LOCUS       sequence-438027-        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP insert for C-terminally tagging genes of interest (optimized FAP
            for yeast expression + 2xMYC tag)+E2:E15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  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 7226)
  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-CFAPoptim
FEATURES             Location/Qualifiers
     source          1..7226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(200..219)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     319..341
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(379..397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    445..948
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        978..1082
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1083..1151,1152..1943)
                     /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(1301..1320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2114..2702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2603..2622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2856..2873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2990..3011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3026..3031,3032..3049,3050..3056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3064..3080
                     /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     3069..3091
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3088..3104
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3088..3104
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3123..3143
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3125..3143
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3912..4616
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     4630..4646
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4630..4646
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4714..5433
                     /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             5434..5463
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5464..5493
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5528..5547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5529..5547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5557..5574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5557..5573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5566..5588)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5714..6169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5801..5820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6011..6032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6429..7091)
                     /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(7092..7226,1..53))
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaaaaaaaaa tgaaaatcat taccgaggca taaaaaaata tagagtgtac tagaggaggc
       61 caagagtaat agaaaaagaa aattgcggga aaggactgtg ttatgacttc cctgactaat
      121 gccgtgttca aacgatacct ggcagtgact cctagcgctc accaagctct taaaacggga
      181 atttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
      241 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
      301 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
      361 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
      421 taatggtttc ttagggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt
      481 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
      541 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
      601 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga
      661 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
      721 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
      781 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
      841 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
      901 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tcaggtggca
      961 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1021 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1081 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1141 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1201 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1261 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1321 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1381 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1441 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1501 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1561 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1621 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1681 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1741 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1801 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1861 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1921 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1981 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2041 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2221 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2761 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2821 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2881 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2941 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3001 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     3061 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     3121 gcgcaattaa ccctcactaa agggaacaaa agctggagct cgccgggatc gaagaaatga
     3181 tggtaaatga aataggaaat caaggagcat gaaggcaaaa gacaaatata agggtcgaac
     3241 gaaaaataaa gtgaaaagtg ttgatatgat gtatttggct ttgcggcgcc gaaaaaacga
     3301 gtttacgcaa ttgcacaatc atgctgactc tgtggcggac ccgcgctctt gccggcccgg
     3361 cgataacgct gggcgtgagg ctgtgcccgg cggagttttt tgcgcctgca ttttccaagg
     3421 tttaccctgc gctaaggggc gagattggag aagcaataag aatgccggtt ggggttgcga
     3481 tgatgacgac cacgacaact ggtgtcatta tttaagttgc cgaaagaacc tgagtgcatt
     3541 tgcaacatga gtatactaga agaatgagcc aagacttgcg agacgcgagt ttgccggtgg
     3601 tgcgaacaat agagcgacca tgaccttgaa ggtgagacgc gcataaccgc tagagtactt
     3661 tgaagaggaa acagcaatag ggttgctacc agtataaata gacaggtaca tacaacactg
     3721 gaaatggttg tctgtttgag tacgctttca attcatttgg gtgtgcactt tattatgtta
     3781 caatatggaa gggaacttta cacttctcct atgcacatat attaattaaa gtccaatgct
     3841 agtagagaag gggggtaaca cccctccgcg ctcttttccg atttttttct aaaccgtgga
     3901 atatttcgga tatccttttg ttgtttccgg gtgtacaata tggacttcct cttttctggc
     3961 aaccaaaccc atacatcggg attcctataa taccttcgtt ggtctcccta acatgtaggt
     4021 ggcggagggg agatatacaa tagaacagat accagacaag acataatggg ctaaacaaga
     4081 ctacaccaat tacactgcct cattgatggt ggtacataac gaactaatac tgtagcccta
     4141 gacttgatag ccatcatcat atcgaagttt cactaccctt tttccatttg ccatctattg
     4201 aagtaataat aggcgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg
     4261 ttgtctcacc atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa
     4321 cgacaaagac agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc
     4381 acacgaaact ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac
     4441 ggccttcctt ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag
     4501 tataaataga cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc
     4561 ttccttgttt ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa
     4621 gctggccgct ctagaactag tggatccccc gggctgcagg aattcgatat caagcttatc
     4681 gataccgtcg acgcgggtgc gggtgcgggt atgcaggccg tcgttaccca agaacctagt
     4741 gttaccgtta gcccaggtgg tactgttata cttacttgtg gaagtggtac gggtgccgtc
     4801 acatctggtc attatgcaaa ttggtttcaa caaaaaccag gacaagctcc aagagctttg
     4861 atttttgata ctgataagaa gtattcttgg accccaggta gattttctgg atctttgctg
     4921 ggagcaaagg cagctttgac aatatcagat gctcagcctg aggacgaagc cgagtattac
     4981 tgttctctta gcgacgtgga tggctacttg tttggcggtg gaacacaact gacggttctg
     5041 tccggtggtg gcggctctgg tggcggtggc agcggcggtg gtggttccgg aggcggcggt
     5101 tctcaggctg tggtgactca ggagccgtca gtgactgtgt ccccaggagg gacagtcatt
     5161 ctcacttgtg gctccggcac tggagctgtc accagtggtc attatgccaa ctggttccag
     5221 cagaagcctg gccaagcccc cagggcactt atatttgaca ccgacaagaa gtattcctgg
     5281 acccctggcc gattctcagg ctccctcctt ggggccaagg ctgccctgac catctcggat
     5341 gcgcagcctg aagatgaggc tgagtattac tgttcgctct ccgacgttga cggttatctg
     5401 ttcggaggag gcacccagct gaccgtcctc tccgagcaga agctgatcag cgaggaggac
     5461 ctggagcaga agctgatcag cgaggaggac ctgtaacgcc tcgagggggg gcccggtacc
     5521 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     5581 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5641 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5701 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     6181 attttctcct tacgcatctg tgcggtattt cacaccgcat agatccgtcg agttcaagag
     6241 aaaaaaaaag aaaaagcaaa aagaaaaaag gaaagcgcgc ctcgttcaga atgacacgta
     6301 tagaatgatg cattaccttg tcatcttcag tatcatactg ttcgtataca tacttactga
     6361 cattcatagg tatacatata tacacatgta tatatatcgt atgctgcagc tttaaataat
     6421 cggtgtcact acataagaac acctttggtg gagggaacat cgttggtacc attgggcgag
     6481 gtggcttctc ttatggcaac cgcaagagcc ttgaacgcac tctcactacg gtgatgatca
     6541 ttcttgcctc gcagacaatc aacgtggagg gtaattctgc tagcctctgc aaagctttca
     6601 agaaaatgcg ggatcatctc gcaagagaga tctcctactt tctccctttg caaaccaagt
     6661 tcgacaactg cgtacggcct gttcgaaaga tctaccaccg ctctggaaag tgcctcatcc
     6721 aaaggcgcaa atcctgatcc aaaccttttt actccacgca cggcccctag ggcctcttta
     6781 aaagcttgac cgagagcaat cccgcagtct tcagtggtgt gatggtcgtc tatgtgtaag
     6841 tcaccaatgc actcaacgat tagcgaccag ccggaatgct tggccagagc atgtatcata
     6901 tggtccagaa accctatacc tgtgtggacg ttaatcactt gcgattgtgt ggcctgttct
     6961 gctactgctt ctgcctcttt ttctgggaag atcgagtgct ctatcgctag gggaccaccc
     7021 tttaaagaga tcgcaatctg aatcttggtt tcatttgtaa tacgctttac tagggctttc
     7081 tgctctgtca tctttgcctt cgtttatctt gcctgctcat tttttagtat attcttcgaa
     7141 gaaatcacat tactttatat aatgtataat tcattatgtg ataatgccaa tcgctaagaa
     7201 aaaaaaagag tcatccgcta ggtgga
//
