LOCUS       sequence-438159-        8913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP-tagged marker for the plasma membrane (FAP optimized for yeast 
            expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8913)
  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 8913)
  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-PMA1-FAPoptim
FEATURES             Location/Qualifiers
     source          1..8913
                     /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             7122..7151
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7152..7181
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(7216..7235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7217..7235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7245..7262)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7245..7261)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7254..7276)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7402..7857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7489..7508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7699..7720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8117..8779)
                     /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(8780..8913,1..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 catgactgat acatcatcct
     3601 cttcatcatc ctcttcagca tcttctgttt cagctcatca gccaactcaa gaaaagcctg
     3661 ctaagactta cgatgacgct gcatctgaat cttctgacga tgacgatatc gatgctttaa
     3721 tcgaagaact acaatctaat cacggtgtcg acgacgaaga cagtgataac gatggtccag
     3781 ttgccgccgg tgaagctaga ccagttccag aagaatattt acaaactgac ccatcttacg
     3841 gtttaacttc cgatgaagtt ttgaaaagaa gaaagaagta cggtttgaat caaatggctg
     3901 acgaaaaaga atctttagtc gttaagttcg ttatgttttt cgtcggtcca attcaattcg
     3961 ttatggaagc cgctgctatt ttggctgccg gtttgtccga ttgggtcgat ttcggtgtta
     4021 tctgtggttt gttaatgtta aacgctggtg ttggtttcgt tcaagaattc caagctggtt
     4081 ctattgtcga tgaattgaaa aagactttgg ctaacactgc tgttgttatc agagacggtc
     4141 aattggttga aattccagcc aacgaagtcg tcccaggtga tattttgcaa ttggaagatg
     4201 gtactgttat cccaactgat ggtcgtattg tcactgaaga ctgtttcttg caaatcgatc
     4261 aatctgctat tactggtgaa tctttggctg tcgacaaaca ttacggtgac caaactttct
     4321 cttcttccac tgttaagaga ggtgaaggtt tcatggttgt taccgctacc ggtgacaaca
     4381 ctttcgttgg tagagctgct gctttggtta acaaagccgc tggtggtcaa ggtcatttca
     4441 ctgaagtttt gaacggtatt ggtattatct tattggtttt ggtcattgcc actttgttgt
     4501 tggtctggac tgcttgtttc tacagaacca acggtattgt tagaatcttg agatacactc
     4561 taggtattac tattattggt gtcccagtcg gtttgccagc tgtcgttacc accactatgg
     4621 ccgtcggtgc tgcttacttg gctaagaaac aagccattgt tcaaaagttg tctgctattg
     4681 aatccttggc tggtgtcgaa atcttgtgtt ccgacaaaac cggtactttg accaagaaca
     4741 agttgtcctt gcacgaacca tacactgttg aaggtgtttc tccagacgac ttgatgttga
     4801 ctgcttgttt ggctgcttcc agaaagaaga agggtttgga tgctattgat aaggctttct
     4861 tgaagtcttt gaagcaatat ccaaaggcta aggacgcttt gaccaagtac aaggttttgg
     4921 aattccatcc atttgaccct gtctccaaga aggttactgc cgttgtcgaa tctccagaag
     4981 gtgaaagaat tgtttgtgtt aagggtgctc cattattcgt cttgaagact gtcgaagaag
     5041 atcacccaat cccagaagat gtccatgaaa actacgaaaa caaggttgct gaattggctt
     5101 ctagagggtt ccgtgcttta ggtgttgcta gaaagagagg tgaaggtcac tgggaaatct
     5161 tgggtgttat gccatgtatg gatccaccaa gagacgatac tgttcaaact gttagcgaag
     5221 ctagacactt gggtctaaga gttaagatgt taactggtga tgctgttggt attgctaagg
     5281 aaacttgtag acaattgggt ttgggtacca acatttacaa cgctgaaaga ttaggtctag
     5341 gtggcggtgg tgacatgcca ggttctgaat tagctgattt tgttgaaaat gccgatggtt
     5401 tcgctgaagt tttcccacaa cataaataca gagttgttga aatcttgcaa aacagaggtt
     5461 acttggttgc tatgactggt gatggtgtta acgatgctcc atctttgaag aaggctgata
     5521 ctggtattgc tgtcgaaggt gctactgatg ctgccagatc tgctgctgat attgttttct
     5581 tggctcctgg tctatctgct attattgatg ctttgaagac ctccagacaa atttttcaca
     5641 gaatgtactc ttacgttgtt taccgtattg ctttgtctct acatttggaa atcttcttgg
     5701 gtctatggat tgctattttg gataactctt tggacattga tttgattgtt ttcatcgcta
     5761 ttttcgctga tgttgctact ttggctattg cttacgataa tgctccttac tctccaaagc
     5821 ccgttaaatg gaacctacca agattatggg gtatgtctat tattttgggc atagttttag
     5881 ctataggttc ttggattacc ttgactacta tgttcttacc aaagggtggt attatccaaa
     5941 acttcggtgc tatgaacggt attatgttct tgcaaatttc cttgactgaa aactggttga
     6001 ttttcattac cagagctgct ggtccattct ggtcttctat cccatcctgg caattggctg
     6061 gtgccgtctt cgctgtcgac atcatcgcta ccatgtttac cttattcggt tggtggtctg
     6121 aaaactggac tgatattgtt actgtcgtcc gtgtctggat ctggtctatc ggtatcttct
     6181 gtgttttggg tggtttctac tacgaaatgt ccacttctga agcctttgac agattgatga
     6241 acggtaagcc aatgaaggaa aagaagtcta ccagaagtgt cgaagacttc atggctgcta
     6301 tgcaaagagt ctctactcaa cacgaaaagg aaacccccgg gctgcaggaa ttcgatatca
     6361 agcttatcga taccgtcgac gcgggtgcgg gtgcgggtat gcaagctgtt gttactcaag
     6421 aaccatctgt tactgtttct ccaggtggta ctgttatctt gacttgtggt tctggtactg
     6481 gtgctgttac ttctggtcac tacgctaact ggttccaaca aaagccaggt caagctccaa
     6541 gagctttgat cttcgacact gacaagaagt actcttggac tccaggtaga ttctctggtt
     6601 ctttgttggg tgctaaggct gctttgacta tctctgacgc tcaaccagaa gacgaagctg
     6661 aatactactg ttctttgtct gacgttgacg gttacttgtt cggtggtggt actcaattga
     6721 ctgttttgtc tggtggtggt ggttctggtg gtggtggttc tggtggtggt ggttctggtg
     6781 gtggtggttc tcaagctgtt gttactcaag aaccatctgt tactgtttct ccaggtggta
     6841 ctgttatctt gacttgtggt tctggtactg gtgctgttac ttctggtcac tacgctaact
     6901 ggttccaaca aaagccaggt caagctccaa gagctttgat cttcgacact gacaagaagt
     6961 actcttggac tccaggtaga ttctctggtt ctttgttggg tgctaaggct gctttgacta
     7021 tctctgacgc tcaaccagaa gacgaagctg aatactactg ttctttgtct gacgttgacg
     7081 gttacttgtt cggtggtggt actcaattga ctgttttgtc tgagcagaag ctgatcagcg
     7141 aggaggacct ggagcagaag ctgatcagcg aggaggacct gtaacgcctc gagggggggc
     7201 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     7261 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     7321 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     7381 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     7441 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     7501 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     7561 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     7621 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     7681 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     7741 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     7801 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     7861 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag atccgtcgag
     7921 ttcaagagaa aaaaaaagaa aaagcaaaaa gaaaaaagga aagcgcgcct cgttcagaat
     7981 gacacgtata gaatgatgca ttaccttgtc atcttcagta tcatactgtt cgtatacata
     8041 cttactgaca ttcataggta tacatatata cacatgtata tatatcgtat gctgcagctt
     8101 taaataatcg gtgtcactac ataagaacac ctttggtgga gggaacatcg ttggtaccat
     8161 tgggcgaggt ggcttctctt atggcaaccg caagagcctt gaacgcactc tcactacggt
     8221 gatgatcatt cttgcctcgc agacaatcaa cgtggagggt aattctgcta gcctctgcaa
     8281 agctttcaag aaaatgcggg atcatctcgc aagagagatc tcctactttc tccctttgca
     8341 aaccaagttc gacaactgcg tacggcctgt tcgaaagatc taccaccgct ctggaaagtg
     8401 cctcatccaa aggcgcaaat cctgatccaa acctttttac tccacgcacg gcccctaggg
     8461 cctctttaaa agcttgaccg agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta
     8521 tgtgtaagtc accaatgcac tcaacgatta gcgaccagcc ggaatgcttg gccagagcat
     8581 gtatcatatg gtccagaaac cctatacctg tgtggacgtt aatcacttgc gattgtgtgg
     8641 cctgttctgc tactgcttct gcctcttttt ctgggaagat cgagtgctct atcgctaggg
     8701 gaccaccctt taaagagatc gcaatctgaa tcttggtttc atttgtaata cgctttacta
     8761 gggctttctg ctctgtcatc tttgccttcg tttatcttgc ctgctcattt tttagtatat
     8821 tcttcgaaga aatcacatta ctttatataa tgtataattc attatgtgat aatgccaatc
     8881 gctaagaaaa aaaaagagtc atccgctagg tgg
//
