LOCUS       sequence-438153-        7223 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 7223)
  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 7223)
  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-ADH1pr-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..7223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(201..220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     320..342
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(380..398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    446..949
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        979..1083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1084..1152,1153..1944)
                     /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(1302..1321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2115..2703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2604..2623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2857..2874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2991..3012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3027..3032,3033..3050,3051..3057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3065..3081
                     /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     3070..3092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3089..3105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3089..3105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3124..3144
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3126..3144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3913..4617
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     4631..4647
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4631..4647
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5372..5401
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5402..5431
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5485..5501)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5486..5502)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5526..5545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5527..5545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5555..5572)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5555..5571)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5564..5586)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5712..6167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5799..5818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6009..6030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6427..7089)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(join(7090..7223,1..54))
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaaaaaaaaa atgaaaatca ttaccgaggc ataaaaaaat atagagtgta ctagaggagg
       61 ccaagagtaa tagaaaaaga aaattgcggg aaaggactgt gttatgactt ccctgactaa
      121 tgccgtgttc aaacgatacc tggcagtgac tcctagcgct caccaagctc ttaaaacggg
      181 aatttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      241 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      301 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      361 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      421 ataatggttt cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt
      481 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
      541 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
      601 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
      661 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
      721 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
      781 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
      841 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
      901 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc
      961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2881 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3001 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3061 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3121 cgcgcaatta accctcacta aagggaacaa aagctggagc tcgccgggat cgaagaaatg
     3181 atggtaaatg aaataggaaa tcaaggagca tgaaggcaaa agacaaatat aagggtcgaa
     3241 cgaaaaataa agtgaaaagt gttgatatga tgtatttggc tttgcggcgc cgaaaaaacg
     3301 agtttacgca attgcacaat catgctgact ctgtggcgga cccgcgctct tgccggcccg
     3361 gcgataacgc tgggcgtgag gctgtgcccg gcggagtttt ttgcgcctgc attttccaag
     3421 gtttaccctg cgctaagggg cgagattgga gaagcaataa gaatgccggt tggggttgcg
     3481 atgatgacga ccacgacaac tggtgtcatt atttaagttg ccgaaagaac ctgagtgcat
     3541 ttgcaacatg agtatactag aagaatgagc caagacttgc gagacgcgag tttgccggtg
     3601 gtgcgaacaa tagagcgacc atgaccttga aggtgagacg cgcataaccg ctagagtact
     3661 ttgaagagga aacagcaata gggttgctac cagtataaat agacaggtac atacaacact
     3721 ggaaatggtt gtctgtttga gtacgctttc aattcatttg ggtgtgcact ttattatgtt
     3781 acaatatgga agggaacttt acacttctcc tatgcacata tattaattaa agtccaatgc
     3841 tagtagagaa ggggggtaac acccctccgc gctcttttcc gatttttttc taaaccgtgg
     3901 aatatttcgg atatcctttt gttgtttccg ggtgtacaat atggacttcc tcttttctgg
     3961 caaccaaacc catacatcgg gattcctata ataccttcgt tggtctccct aacatgtagg
     4021 tggcggaggg gagatataca atagaacaga taccagacaa gacataatgg gctaaacaag
     4081 actacaccaa ttacactgcc tcattgatgg tggtacataa cgaactaata ctgtagccct
     4141 agacttgata gccatcatca tatcgaagtt tcactaccct ttttccattt gccatctatt
     4201 gaagtaataa taggcgcatg caacttcttt tctttttttt tcttttctct ctcccccgtt
     4261 gttgtctcac catatccgca atgacaaaaa aatgatggaa gacactaaag gaaaaaatta
     4321 acgacaaaga cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatctcgaag
     4381 cacacgaaac tttttccttc cttcattcac gcacactact ctctaatgag caacggtata
     4441 cggccttcct tccagttact tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa
     4501 gtataaatag acctgcaatt attaatcttt tgtttcctcg tcattgttct cgttcccttt
     4561 cttccttgtt tctttttctg cacaatattt caagctatac caagcataca atcaactcca
     4621 agctggccgc tctagaacta gtggatccat gcaagctgtt gttactcaag aaccatctgt
     4681 tactgtttct ccaggtggta ctgttatctt gacttgtggt tctggtactg gtgctgttac
     4741 ttctggtcac tacgctaact ggttccaaca aaagccaggt caagctccaa gagctttgat
     4801 cttcgacact gacaagaagt actcttggac tccaggtaga ttctctggtt ctttgttggg
     4861 tgctaaggct gctttgacta tctctgacgc tcaaccagaa gacgaagctg aatactactg
     4921 ttctttgtct gacgttgacg gttacttgtt cggtggtggt actcaattga ctgttttgtc
     4981 tggtggtggt ggttctggtg gtggtggttc tggtggtggt ggttctggtg gtggtggttc
     5041 tcaagctgtt gttactcaag aaccatctgt tactgtttct ccaggtggta ctgttatctt
     5101 gacttgtggt tctggtactg gtgctgttac ttctggtcac tacgctaact ggttccaaca
     5161 aaagccaggt caagctccaa gagctttgat cttcgacact gacaagaagt actcttggac
     5221 tccaggtaga ttctctggtt ctttgttggg tgctaaggct gctttgacta tctctgacgc
     5281 tcaaccagaa gacgaagctg aatactactg ttctttgtct gacgttgacg gttacttgtt
     5341 cggtggtggt actcaattga ctgttttgtc tgagcagaag ctgatcagcg aggaggacct
     5401 ggagcagaag ctgatcagcg aggaggacct ggcgggtgcg ggtgcgggtc gccccgggct
     5461 gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc ccggtaccca
     5521 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     5581 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     5641 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     5701 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5761 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5821 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5881 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5941 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6001 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6061 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6121 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     6181 tttctcctta cgcatctgtg cggtatttca caccgcatag atccgtcgag ttcaagagaa
     6241 aaaaaaagaa aaagcaaaaa gaaaaaagga aagcgcgcct cgttcagaat gacacgtata
     6301 gaatgatgca ttaccttgtc atcttcagta tcatactgtt cgtatacata cttactgaca
     6361 ttcataggta tacatatata cacatgtata tatatcgtat gctgcagctt taaataatcg
     6421 gtgtcactac ataagaacac ctttggtgga gggaacatcg ttggtaccat tgggcgaggt
     6481 ggcttctctt atggcaaccg caagagcctt gaacgcactc tcactacggt gatgatcatt
     6541 cttgcctcgc agacaatcaa cgtggagggt aattctgcta gcctctgcaa agctttcaag
     6601 aaaatgcggg atcatctcgc aagagagatc tcctactttc tccctttgca aaccaagttc
     6661 gacaactgcg tacggcctgt tcgaaagatc taccaccgct ctggaaagtg cctcatccaa
     6721 aggcgcaaat cctgatccaa acctttttac tccacgcacg gcccctaggg cctctttaaa
     6781 agcttgaccg agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc
     6841 accaatgcac tcaacgatta gcgaccagcc ggaatgcttg gccagagcat gtatcatatg
     6901 gtccagaaac cctatacctg tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc
     6961 tactgcttct gcctcttttt ctgggaagat cgagtgctct atcgctaggg gaccaccctt
     7021 taaagagatc gcaatctgaa tcttggtttc atttgtaata cgctttacta gggctttctg
     7081 ctctgtcatc tttgccttcg tttatcttgc ctgctcattt tttagtatat tcttcgaaga
     7141 aatcacatta ctttatataa tgtataattc attatgtgat aatgccaatc gctaagaaaa
     7201 aaaaagagtc atccgctagg tgg
//
