LOCUS       sequence-438156-        7752 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 7752)
  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 7752)
  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-MFA-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..7752
                     /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"
     sig_peptide     3787..3846
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4585..4614
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4615..4644
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4698..4714)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4699..4715)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4717..4964
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4733..4751)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4982..5001)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4983..5001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5011..5028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5011..5027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5020..5042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5168..5623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5255..5274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5465..5486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5924..6331
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6332..7426
                     /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 ttctagaatg
     3541 agatttcctt caatttttac tgcagtttta ttcgcagcat cctccgcatt agctgctcca
     3601 gtcaacacta caacagaaga tgaaacggca caaattccgg ctgaagctgt catcggttac
     3661 ttagatttag aaggggattt cgatgttgct gttttgccat tttccaacag cacaaataac
     3721 gggttattgt ttataaatac tactattgcc agcattgctg ctaaagaaga aggggtatct
     3781 ttggatatgg agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc
     3841 actggtgaca ctagtggatc catgcaagct gttgttactc aagaaccatc tgttactgtt
     3901 tctccaggtg gtactgttat cttgacttgt ggttctggta ctggtgctgt tacttctggt
     3961 cactacgcta actggttcca acaaaagcca ggtcaagctc caagagcttt gatcttcgac
     4021 actgacaaga agtactcttg gactccaggt agattctctg gttctttgtt gggtgctaag
     4081 gctgctttga ctatctctga cgctcaacca gaagacgaag ctgaatacta ctgttctttg
     4141 tctgacgttg acggttactt gttcggtggt ggtactcaat tgactgtttt gtctggtggt
     4201 ggtggttctg gtggtggtgg ttctggtggt ggtggttctg gtggtggtgg ttctcaagct
     4261 gttgttactc aagaaccatc tgttactgtt tctccaggtg gtactgttat cttgacttgt
     4321 ggttctggta ctggtgctgt tacttctggt cactacgcta actggttcca acaaaagcca
     4381 ggtcaagctc caagagcttt gatcttcgac actgacaaga agtactcttg gactccaggt
     4441 agattctctg gttctttgtt gggtgctaag gctgctttga ctatctctga cgctcaacca
     4501 gaagacgaag ctgaatacta ctgttctttg tctgacgttg acggttactt gttcggtggt
     4561 ggtactcaat tgactgtttt gtctgagcag aagctgatca gcgaggagga cctggagcag
     4621 aagctgatca gcgaggagga cctggcgggt gcgggtgcgg gtcgccccgg gctgcaggaa
     4681 ttcgatatca agcttatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct
     4741 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     4801 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     4861 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     4921 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc
     4981 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5041 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5101 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5161 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5221 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5281 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5341 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5401 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5461 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5521 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5581 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     5641 tccttacgca tctgtgcggt atttcacacc gcatatcgac cggtcgagga gaacttctag
     5701 tatatctaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa
     5761 aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa
     5821 cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga
     5881 gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca
     5941 ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata
     6001 acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt
     6061 taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg
     6121 aaaggtgaga gcgccggaac cggcttttca tatagaatag agaagcgttc atgactaaat
     6181 gcttgcatca caatacttga agttgacaat attatttaag gacctattgt tttttccaat
     6241 aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata
     6301 tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt
     6361 gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat
     6421 ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc
     6481 tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga
     6541 tgccgttttg ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga
     6601 acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa
     6661 ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac
     6721 tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga
     6781 cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat
     6841 cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt
     6901 ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat
     6961 caagaacgaa ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat
     7021 cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga
     7081 tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc
     7141 cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc
     7201 tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat
     7261 gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa
     7321 aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga
     7381 agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt
     7441 tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt
     7501 acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac atacatttat
     7561 caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct
     7621 caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc
     7681 acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt tatatattgg
     7741 aggattttct ct
//
