LOCUS       sequence-439393-        8507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP insert for N-terminally tagging genes of interest (optimized FAP
            for yeast expression + 2xMYC tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8507)
  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 8507)
  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: pRS415-ADH1pr-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..8507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(168..187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     287..309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(347..365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    407..910
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        947..1051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1052..1120,1121..1912)
                     /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(1270..1289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2083..2671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2572..2591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2825..2842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2959..2980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2995..3000,3001..3018,3019..3025)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3033..3049
                     /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     3038..3060
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3057..3073
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3057..3073
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3092..3112
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3094..3112
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3881..4585
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     4599..4615
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4599..4615
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5340..5369
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5370..5399
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5453..5469)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5454..5470)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5472..5719
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5488..5506)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5737..5756)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5738..5756)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5766..5783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5766..5782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5775..5797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5923..6378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6010..6029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6220..6241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6679..7086
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7087..8181
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac ctgaccattt gatggagttt
       61 aagtcaatac cttcttgaac catttcccat aatggtgaaa gttccctcaa gaattttact
      121 ctgtcagaaa cggccttaac gacgtagtcg atatggtgca ctctcagtac aatctgctct
      181 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      241 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
      301 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
      361 ctatttttat aggttaatgt catgataata atggtttctt aggacggatc gcttgcctgt
      421 aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt
      481 atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta
      541 cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta
      601 catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa
      661 gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa
      721 ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc
      781 cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt
      841 tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat
      901 agcacgtgat gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
      961 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1021 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1081 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1141 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1201 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1261 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1321 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1381 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1441 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1501 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1561 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1621 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1681 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1741 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     1801 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1861 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1921 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1981 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2041 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2101 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2161 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2221 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2281 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2341 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2401 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2461 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2521 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2581 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2641 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2701 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2761 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2821 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     2881 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2941 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     3001 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     3061 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     3121 gctggagctc gccgggatcg aagaaatgat ggtaaatgaa ataggaaatc aaggagcatg
     3181 aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt tgatatgatg
     3241 tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca tgctgactct
     3301 gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc tgtgcccggc
     3361 ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg agattggaga
     3421 agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg gtgtcattat
     3481 ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa gaatgagcca
     3541 agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat gaccttgaag
     3601 gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg gttgctacca
     3661 gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt acgctttcaa
     3721 ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac acttctccta
     3781 tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac ccctccgcgc
     3841 tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt tgtttccggg
     3901 tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga ttcctataat
     3961 accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat agaacagata
     4021 ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc attgatggtg
     4081 gtacataacg aactaatact gtagccctag acttgatagc catcatcata tcgaagtttc
     4141 actacccttt ttccatttgc catctattga agtaataata ggcgcatgca acttcttttc
     4201 tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa
     4261 tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt
     4321 tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc
     4381 acactactct ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat
     4441 aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat taatcttttg
     4501 tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca caatatttca
     4561 agctatacca agcatacaat caactccaag ctggccgctc tagaactagt ggatccatgc
     4621 aagctgttgt tactcaagaa ccatctgtta ctgtttctcc aggtggtact gttatcttga
     4681 cttgtggttc tggtactggt gctgttactt ctggtcacta cgctaactgg ttccaacaaa
     4741 agccaggtca agctccaaga gctttgatct tcgacactga caagaagtac tcttggactc
     4801 caggtagatt ctctggttct ttgttgggtg ctaaggctgc tttgactatc tctgacgctc
     4861 aaccagaaga cgaagctgaa tactactgtt ctttgtctga cgttgacggt tacttgttcg
     4921 gtggtggtac tcaattgact gttttgtctg gtggtggtgg ttctggtggt ggtggttctg
     4981 gtggtggtgg ttctggtggt ggtggttctc aagctgttgt tactcaagaa ccatctgtta
     5041 ctgtttctcc aggtggtact gttatcttga cttgtggttc tggtactggt gctgttactt
     5101 ctggtcacta cgctaactgg ttccaacaaa agccaggtca agctccaaga gctttgatct
     5161 tcgacactga caagaagtac tcttggactc caggtagatt ctctggttct ttgttgggtg
     5221 ctaaggctgc tttgactatc tctgacgctc aaccagaaga cgaagctgaa tactactgtt
     5281 ctttgtctga cgttgacggt tacttgttcg gtggtggtac tcaattgact gttttgtctg
     5341 agcagaagct gatcagcgag gaggacctgg agcagaagct gatcagcgag gaggacctgg
     5401 cgggtgcggg tgcgggtcgc cccgggctgc aggaattcga tatcaagctt atcgataccg
     5461 tcgacctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     5521 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     5581 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     5641 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     5701 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     5761 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5821 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5881 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     5941 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6001 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6061 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6121 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6181 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6241 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6301 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6361 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6421 acaccgcata tcgaccggtc gaggagaact tctagtatat ctacatacct aatattattg
     6481 ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat
     6541 tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac
     6601 tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga
     6661 aatatcttga ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga
     6721 atctcttagc aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg
     6781 cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat
     6841 ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct
     6901 tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg
     6961 acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt
     7021 ctaacttttc ttacctttta catttcagca atatatatat atatatttca aggatatacc
     7081 attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa
     7141 atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag
     7201 ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt
     7261 ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt
     7321 ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa
     7381 gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac
     7441 ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta
     7501 gtgggaggta tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat
     7561 agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc
     7621 ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct
     7681 tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag
     7741 gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta
     7801 aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt
     7861 atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac
     7921 accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag
     7981 gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg
     8041 cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga
     8101 actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa
     8161 gttaagaaaa tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt
     8221 ataaatgaaa ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt
     8281 agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta
     8341 aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc
     8401 actttaacat taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa
     8461 aatcatgaaa ctatgattcc taatttatat attggaggat tttctct
//
