LOCUS       sequence-438160-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP-tagged marker for the nucleus (FAP optimized for yeast 
            expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  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 6858)
  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-TEF1pr-RPA34-FAPoptim
FEATURES             Location/Qualifiers
     source          1..6858
                     /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        3162..3560
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3564..3580
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3564..3580
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5067..5096
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5097..5126
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5161..5180)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5162..5180)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5190..5207)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5190..5206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5199..5221)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5347..5802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5434..5453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5644..5665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6062..6724)
                     /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(6725..6858,1..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 tcatagcttc aaaatgtttc
     3181 tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa
     3241 acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac
     3301 ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa
     3361 aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt
     3421 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     3481 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     3541 gcatagcaat ctaatctaag ttttctagaa ctagtggatc catgtccaag ctttcgaaag
     3601 attacgtatc agattcagac tctgatgatg aagtgatatc aaacgagttc agcataccag
     3661 atggtttcaa gaaatgtaaa catttaaaga atttccctct caacggtgat aacaaaaaaa
     3721 aagcgaaaca gcaacaggtt tggctgatca aattcccatc aaatgtggac atctccaagt
     3781 taaaatctct gcctgtagat tttgaatcct ctacaaccat gacaattgat aaacacgatt
     3841 ataaaatcat ggacgataca gatattgagt cctcgttaac acaggataac ctcagtaata
     3901 tgactttatt ggttccttcg gaaagtaagg aaagtcttaa aattgctagc actgccaagg
     3961 ataatgcacc attacagttt gacaaggtat tttctgtcag cgaaaccgct aaaatcccag
     4021 ctattgatta cagtaaggta agagttccac gtaaagatgt tccaaaagta gaaggcttga
     4081 agttggagca ttttgctact ggttatgatg ccgaagattt ccatgtggct gaggaagtta
     4141 aagaaaataa gaaagaacca aagaagcgtt cacatcacga cgatgaagag gagtctagtg
     4201 aaaagaagaa aaagaagaag gagaagaggg agaagaggga gaagaaggat aaaaaggata
     4261 agaaaaagaa acatagagat cccgggctgc aggaattcga tatcaagctt atcgataccg
     4321 tcgacgcggg tgcgggtgcg ggtatgcaag ctgttgttac tcaagaacca tctgttactg
     4381 tttctccagg tggtactgtt atcttgactt gtggttctgg tactggtgct gttacttctg
     4441 gtcactacgc taactggttc caacaaaagc caggtcaagc tccaagagct ttgatcttcg
     4501 acactgacaa gaagtactct tggactccag gtagattctc tggttctttg ttgggtgcta
     4561 aggctgcttt gactatctct gacgctcaac cagaagacga agctgaatac tactgttctt
     4621 tgtctgacgt tgacggttac ttgttcggtg gtggtactca attgactgtt ttgtctggtg
     4681 gtggtggttc tggtggtggt ggttctggtg gtggtggttc tggtggtggt ggttctcaag
     4741 ctgttgttac tcaagaacca tctgttactg tttctccagg tggtactgtt atcttgactt
     4801 gtggttctgg tactggtgct gttacttctg gtcactacgc taactggttc caacaaaagc
     4861 caggtcaagc tccaagagct ttgatcttcg acactgacaa gaagtactct tggactccag
     4921 gtagattctc tggttctttg ttgggtgcta aggctgcttt gactatctct gacgctcaac
     4981 cagaagacga agctgaatac tactgttctt tgtctgacgt tgacggttac ttgttcggtg
     5041 gtggtactca attgactgtt ttgtctgagc agaagctgat cagcgaggag gacctggagc
     5101 agaagctgat cagcgaggag gacctgtaac gcctcgaggg ggggcccggt acccaattcg
     5161 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5221 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5281 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5341 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5401 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5461 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5521 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5581 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5641 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5701 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5761 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     5821 ccttacgcat ctgtgcggta tttcacaccg catagatccg tcgagttcaa gagaaaaaaa
     5881 aagaaaaagc aaaaagaaaa aaggaaagcg cgcctcgttc agaatgacac gtatagaatg
     5941 atgcattacc ttgtcatctt cagtatcata ctgttcgtat acatacttac tgacattcat
     6001 aggtatacat atatacacat gtatatatat cgtatgctgc agctttaaat aatcggtgtc
     6061 actacataag aacacctttg gtggagggaa catcgttggt accattgggc gaggtggctt
     6121 ctcttatggc aaccgcaaga gccttgaacg cactctcact acggtgatga tcattcttgc
     6181 ctcgcagaca atcaacgtgg agggtaattc tgctagcctc tgcaaagctt tcaagaaaat
     6241 gcgggatcat ctcgcaagag agatctccta ctttctccct ttgcaaacca agttcgacaa
     6301 ctgcgtacgg cctgttcgaa agatctacca ccgctctgga aagtgcctca tccaaaggcg
     6361 caaatcctga tccaaacctt tttactccac gcacggcccc tagggcctct ttaaaagctt
     6421 gaccgagagc aatcccgcag tcttcagtgg tgtgatggtc gtctatgtgt aagtcaccaa
     6481 tgcactcaac gattagcgac cagccggaat gcttggccag agcatgtatc atatggtcca
     6541 gaaaccctat acctgtgtgg acgttaatca cttgcgattg tgtggcctgt tctgctactg
     6601 cttctgcctc tttttctggg aagatcgagt gctctatcgc taggggacca ccctttaaag
     6661 agatcgcaat ctgaatcttg gtttcatttg taatacgctt tactagggct ttctgctctg
     6721 tcatctttgc cttcgtttat cttgcctgct cattttttag tatattcttc gaagaaatca
     6781 cattacttta tataatgtat aattcattat gtgataatgc caatcgctaa gaaaaaaaaa
     6841 gagtcatccg ctaggtgg
//
