LOCUS       sequence-438154-        7440 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 7440)
  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 7440)
  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-TEF1pr-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..7440
                     /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        3130..3528
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3532..3548
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3532..3548
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4273..4302
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4303..4332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4386..4402)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4387..4403)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4405..4652
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4421..4439)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4670..4689)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4671..4689)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4699..4716)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4699..4715)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4708..4730)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4856..5311
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4943..4962)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5153..5174
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5612..6019
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6020..7114
                     /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 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
     3181 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     3241 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     3301 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     3361 tcttgaaaat tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
     3421 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
     3481 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttctagaact
     3541 agtggatcca tgcaagctgt tgttactcaa gaaccatctg ttactgtttc tccaggtggt
     3601 actgttatct tgacttgtgg ttctggtact ggtgctgtta cttctggtca ctacgctaac
     3661 tggttccaac aaaagccagg tcaagctcca agagctttga tcttcgacac tgacaagaag
     3721 tactcttgga ctccaggtag attctctggt tctttgttgg gtgctaaggc tgctttgact
     3781 atctctgacg ctcaaccaga agacgaagct gaatactact gttctttgtc tgacgttgac
     3841 ggttacttgt tcggtggtgg tactcaattg actgttttgt ctggtggtgg tggttctggt
     3901 ggtggtggtt ctggtggtgg tggttctggt ggtggtggtt ctcaagctgt tgttactcaa
     3961 gaaccatctg ttactgtttc tccaggtggt actgttatct tgacttgtgg ttctggtact
     4021 ggtgctgtta cttctggtca ctacgctaac tggttccaac aaaagccagg tcaagctcca
     4081 agagctttga tcttcgacac tgacaagaag tactcttgga ctccaggtag attctctggt
     4141 tctttgttgg gtgctaaggc tgctttgact atctctgacg ctcaaccaga agacgaagct
     4201 gaatactact gttctttgtc tgacgttgac ggttacttgt tcggtggtgg tactcaattg
     4261 actgttttgt ctgagcagaa gctgatcagc gaggaggacc tggagcagaa gctgatcagc
     4321 gaggaggacc tggcgggtgc gggtgcgggt cgccccgggc tgcaggaatt cgatatcaag
     4381 cttatcgata ccgtcgacct cgagtcatgt aattagttat gtcacgctta cattcacgcc
     4441 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     4501 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     4561 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     4621 aggttttggg acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga
     4681 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4741 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4801 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     4861 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4921 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4981 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5041 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5101 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5161 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5221 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5281 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     5341 tgtgcggtat ttcacaccgc atatcgaccg gtcgaggaga acttctagta tatctacata
     5401 cctaatatta ttgccttatt aaaaatggaa tcccaacaat tacatcaaaa tccacattct
     5461 cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat
     5521 aggataatta tactctattt ctcaacaagt aattggttgt ttggccgagc ggtctaaggc
     5581 gcctgattca agaaatatct tgaccgcagt taactgtggg aatactcagg tatcgtaaga
     5641 tgcaagagtt cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac
     5701 aggggcgcta tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg
     5761 tcgcctgacg catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagc
     5821 gccggaaccg gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca
     5881 atacttgaag ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca
     5941 atcgtcttac tttctaactt ttcttacctt ttacatttca gcaatatata tatatatatt
     6001 tcaaggatat accattctaa tgtctgcccc taagaagatc gtcgttttgc caggtgacca
     6061 cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt ctgatgttcg
     6121 ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta tcgatgctac
     6181 aggtgttcca cttccagatg aggcgctgga agcctccaag aaggctgatg ccgttttgtt
     6241 aggtgctgtg ggtggtccta aatggggtac cggtagtgtt agacctgaac aaggtttact
     6301 aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact ttgcatccga
     6361 ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg acttcgttgt
     6421 tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaagacg atggtgatgg
     6481 tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca caagaatggc
     6541 cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg ataaagctaa
     6601 tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca agaacgaatt
     6661 ccctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc tagttaagaa
     6721 cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata tcatctccga
     6781 tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct tggcctcttt
     6841 gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg ctccagattt
     6901 gccaaagaat aaggtcaacc ctatcgccac tatcttgtct gctgcaatga tgttgaaatt
     6961 gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa aggttttgga
     7021 tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag tcggtgatgc
     7081 tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt tatgatattt
     7141 gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac aaaatggaat
     7201 atgttcatag ggtagacgaa actatatacg caatctacat acatttatca agaaggagaa
     7261 aaaggaggat gtaaaggaat acaggtaagc aaattgatac taatggctca acgtgataag
     7321 gaaaaagaat tgcactttaa cattaatatt gacaaggagg agggcaccac acaaaaagtt
     7381 aggtgtaaca gaaaatcatg aaactatgat tcctaattta tatattggag gattttctct
//
