hager NA, McAtee CK, Filiztekin E, Shang C, Warnick 
            JA, Bruchez MP, Brodsky JL, Prosser DC, Kwiatkowski AV, O'Donnell AF
  TITLE     Optimization of the fluorogen-activating protein tag for 
            quantitative protein trafficking and colocalization studies in S. 
            cerevisiae.
  JOURNAL   Mol Biol Cell. 2024 Jul 1;35(7):mr5. doi: 10.1091/mbc.E24-04-0174. 
            Epub 2024 May 29.
  PUBMED    38809589
REFERENCE   2  (bases 1 to 8679)
  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"; journalName: "Mol Biol 
            Cell."; date: "2024-07-1"; volume: "35(7):mr5. doi"; pages: " 
            10.1091/mbc.E24-04-0174. Epub 2024 May 29"
COMMENT     Sequence Label: pRS413-TEF1pr-VPH1-FAPoptim
FEATURES             Location/Qualifiers
     source          1..8679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(201..220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     320..342
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(380..398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    446..949
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        979..1083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1084..1152,1153..1944)
                     /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(1302..1321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2115..2703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2604..2623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2857..2874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2991..3012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3027..3032,3033..3050,3051..3057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3065..3081
                     /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     3070..3092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3089..3105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3089..3105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3124..3144
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3126..3144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3162..3560
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3564..3580
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3564..3580
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             6888..6917
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6918..6947
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6982..7001)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6983..7001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7011..7028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7011..7027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7020..7042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7168..7623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7255..7274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7465..7486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7883..8545)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(join(8546..8679,1..54))
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaaaaaaaaa atgaaaatca ttaccgaggc ataaaaaaat atagagtgta ctagaggagg
       61 ccaagagtaa tagaaaaaga aaattgcggg aaaggactgt gttatgactt ccctgactaa
      121 tgccgtgttc aaacgatacc tggcagtgac tcctagcgct caccaagctc ttaaaacggg
      181 aatttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      241 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      301 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      361 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      421 ataatggttt cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt
      481 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
      541 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
      601 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
      661 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
      721 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
      781 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
      841 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
      901 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc
      961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2881 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3001 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3061 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3121 cgcgcaatta accctcacta aagggaacaa aagctggagc tcatagcttc aaaatgtttc
     3181 tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa
     3241 acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac
     3301 ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa
     3361 aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt
     3421 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     3481 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     3541 gcatagcaat ctaatctaag ttttctagaa ctagtggatc catggcagag aaggaggaag
     3601 cgatttttcg ctctgctgaa atggctttag tccaattcta tattcctcaa gaaatttcaa
     3661 gagactctgc ttacacttta ggtcaattgg gtcttgttca attccgtgac ttgaactcta
     3721 aggtgcgtgc gtttcaaaga actttcgtga acgaaattag aagactggat aatgtagaaa
     3781 gacagtatcg ttacttttat tctcttttga agaaacacga tattaagctc tacgaaggag
     3841 acacggacaa atatttggac ggctcaggtg aattgtacgt tccaccaagc ggttcagtga
     3901 tagatgatta tgtccggaac gcttcatatt tggaagaaag attgattcaa atggaggatg
     3961 caaccgatca aattgaagtc cagaaaaatg acttggaaca gtatcgcttt attttgcagt
     4021 caggtgatga atttttcttg aagggtgata ataccgacag cacttcctat atggatgaag
     4081 acatgatcga cgctaatggg gaaaacattg ctgctgctat cggtgcttct gtaaactatg
     4141 tcactggtgt cattgctaga gacaaagttg ccaccttaga acaaattctt tggagagtat
     4201 taagaggtaa ccttttcttc aaaactgttg aaattgaaca acctgtttat gatgtcaaaa
     4261 ccagggagta taaacataaa aatgctttta tcgtattttc tcacggtgat ctgattatta
     4321 aaagaatcag aaagattgcg gaatcattgg atgccaatct ttacgatgtt gactcttcca
     4381 acgagggtag atcacaacaa ttggccaagg tcaacaagaa tttgagtgat ttgtacacag
     4441 ttttgaaaac cacttctacc actttagaaa gtgaattata tgccattgcc aaagaattgg
     4501 actcttggtt ccaagatgtt acccgtgaaa aggcgatttt tgaaattttg aacaagtcta
     4561 actatgatac caatagaaag attttgattg ctgaaggttg gataccaaga gacgaattgg
     4621 ctactttgca agctcgtctt ggtgaaatga tcgcaagatt gggtattgat gtcccatcca
     4681 ttatccaagt cctggataca aaccacactc cacctacctt ccacagaact aacaagttta
     4741 ctgctggttt ccaaagtatc tgtgactgtt acggtattgc tcagtacaga gaaatcaatg
     4801 ctggtttacc cacaattgtc actttccctt tcatgtttgc catcatgttt ggtgatatgg
     4861 gtcacgggtt cttaatgacc ttagccgcat tgtctcttgt attgaatgaa aagaaaatca
     4921 acaaaatgaa aagaggcgaa attttcgata tggccttcac tggtagatac attattttgt
     4981 tgatgggtgt cttttccatg tacacaggtt tcctttacaa cgatatcttc tctaaaacta
     5041 tgactatttt caagtctggt tggaaatggc ctgatcattg gaaaaaaggt gagagtatta
     5101 ctgctacatc ggtgggtaca taccctatcg gtttagattg ggcttggcat ggaactgaaa
     5161 atgctttgtt attttctaat tcttacaaaa tgaaactatc aattttaatg gggttcatcc
     5221 acatgaccta ttcttatttc ttttcgttgg ctaaccacct atactttaac tctatgattg
     5281 atatcatcgg taactttatt cctggtttgc tatttatgca aggtatcttt ggttatcttt
     5341 ccgtttgtat tgtttacaaa tgggctgttg attgggttaa ggacggaaag cctgctccag
     5401 gtttgttaaa tatgttgatc aacatgtttt tatcaccagg aactattgac gatgaattat
     5461 accctcatca agcaaaggtc caagtgtttt tgttgttgat ggccttggtt tgtattcctt
     5521 ggttgctatt ggtgaagcca ttacatttca aattcactca taaaaagaaa tctcacgaac
     5581 cactgccatc gactgaagca gatgctagtt ctgaagattt ggaagcacaa caattaattt
     5641 ccgcgatgga cgccgatgac gctgaagaag aagaagttgg ttctggatct catggtgaag
     5701 actttggtga tattatgatt catcaagtta ttcatacaat tgaattctgt ttgaattgtg
     5761 tttcgcacac tgcatcctat ttacgtttat gggccttatc attggcacat gctcaattgt
     5821 ctagtgtttt atggacaatg acaattcaaa ttgcctttgg atttagagga tttgtgggtg
     5881 tgtttatgac ggttgcactt tttgccatgt ggttcgcact aacatgtgca gttcttgttt
     5941 tgatggaagg tacatctgcc atgcttcatt ccttacgttt gcactgggtt gaatctatgt
     6001 ccaagttttt cgtgggtgaa ggtttaccat acgaaccatt cgcatttgag tataaagaca
     6061 tggaagtcgc tgttgctagt gcaagctctt ccgcttcaag ccccgggctg caggaattcg
     6121 atatcaagct tatcgatacc gtcgacgcgg gtgcgggtgc gggtatgcaa gctgttgtta
     6181 ctcaagaacc atctgttact gtttctccag gtggtactgt tatcttgact tgtggttctg
     6241 gtactggtgc tgttacttct ggtcactacg ctaactggtt ccaacaaaag ccaggtcaag
     6301 ctccaagagc tttgatcttc gacactgaca agaagtactc ttggactcca ggtagattct
     6361 ctggttcttt gttgggtgct aaggctgctt tgactatctc tgacgctcaa ccagaagacg
     6421 aagctgaata ctactgttct ttgtctgacg ttgacggtta cttgttcggt ggtggtactc
     6481 aattgactgt tttgtctggt ggtggtggtt ctggtggtgg tggttctggt ggtggtggtt
     6541 ctggtggtgg tggttctcaa gctgttgtta ctcaagaacc atctgttact gtttctccag
     6601 gtggtactgt tatcttgact tgtggttctg gtactggtgc tgttacttct ggtcactacg
     6661 ctaactggtt ccaacaaaag ccaggtcaag ctccaagagc tttgatcttc gacactgaca
     6721 agaagtactc ttggactcca ggtagattct ctggttcttt gttgggtgct aaggctgctt
     6781 tgactatctc tgacgctcaa ccagaagacg aagctgaata ctactgttct ttgtctgacg
     6841 ttgacggtta cttgttcggt ggtggtactc aattgactgt tttgtctgag cagaagctga
     6901 tcagcgagga ggacctggag cagaagctga tcagcgagga ggacctgtaa cgcctcgagg
     6961 gggggcccgg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     7021 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     7081 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     7141 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
     7201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     7261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7381 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7501 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7561 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     7621 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagatcc
     7681 gtcgagttca agagaaaaaa aaagaaaaag caaaaagaaa aaaggaaagc gcgcctcgtt
     7741 cagaatgaca cgtatagaat gatgcattac cttgtcatct tcagtatcat actgttcgta
     7801 tacatactta ctgacattca taggtataca tatatacaca tgtatatata tcgtatgctg
     7861 cagctttaaa taatcggtgt cactacataa gaacaccttt ggtggaggga acatcgttgg
     7921 taccattggg cgaggtggct tctcttatgg caaccgcaag agccttgaac gcactctcac
     7981 tacggtgatg atcattcttg cctcgcagac aatcaacgtg gagggtaatt ctgctagcct
     8041 ctgcaaagct ttcaagaaaa tgcgggatca tctcgcaaga gagatctcct actttctccc
     8101 tttgcaaacc aagttcgaca actgcgtacg gcctgttcga aagatctacc accgctctgg
     8161 aaagtgcctc atccaaaggc gcaaatcctg atccaaacct ttttactcca cgcacggccc
     8221 ctagggcctc tttaaaagct tgaccgagag caatcccgca gtcttcagtg gtgtgatggt
     8281 cgtctatgtg taagtcacca atgcactcaa cgattagcga ccagccggaa tgcttggcca
     8341 gagcatgtat catatggtcc agaaacccta tacctgtgtg gacgttaatc acttgcgatt
     8401 gtgtggcctg ttctgctact gcttctgcct ctttttctgg gaagatcgag tgctctatcg
     8461 ctaggggacc accctttaaa gagatcgcaa tctgaatctt ggtttcattt gtaatacgct
     8521 ttactagggc tttctgctct gtcatctttg ccttcgttta tcttgcctgc tcatttttta
     8581 gtatattctt cgaagaaatc acattacttt atataatgta taattcatta tgtgataatg
     8641 ccaatcgcta agaaaaaaaa agagtcatcc gctaggtgg
//
