LOCUS       sequence-438155-        7443 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 7443)
  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 7443)
  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-CFAPoptim
FEATURES             Location/Qualifiers
     source          1..7443
                     /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             4336..4365
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4366..4395
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4408..4655
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4424..4442)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4673..4692)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4674..4692)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4702..4719)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4702..4718)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4711..4733)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4859..5314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4946..4965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5156..5177
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5615..6022
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6023..7117
                     /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 agtggatccc ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacgcgggt
     3601 gcgggtgcgg gtatgcaagc tgttgttact caagaaccat ctgttactgt ttctccaggt
     3661 ggtactgtta tcttgacttg tggttctggt actggtgctg ttacttctgg tcactacgct
     3721 aactggttcc aacaaaagcc aggtcaagct ccaagagctt tgatcttcga cactgacaag
     3781 aagtactctt ggactccagg tagattctct ggttctttgt tgggtgctaa ggctgctttg
     3841 actatctctg acgctcaacc agaagacgaa gctgaatact actgttcttt gtctgacgtt
     3901 gacggttact tgttcggtgg tggtactcaa ttgactgttt tgtctggtgg tggtggttct
     3961 ggtggtggtg gttctggtgg tggtggttct ggtggtggtg gttctcaagc tgttgttact
     4021 caagaaccat ctgttactgt ttctccaggt ggtactgtta tcttgacttg tggttctggt
     4081 actggtgctg ttacttctgg tcactacgct aactggttcc aacaaaagcc aggtcaagct
     4141 ccaagagctt tgatcttcga cactgacaag aagtactctt ggactccagg tagattctct
     4201 ggttctttgt tgggtgctaa ggctgctttg actatctctg acgctcaacc agaagacgaa
     4261 gctgaatact actgttcttt gtctgacgtt gacggttact tgttcggtgg tggtactcaa
     4321 ttgactgttt tgtctgagca gaagctgatc agcgaggagg acctggagca gaagctgatc
     4381 agcgaggagg acctgtaacg cctcgagtca tgtaattagt tatgtcacgc ttacattcac
     4441 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     4501 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     4561 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     4621 agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag
     4681 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4741 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4801 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     4861 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4921 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4981 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5041 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5101 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5161 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5221 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5281 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     5341 atctgtgcgg tatttcacac cgcatatcga ccggtcgagg agaacttcta gtatatctac
     5401 atacctaata ttattgcctt attaaaaatg gaatcccaac aattacatca aaatccacat
     5461 tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa acgttatatt
     5521 tataggataa ttatactcta tttctcaaca agtaattggt tgtttggccg agcggtctaa
     5581 ggcgcctgat tcaagaaata tcttgaccgc agttaactgt gggaatactc aggtatcgta
     5641 agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca
     5701 cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag
     5761 aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag
     5821 agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc
     5881 acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta
     5941 gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat
     6001 atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt tgccaggtga
     6061 ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt
     6121 tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc
     6181 tacaggtgtt ccacttccag atgaggcgct ggaagcctcc aagaaggctg atgccgtttt
     6241 gttaggtgct gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt
     6301 actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc
     6361 cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt
     6421 tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga
     6481 tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat
     6541 ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct tggataaagc
     6601 taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga
     6661 attccctaca ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa
     6721 gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc
     6781 cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc
     6841 tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga
     6901 tttgccaaag aataaggtca accctatcgc cactatcttg tctgctgcaa tgatgttgaa
     6961 attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt
     7021 ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga
     7081 tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata
     7141 tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg
     7201 aatatgttca tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga
     7261 gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat
     7321 aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa
     7381 gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc
     7441 tct
//
