LOCUS       sequence-439394-        7425 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 7425)
  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 7425)
  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-MET25pr-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..7425
                     /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        3163..3512
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3513..3529
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3513..3529
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4254..4283
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4284..4313
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4371..4387)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4372..4388)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4390..4637
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4406..4424)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4655..4674)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4656..4674)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4684..4701)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4684..4700)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4693..4715)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4841..5296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4928..4947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5138..5159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5597..6004
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6005..7099
                     /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 cggatgcaag ggttcgaatc ccttagctct cattattttt tgctttttct
     3181 cttgaggtca catgatcgca aaatggcaaa tggcacgtga agctgtcgat attggggaac
     3241 tgtggtggtt ggcaaatgac taattaagtt agtcaaggcg ccatcctcat gaaaactgtg
     3301 taacataata accgaagtgt cgaaaaggtg gcaccttgtc caattgaaca cgctcgatga
     3361 aaaaaataag atatatataa ggttaagtaa agcgtctgtt agaaaggaag tttttccttt
     3421 ttcttgctct cttgtctttt catctactat ttccttcgtg taatacaggg tcgtcagata
     3481 catagataca attctattac ccccatccat actctagaac tagtggatcc atgcaagctg
     3541 ttgttactca agaaccatct gttactgttt ctccaggtgg tactgttatc ttgacttgtg
     3601 gttctggtac tggtgctgtt acttctggtc actacgctaa ctggttccaa caaaagccag
     3661 gtcaagctcc aagagctttg atcttcgaca ctgacaagaa gtactcttgg actccaggta
     3721 gattctctgg ttctttgttg ggtgctaagg ctgctttgac tatctctgac gctcaaccag
     3781 aagacgaagc tgaatactac tgttctttgt ctgacgttga cggttacttg ttcggtggtg
     3841 gtactcaatt gactgttttg tctggtggtg gtggttctgg tggtggtggt tctggtggtg
     3901 gtggttctgg tggtggtggt tctcaagctg ttgttactca agaaccatct gttactgttt
     3961 ctccaggtgg tactgttatc ttgacttgtg gttctggtac tggtgctgtt acttctggtc
     4021 actacgctaa ctggttccaa caaaagccag gtcaagctcc aagagctttg atcttcgaca
     4081 ctgacaagaa gtactcttgg actccaggta gattctctgg ttctttgttg ggtgctaagg
     4141 ctgctttgac tatctctgac gctcaaccag aagacgaagc tgaatactac tgttctttgt
     4201 ctgacgttga cggttacttg ttcggtggtg gtactcaatt gactgttttg tctgagcaga
     4261 agctgatcag cgaggaggac ctggagcaga agctgatcag cgaggaggac ctggcgggtg
     4321 cgggtgcggg tcgccccccc cgggctgcag gaattcgata tcaagcttat cgataccgtc
     4381 gacctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     4441 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     4501 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     4561 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     4621 cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     4681 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4741 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4801 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     4861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5041 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5161 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5221 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5281 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5341 accgcatatc gaccggtcga ggagaacttc tagtatatct acatacctaa tattattgcc
     5401 ttattaaaaa tggaatccca acaattacat caaaatccac attctcttca aaatcaattg
     5461 tcctgtactt ccttgttcat gtgtgttcaa aaacgttata tttataggat aattatactc
     5521 tatttctcaa caagtaattg gttgtttggc cgagcggtct aaggcgcctg attcaagaaa
     5581 tatcttgacc gcagttaact gtgggaatac tcaggtatcg taagatgcaa gagttcgaat
     5641 ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg cgctatcgca
     5701 cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc tgacgcatat
     5761 acctttttca actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt
     5821 tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact tgaagttgac
     5881 aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt cttactttct
     5941 aacttttctt accttttaca tttcagcaat atatatatat atatttcaag gatataccat
     6001 tctaatgtct gcccctaaga agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat
     6061 cacagccgaa gccattaagg ttcttaaagc tatttctgat gttcgttcca atgtcaagtt
     6121 cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat gctacaggtg ttccacttcc
     6181 agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg
     6241 tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga
     6301 acttcaattg tacgccaact taagaccatg taactttgca tccgactctc ttttagactt
     6361 atctccaatc aagccacaat ttgctaaagg tactgacttc gttgttgtca gagaattagt
     6421 gggaggtatt tactttggta agagaaagga agacgatggt gatggtgtcg cttgggatag
     6481 tgaacaatac accgttccag aagtgcaaag aatcacaaga atggccgctt tcatggccct
     6541 acaacatgag ccaccattgc ctatttggtc cttggataaa gctaatgttt tggcctcttc
     6601 aagattatgg agaaaaactg tggaggaaac catcaagaac gaattcccta cattgaaggt
     6661 tcaacatcaa ttgattgatt ctgccgccat gatcctagtt aagaacccaa cccacctaaa
     6721 tggtattata atcaccagca acatgtttgg tgatatcatc tccgatgaag cctccgttat
     6781 cccaggttcc ttgggtttgt tgccatctgc gtccttggcc tctttgccag acaagaacac
     6841 cgcatttggt ttgtacgaac catgccacgg ttctgctcca gatttgccaa agaataaggt
     6901 caaccctatc gccactatct tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc
     6961 tgaagaaggt aaggccattg aagatgcagt taaaaaggtt ttggatgcag gtatcagaac
     7021 tggtgattta ggtggttcca acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt
     7081 taagaaaatc cttgcttaaa aagattctct ttttttatga tatttgtaca taaactttat
     7141 aaatgaaatt cataatagaa acgacacgaa attacaaaat ggaatatgtt catagggtag
     7201 acgaaactat atacgcaatc tacatacatt tatcaagaag gagaaaaagg aggatgtaaa
     7261 ggaatacagg taagcaaatt gatactaatg gctcaacgtg ataaggaaaa agaattgcac
     7321 tttaacatta atattgacaa ggaggagggc accacacaaa aagttaggtg taacagaaaa
     7381 tcatgaaact atgattccta atttatatat tggaggattt tctct
//
