LOCUS       sequence-438042-        7278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP insert for C-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 7278)
  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 7278)
  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-CUP1pr-CFAPoptim
FEATURES             Location/Qualifiers
     source          1..7278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(167..186)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     286..308
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(346..364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    406..909
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        946..1050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1051..1119,1120..1911)
                     /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(1269..1288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2082..2670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2571..2590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2824..2841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2958..2979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2994..2999,3000..3017,3018..3024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3032..3048
                     /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     3037..3059
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3056..3072
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3056..3072
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3091..3111
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3093..3111
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3130..3364
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     primer_bind     complement(3410..3426)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3411..3427)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             3450..4169
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     CDS             4170..4199
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4200..4229
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4242..4489
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4258..4276)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4507..4526)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4508..4526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4536..4553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4536..4552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4545..4567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4693..5148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4780..4799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4990..5011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5449..5856
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5857..6951
                     /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 aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta
       61 agtcaatacc ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc
      121 tgtcagaaac ggccttaacg acgtagtcga tatggtgcac tctcagtaca atctgctctg
      181 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      241 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      301 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
      361 tatttttata ggttaatgtc atgataataa tggtttctta ggacggatcg cttgcctgta
      421 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
      481 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac
      541 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac
      601 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag
      661 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat
      721 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc
      781 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt
      841 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata
      901 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      961 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1021 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1081 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1141 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1201 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1261 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1321 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1381 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1441 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1501 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1561 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1621 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1681 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1741 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1801 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1861 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1921 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1981 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2041 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2101 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2161 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2221 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2281 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2341 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2401 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2461 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2521 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2581 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2641 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2701 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2761 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2821 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2881 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2941 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     3001 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     3061 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     3121 ctggagctcc tagttagaaa aagacatttt tgctgtcagt cactgtcaag agattctttt
     3181 gctggcattt cttctagaag caaaaagagc gatgcgtctt ttccgctgaa ccgttccagc
     3241 aaaaaagact accaacgcaa tatggattgt cagaatcata taaaagagaa gcaaataact
     3301 ccttgtcttg tatcaattgc attataatat cttcttgtta gtgcaatatc atatagaagt
     3361 catcgactag tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg
     3421 acctcgacgc gggtgcgggt gcgggtatgc aggccgtcgt tacccaagaa cctagtgtta
     3481 ccgttagccc aggtggtact gttatactta cttgtggaag tggtacgggt gccgtcacat
     3541 ctggtcatta tgcaaattgg tttcaacaaa aaccaggaca agctccaaga gctttgattt
     3601 ttgatactga taagaagtat tcttggaccc caggtagatt ttctggatct ttgctgggag
     3661 caaaggcagc tttgacaata tcagatgctc agcctgagga cgaagccgag tattactgtt
     3721 ctcttagcga cgtggatggc tacttgtttg gcggtggaac acaactgacg gttctgtccg
     3781 gtggtggcgg ctctggtggc ggtggcagcg gcggtggtgg ttccggaggc ggcggttctc
     3841 aggctgtggt gactcaggag ccgtcagtga ctgtgtcccc aggagggaca gtcattctca
     3901 cttgtggctc cggcactgga gctgtcacca gtggtcatta tgccaactgg ttccagcaga
     3961 agcctggcca agcccccagg gcacttatat ttgacaccga caagaagtat tcctggaccc
     4021 ctggccgatt ctcaggctcc ctccttgggg ccaaggctgc cctgaccatc tcggatgcgc
     4081 agcctgaaga tgaggctgag tattactgtt cgctctccga cgttgacggt tatctgttcg
     4141 gaggaggcac ccagctgacc gtcctctccg agcagaagct gatcagcgag gaggacctgg
     4201 agcagaagct gatcagcgag gaggacctgt aacgcctcga gtcatgtaat tagttatgtc
     4261 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     4321 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     4381 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     4441 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc
     4501 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     4561 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4621 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4681 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4741 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4801 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4861 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4921 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4981 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5041 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5101 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     5161 ttttctcctt acgcatctgt gcggtatttc acaccgcata tcgaccggtc gaggagaact
     5221 tctagtatat ctacatacct aatattattg ccttattaaa aatggaatcc caacaattac
     5281 atcaaaatcc acattctctt caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc
     5341 aaaaacgtta tatttatagg ataattatac tctatttctc aacaagtaat tggttgtttg
     5401 gccgagcggt ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa ctgtgggaat
     5461 actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt tttttcctca
     5521 acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga ctggaaattt
     5581 tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa attgggagaa
     5641 aaaggaaagg tgagagcgcc ggaaccggct tttcatatag aatagagaag cgttcatgac
     5701 taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt
     5761 ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca
     5821 atatatatat atatatttca aggatatacc attctaatgt ctgcccctaa gaagatcgtc
     5881 gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa
     5941 gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt
     6001 gctgctatcg atgctacagg tgttccactt ccagatgagg cgctggaagc ctccaagaag
     6061 gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga
     6121 cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca
     6181 tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa
     6241 ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag
     6301 gaagacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa
     6361 agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg
     6421 tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa
     6481 accatcaaga acgaattccc tacattgaag gttcaacatc aattgattga ttctgccgcc
     6541 atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt
     6601 ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct
     6661 gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac
     6721 ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat cttgtctgct
     6781 gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca
     6841 gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc
     6901 accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta aaaagattct
     6961 ctttttttat gatatttgta cataaacttt ataaatgaaa ttcataatag aaacgacacg
     7021 aaattacaaa atggaatatg ttcatagggt agacgaaact atatacgcaa tctacataca
     7081 tttatcaaga aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa ttgatactaa
     7141 tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac aaggaggagg
     7201 gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc taatttatat
     7261 attggaggat tttctcta
//
