LOCUS       sequence-466718-        7695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP insert for N-terminally tagging genes of interest (optimized FAP
            for yeast expression + 2xMYC tag) + MFA1 signal sequence to help 
            target constructs to the ER .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7695)
  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 7695)
  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-MFA-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..7695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(287..1381)
                     /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"
     promoter        complement(1382..1789)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2090..2545)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2227..2248)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2439..2458
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2671..2693
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2685..2702
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2686..2702
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2712..2731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2712..2730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2749..2996)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2962..2980
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     2998..3014
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2999..3015
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3069..3098)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3099..3128)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(3858..4055,4056..4112))
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYLDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKR"
     promoter        complement(4128..4526)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(4544..4564)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4544..4562)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4583..4599)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4583..4599)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4596..4618)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4607..4623
                     /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)."
     promoter        complement(join(4631..4637,4638..4655,4656..4661))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4676..4697
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4814..4831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4985..5573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5065..5084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5744..6535,6536..6604))
                     /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     6367..6386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6605..6709)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6746..7249
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7291..7309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7347..7369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7469..7488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catgattttc tgttacacct aactttttgt gtggtgccct cctccttgtc aatattaatg
       61 ttaaagtgca attctttttc cttatcacgt tgagccatta gtatcaattt gcttacctgt
      121 attcctttac atcctccttt ttctccttct tgataaatgt atgtagattg cgtatatagt
      181 ttcgtctacc ctatgaacat attccatttt gtaatttcgt gtcgtttcta ttatgaattt
      241 catttataaa gtttatgtac aaatatcata aaaaaagaga atctttttaa gcaaggattt
      301 tcttaacttc ttcggcgaca gcatcaccga cttcggtggt actgttggaa ccacctaaat
      361 caccagttct gatacctgca tccaaaacct ttttaactgc atcttcaatg gccttacctt
      421 cttcaggcaa gttcaatgac aatttcaaca tcattgcagc agacaagata gtggcgatag
      481 ggttgacctt attctttggc aaatctggag cagaaccgtg gcatggttcg tacaaaccaa
      541 atgcggtgtt cttgtctggc aaagaggcca aggacgcaga tggcaacaaa cccaaggaac
      601 ctgggataac ggaggcttca tcggagatga tatcaccaaa catgttgctg gtgattataa
      661 taccatttag gtgggttggg ttcttaacta ggatcatggc ggcagaatca atcaattgat
      721 gttgaacctt caatgtaggg aattcgttct tgatggtttc ctccacagtt tttctccata
      781 atcttgaaga ggccaaaaca ttagctttat ccaaggacca aataggcaat ggtggctcat
      841 gttgtagggc catgaaagcg gccattcttg tgattctttg cacttctgga acggtgtatt
      901 gttcactatc ccaagcgaca ccatcaccat cgtcttcctt tctcttacca aagtaaatac
      961 ctcccactaa ttctctgaca acaacgaagt cagtaccttt agcaaattgt ggcttgattg
     1021 gagataagtc taaaagagag tcggatgcaa agttacatgg tcttaagttg gcgtacaatt
     1081 gaagttcttt acggattttt agtaaacctt gttcaggtct aacactaccg gtaccccatt
     1141 taggaccacc cacagcacct aacaaaacgg catcagcctt cttggaggct tccagcgcct
     1201 catctggaag tggaacacct gtagcatcga tagcagcacc accaattaaa tgattttcga
     1261 aatcgaactt gacattggaa cgaacatcag aaatagcttt aagaacctta atggcttcgg
     1321 ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac gatcttctta ggggcagaca
     1381 ttagaatggt atatccttga aatatatata tatatattgc tgaaatgtaa aaggtaagaa
     1441 aagttagaaa gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata
     1501 atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta
     1561 tatgaaaagc cggttccggc gctctcacct ttcctttttc tcccaatttt tcagttgaaa
     1621 aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa tttccagtca tcgaatttga
     1681 ttctgtgcga tagcgcccct gtgtgttctc gttatgttga ggaaaaaaat aatggttgct
     1741 aagagattcg aactcttgca tcttacgata cctgagtatt cccacagtta actgcggtca
     1801 agatatttct tgaatcaggc gccttagacc gctcggccaa acaaccaatt acttgttgag
     1861 aaatagagta taattatcct ataaatataa cgtttttgaa cacacatgaa caaggaagta
     1921 caggacaatt gattttgaag agaatgtgga ttttgatgta attgttggga ttccattttt
     1981 aataaggcaa taatattagg tatgtagata tactagaagt tctcctcgac cggtcgatat
     2041 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     2101 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2161 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2221 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2281 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2341 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2401 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2461 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2521 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     2581 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     2641 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     2701 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccggccgc aaattaaagc
     2761 cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac
     2821 gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat
     2881 aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt
     2941 tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt acatgactcg
     3001 aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggcg acccgcaccc
     3061 gcacccgcca ggtcctcctc gctgatcagc ttctgctcca ggtcctcctc gctgatcagc
     3121 ttctgctcag acaaaacagt caattgagta ccaccaccga acaagtaacc gtcaacgtca
     3181 gacaaagaac agtagtattc agcttcgtct tctggttgag cgtcagagat agtcaaagca
     3241 gccttagcac ccaacaaaga accagagaat ctacctggag tccaagagta cttcttgtca
     3301 gtgtcgaaga tcaaagctct tggagcttga cctggctttt gttggaacca gttagcgtag
     3361 tgaccagaag taacagcacc agtaccagaa ccacaagtca agataacagt accacctgga
     3421 gaaacagtaa cagatggttc ttgagtaaca acagcttgag aaccaccacc accagaacca
     3481 ccaccaccag aaccaccacc accagaacca ccaccaccag acaaaacagt caattgagta
     3541 ccaccaccga acaagtaacc gtcaacgtca gacaaagaac agtagtattc agcttcgtct
     3601 tctggttgag cgtcagagat agtcaaagca gccttagcac ccaacaaaga accagagaat
     3661 ctacctggag tccaagagta cttcttgtca gtgtcgaaga tcaaagctct tggagcttga
     3721 cctggctttt gttggaacca gttagcgtag tgaccagaag taacagcacc agtaccagaa
     3781 ccacaagtca agataacagt accacctgga gaaacagtaa cagatggttc ttgagtaaca
     3841 acagcttgca tggatcctct tttatccaaa gatacccctt cttctttagc agcaatgctg
     3901 gcaatagtag tatttataaa caataacccg ttatttgtgc tgttggaaaa tggcaaaaca
     3961 gcaacatcga aatccccttc taaatctaag taaccgatga cagcttcagc cggaatttgt
     4021 gccgtttcat cttctgttgt agtgttgact ggagcagcta atgcggagga tgctgcgaat
     4081 aaaactgcag taaaaattga aggaaatctc atactagttc tagaaaactt agattagatt
     4141 gctatgcttt ctttctaatg agcaagaagt aaaaaaagtt gtaatagaac aagaaaaatg
     4201 aaactgaaac ttgagaaatt gaagaccgtt tattaactta aatatcaatg ggaggtcatc
     4261 gaaagagaaa aaaatcaaaa aaaaaatttt caagaaaaag aaacgtgata aaaattttta
     4321 ttgccttttt cgacgaagaa aaagaaacga ggcggtctct tttttctttt ccaaaccttt
     4381 agtacgggta attaacgaca ccctagagga agaaagaggg gaaatttagt atgctgtgct
     4441 tgggtgtttt gaagtggtac ggcgatgcgc ggagtccgag aaaatctgga agagtaaaaa
     4501 aggagtagaa acattttgaa gctatgagct ccagcttttg ttccctttag tgagggttaa
     4561 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4621 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4681 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5401 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6721 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     6781 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     6841 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     6901 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     6961 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     7021 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     7081 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     7141 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     7201 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa
     7261 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7321 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7381 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     7441 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7501 catatcgact acgtcgttaa ggccgtttct gacagagtaa aattcttgag ggaactttca
     7561 ccattatggg aaatggttca agaaggtatt gacttaaact ccatcaaatg gtcaggtcat
     7621 tgagtgtttt ttatttgttg tatttttttt tttttagaga aaatcctcca atatataaat
     7681 taggaatcat agttt
//
