LOCUS       sequence-438032-        7443 bp    DNA     circular SYN 15-APR-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 7443)
  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 7443)
  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-TEF1pr-CFAPorig
FEATURES             Location/Qualifiers
     source          1..7443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(167..186)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     286..308
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(346..364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    406..909
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        946..1050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1051..1119,1120..1911)
                     /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(1269..1288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2082..2670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2571..2590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2824..2841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2958..2979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2994..2999,3000..3017,3018..3024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3032..3048
                     /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     3037..3059
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3056..3072
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3056..3072
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3091..3111
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3093..3111
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3129..3527
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3531..3547
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3531..3547
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3615..4334
                     /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             4335..4364
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4365..4394
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4407..4654
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4423..4441)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4672..4691)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4673..4691)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4701..4718)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4701..4717)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4710..4732)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4858..5313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4945..4964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5155..5176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5614..6021
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6022..7116
                     /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 aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta
       61 agtcaatacc ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc
      121 tgtcagaaac ggccttaacg acgtagtcga tatggtgcac tctcagtaca atctgctctg
      181 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      241 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      301 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
      361 tatttttata ggttaatgtc atgataataa tggtttctta ggacggatcg cttgcctgta
      421 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
      481 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac
      541 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac
      601 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag
      661 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat
      721 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc
      781 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt
      841 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata
      901 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      961 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1021 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1081 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1141 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1201 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1261 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1321 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1381 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1441 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1501 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1561 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1621 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1681 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1741 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1801 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1861 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1921 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1981 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2041 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2101 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2161 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2221 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2281 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2341 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2401 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2461 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2521 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2581 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2641 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2701 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2761 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2821 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2881 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2941 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     3001 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     3061 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     3121 ctggagctca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
     3181 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
     3241 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
     3301 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
     3361 cttgaaaatt ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt
     3421 aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt
     3481 ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt tctagaacta
     3541 gtggatcccc cgggctgcag gaattcgata tcaagcttat cgataccgtc gacgcgggtg
     3601 cgggtgcggg tatgcaggcc gtcgttaccc aagaacctag tgttaccgtt agcccaggtg
     3661 gtactgttat acttacttgt ggaagtggta cgggtgccgt cacatctggt cattatgcaa
     3721 attggtttca acaaaaacca ggacaagctc caagagcttt gatttttgat actgataaga
     3781 agtattcttg gaccccaggt agattttctg gatctttgct gggagcaaag gcagctttga
     3841 caatatcaga tgctcagcct gaggacgaag ccgagtatta ctgttctctt agcgacgtgg
     3901 atggctactt gtttggcggt ggaacacaac tgacggttct gtccggtggt ggcggctctg
     3961 gtggcggtgg cagcggcggt ggtggttccg gaggcggcgg ttctcaggct gtggtgactc
     4021 aggagccgtc agtgactgtg tccccaggag ggacagtcat tctcacttgt ggctccggca
     4081 ctggagctgt caccagtggt cattatgcca actggttcca gcagaagcct ggccaagccc
     4141 ccagggcact tatatttgac accgacaaga agtattcctg gacccctggc cgattctcag
     4201 gctccctcct tggggccaag gctgccctga ccatctcgga tgcgcagcct gaagatgagg
     4261 ctgagtatta ctgttcgctc tccgacgttg acggttatct gttcggagga ggcacccagc
     4321 tgaccgtcct ctccgagcag aagctgatca gcgaggagga cctggagcag aagctgatca
     4381 gcgaggagga cctgtaacgc ctcgagtcat gtaattagtt atgtcacgct tacattcacg
     4441 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     4501 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     4561 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     4621 gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt
     4681 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4741 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4801 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     4861 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4921 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4981 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5041 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5101 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5161 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5221 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5281 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     5341 tctgtgcggt atttcacacc gcatatcgac cggtcgagga gaacttctag tatatctaca
     5401 tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa aatccacatt
     5461 ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt
     5521 ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga gcggtctaag
     5581 gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca ggtatcgtaa
     5641 gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata acgagaacac
     5701 acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga
     5761 ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga
     5821 gcgccggaac cggcttttca tatagaatag agaagcgttc atgactaaat gcttgcatca
     5881 caatacttga agttgacaat attatttaag gacctattgt tttttccaat aggtggttag
     5941 caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata tatatatata
     6001 tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac
     6061 cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt
     6121 cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct
     6181 acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga tgccgttttg
     6241 ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta
     6301 ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc
     6361 gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt
     6421 gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat
     6481 ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg
     6541 gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct
     6601 aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa
     6661 ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag
     6721 aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc
     6781 gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct
     6841 ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat
     6901 ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa
     6961 ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg
     7021 gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat
     7081 gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat
     7141 ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga
     7201 atatgttcat agggtagacg aaactatata cgcaatctac atacatttat caagaaggag
     7261 aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct caacgtgata
     7321 aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag
     7381 ttaggtgtaa cagaaaatca tgaaactatg attcctaatt tatatattgg aggattttct
     7441 cta
//
