LOCUS       sequence-466719-        6159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP insert for C-terminally tagging genes of interest (original FAP 
            + 2xMYC tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  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 6159)
  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-CFAPorig
FEATURES             Location/Qualifiers
     source          1..6159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(57..244)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(391..410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     510..532
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(570..588)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    636..1139
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1169..1273
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1274..1342,1343..2134)
                     /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(1492..1511)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2305..2893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2794..2813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3047..3064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3181..3202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3217..3222,3223..3240,3241..3247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3255..3271
                     /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     3260..3282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3279..3295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3279..3295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3314..3334
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3316..3334
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3352..3750
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3754..3770
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3754..3770
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3838..4557
                     /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             4558..4587
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4588..4617
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4652..4671)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4653..4671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4681..4698)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4681..4697)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4690..4712)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4838..5293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4925..4944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5135..5156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5553..6159,1..56))
                     /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"
ORIGIN
        1 atctgaatct tggtttcatt tgtaatacgc tttactaggg ctttctgctc tgtcatcttt
       61 gccttcgttt atcttgcctg ctcatttttt agtatattct tcgaagaaat cacattactt
      121 tatataatgt ataattcatt atgtgataat gccaatcgct aagaaaaaaa aagagtcatc
      181 cgctaggtgg aaaaaaaaaa atgaaaatca ttaccgaggc ataaaaaaat atagagtgta
      241 ctagaggagg ccaagagtaa tagaaaaaga aaattgcggg aaaggactgt gttatgactt
      301 ccctgactaa tgccgtgttc aaacgatacc tggcagtgac tcctagcgct caccaagctc
      361 ttaaaacggg aatttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      421 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      481 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      541 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      601 tgtcatgata ataatggttt cttagggtcc ttttcatcac gtgctataaa aataattata
      661 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
      721 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
      781 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
      841 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
      901 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
      961 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     1021 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     1081 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     1141 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1201 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1261 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1321 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1381 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1441 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1501 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1561 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1621 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1681 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1741 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1801 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1861 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1921 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1981 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2041 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2101 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2161 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2761 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2821 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2881 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2941 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3001 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3061 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3121 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3181 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3241 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     3301 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tcatagcttc
     3361 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     3421 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     3481 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc
     3541 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     3601 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
     3661 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
     3721 tagaaagaaa gcatagcaat ctaatctaag ttttctagaa ctagtggatc ccccgggctg
     3781 caggaattcg atatcaagct tatcgatacc gtcgacgcgg gtgcgggtgc gggtatgcag
     3841 gccgtcgtta cccaagaacc tagtgttacc gttagcccag gtggtactgt tatacttact
     3901 tgtggaagtg gtacgggtgc cgtcacatct ggtcattatg caaattggtt tcaacaaaaa
     3961 ccaggacaag ctccaagagc tttgattttt gatactgata agaagtattc ttggacccca
     4021 ggtagatttt ctggatcttt gctgggagca aaggcagctt tgacaatatc agatgctcag
     4081 cctgaggacg aagccgagta ttactgttct cttagcgacg tggatggcta cttgtttggc
     4141 ggtggaacac aactgacggt tctgtccggt ggtggcggct ctggtggcgg tggcagcggc
     4201 ggtggtggtt ccggaggcgg cggttctcag gctgtggtga ctcaggagcc gtcagtgact
     4261 gtgtccccag gagggacagt cattctcact tgtggctccg gcactggagc tgtcaccagt
     4321 ggtcattatg ccaactggtt ccagcagaag cctggccaag cccccagggc acttatattt
     4381 gacaccgaca agaagtattc ctggacccct ggccgattct caggctccct ccttggggcc
     4441 aaggctgccc tgaccatctc ggatgcgcag cctgaagatg aggctgagta ttactgttcg
     4501 ctctccgacg ttgacggtta tctgttcgga ggaggcaccc agctgaccgt cctctccgag
     4561 cagaagctga tcagcgagga ggacctggag cagaagctga tcagcgagga ggacctgtaa
     4621 cgcctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     4681 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4741 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4801 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     4861 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4921 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4981 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5041 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5101 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5161 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5221 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5281 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5341 gcatagatcc gtcgagttca agagaaaaaa aaagaaaaag caaaaagaaa aaaggaaagc
     5401 gcgcctcgtt cagaatgaca cgtatagaat gatgcattac cttgtcatct tcagtatcat
     5461 actgttcgta tacatactta ctgacattca taggtataca tatatacaca tgtatatata
     5521 tcgtatgctg cagctttaaa taatcggtgt cactacataa gaacaccttt ggtggaggga
     5581 acatcgttgg taccattggg cgaggtggct tctcttatgg caaccgcaag agccttgaac
     5641 gcactctcac tacggtgatg atcattcttg cctcgcagac aatcaacgtg gagggtaatt
     5701 ctgctagcct ctgcaaagct ttcaagaaaa tgcgggatca tctcgcaaga gagatctcct
     5761 actttctccc tttgcaaacc aagttcgaca actgcgtacg gcctgttcga aagatctacc
     5821 accgctctgg aaagtgcctc atccaaaggc gcaaatcctg atccaaacct ttttactcca
     5881 cgcacggccc ctagggcctc tttaaaagct tgaccgagag caatcccgca gtcttcagtg
     5941 gtgtgatggt cgtctatgtg taagtcacca atgcactcaa cgattagcga ccagccggaa
     6001 tgcttggcca gagcatgtat catatggtcc agaaacccta tacctgtgtg gacgttaatc
     6061 acttgcgatt gtgtggcctg ttctgctact gcttctgcct ctttttctgg gaagatcgag
     6121 tgctctatcg ctaggggacc accctttaaa gagatcgca
//
