LOCUS       sequence-440010-        7269 bp    DNA     circular SYN 15-APR-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 7269)
  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 7269)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-CUP1pr-NFAPoptim
FEATURES             Location/Qualifiers
     source          1..7269
                     /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        3131..3365
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             4102..4131
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4132..4161
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4215..4231)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4216..4232)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4234..4481
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4250..4268)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4499..4518)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4500..4518)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4528..4545)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4528..4544)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4537..4559)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4685..5140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4772..4791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4982..5003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5441..5848
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5849..6943
                     /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 ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt
     3181 tgctggcatt tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag
     3241 caaaaaagac taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac
     3301 tccttgtctt gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag
     3361 tcatcgacta gtggatccat gcaagctgtt gttactcaag aaccatctgt tactgtttct
     3421 ccaggtggta ctgttatctt gacttgtggt tctggtactg gtgctgttac ttctggtcac
     3481 tacgctaact ggttccaaca aaagccaggt caagctccaa gagctttgat cttcgacact
     3541 gacaagaagt actcttggac tccaggtaga ttctctggtt ctttgttggg tgctaaggct
     3601 gctttgacta tctctgacgc tcaaccagaa gacgaagctg aatactactg ttctttgtct
     3661 gacgttgacg gttacttgtt cggtggtggt actcaattga ctgttttgtc tggtggtggt
     3721 ggttctggtg gtggtggttc tggtggtggt ggttctggtg gtggtggttc tcaagctgtt
     3781 gttactcaag aaccatctgt tactgtttct ccaggtggta ctgttatctt gacttgtggt
     3841 tctggtactg gtgctgttac ttctggtcac tacgctaact ggttccaaca aaagccaggt
     3901 caagctccaa gagctttgat cttcgacact gacaagaagt actcttggac tccaggtaga
     3961 ttctctggtt ctttgttggg tgctaaggct gctttgacta tctctgacgc tcaaccagaa
     4021 gacgaagctg aatactactg ttctttgtct gacgttgacg gttacttgtt cggtggtggt
     4081 actcaattga ctgttttgtc tgagcagaag ctgatcagcg aggaggacct ggagcagaag
     4141 ctgatcagcg aggaggacct ggcgggtgcg ggtgcgggtc gccccgggct gcaggaattc
     4201 gatatcaagc ttatcgatac cgtcgacctc gagtcatgta attagttatg tcacgcttac
     4261 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     4321 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     4381 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     4441 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     4501 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4561 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4621 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4681 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4741 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4801 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4861 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4921 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4981 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5041 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5101 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     5161 ttacgcatct gtgcggtatt tcacaccgca tatcgaccgg tcgaggagaa cttctagtat
     5221 atctacatac ctaatattat tgccttatta aaaatggaat cccaacaatt acatcaaaat
     5281 ccacattctc ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt tcaaaaacgt
     5341 tatatttata ggataattat actctatttc tcaacaagta attggttgtt tggccgagcg
     5401 gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt aactgtggga atactcaggt
     5461 atcgtaagat gcaagagttc gaatctctta gcaaccatta tttttttcct caacataacg
     5521 agaacacaca ggggcgctat cgcacagaat caaattcgat gactggaaat tttttgttaa
     5581 tttcagaggt cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa
     5641 ggtgagagcg ccggaaccgg cttttcatat agaatagaga agcgttcatg actaaatgct
     5701 tgcatcacaa tacttgaagt tgacaatatt atttaaggac ctattgtttt ttccaatagg
     5761 tggttagcaa tcgtcttact ttctaacttt tcttaccttt tacatttcag caatatatat
     5821 atatatattt caaggatata ccattctaat gtctgcccct aagaagatcg tcgttttgcc
     5881 aggtgaccac gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc
     5941 tgatgttcgt tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat
     6001 cgatgctaca ggtgttccac ttccagatga ggcgctggaa gcctccaaga aggctgatgc
     6061 cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta gacctgaaca
     6121 aggtttacta aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt
     6181 tgcatccgac tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga
     6241 cttcgttgtt gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagacga
     6301 tggtgatggt gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac
     6361 aagaatggcc gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga
     6421 taaagctaat gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa
     6481 gaacgaattc cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct
     6541 agttaagaac ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat
     6601 catctccgat gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt
     6661 ggcctctttg ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc
     6721 tccagatttg ccaaagaata aggtcaaccc tatcgccact atcttgtctg ctgcaatgat
     6781 gttgaaattg tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa
     6841 ggttttggat gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt
     6901 cggtgatgct gtcgccgaag aagttaagaa aatccttgct taaaaagatt ctcttttttt
     6961 atgatatttg tacataaact ttataaatga aattcataat agaaacgaca cgaaattaca
     7021 aaatggaata tgttcatagg gtagacgaaa ctatatacgc aatctacata catttatcaa
     7081 gaaggagaaa aaggaggatg taaaggaata caggtaagca aattgatact aatggctcaa
     7141 cgtgataagg aaaaagaatt gcactttaac attaatattg acaaggagga gggcaccaca
     7201 caaaaagtta ggtgtaacag aaaatcatga aactatgatt cctaatttat atattggagg
     7261 attttctct
//
