LOCUS       sequence-466689-        8469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP tagged SEC61 (C-terminally tagged, optimized) under the Tef1 
            promoter with 2xMYC tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8469)
  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 8469)
  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: pRS415-TEF1pr-SEC61-FAPoptim
FEATURES             Location/Qualifiers
     source          1..8469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(143..162)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      956..1544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1445..1464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1698..1715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1832..1853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1868..1873,1874..1891,1892..1898)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1906..1922
                     /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     1911..1933
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1930..1946
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1930..1946
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1965..1985
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1967..1985
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2003..2401
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2405..2421
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2405..2421
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4235..4264
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4265..4294
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4307..4554
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4323..4341)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4572..4591)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4573..4591)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4601..4618)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4601..4617)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4610..4632)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4758..5213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4845..4864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5055..5076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5514..5921
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5922..7016
                     /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"
     primer_bind     complement(7510..7529)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7629..7651
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7689..7707)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    7749..8252
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        8289..8393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8394..8462,8463..8469,1..785)
                     /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"
ORIGIN
        1 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
       61 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      121 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
      181 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
      241 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
      301 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
      361 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
      421 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
      481 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
      541 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
      601 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
      661 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
      721 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
      781 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
      841 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
      901 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      961 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1021 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1081 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1141 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1201 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1261 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1321 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1381 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1441 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1501 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1561 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1621 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1681 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     1741 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1801 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     1861 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     1921 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     1981 aaagggaaca aaagctggag ctcatagctt caaaatgttt ctactccttt tttactcttc
     2041 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     2101 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     2161 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     2221 tcacgtttct ttttcttgaa aatttttttt ttgatttttt tctctttcga tgacctccca
     2281 ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt
     2341 tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa
     2401 gttttctaga actagtggat ccatgtcctc caaccgtgtt ctagacttgt ttaagccatt
     2461 tgaatctttc ctgccggaag tgattgctcc agaaaggaag gttccataca accagaaact
     2521 tatctggaca ggcgtttctc tactgatctt tttgattctg ggccagattc cgctgtacgg
     2581 gatcgtgtcc agtgagactt ccgaccctct gtactggcta cgtgccatgc tggcctccaa
     2641 ccgtggtact ttactggaat tgggtgtttc gcccatcatc acttcatcta tgattttcca
     2701 atttttgcag ggtactcagc ttttacaaat cagacctgag agcaagcagg acagagagct
     2761 gttccaaatt gctcaaaagg tgtgcgctat tattctgatc ttgggccaag cccttgtggt
     2821 cgtcatgaca ggtaactacg gtgccccttc ggacctcgga ttgcccatct gtttgttgtt
     2881 aatctttcaa ttgatgtttg catcgctgat tgtgatgtta ttagacgaat tgctatctaa
     2941 gggttacggc ttgggttccg gtatttctct gttcacggca accaatattg ccgaacaaat
     3001 tttctggaga gcgtttgctc ctactacagt caattccggt cgtggtaagg agttcgaagg
     3061 tgctgtgatt gcctttttcc atcttttggc tgtcagaaag gacaagaaaa gagcccttgt
     3121 cgaggccttt taccgtacca atctacctaa tatgttccaa gtgttgatga ccgtggccat
     3181 cttcctcttt gttttatatt tacaaggctt ccgttacgaa ttgcccatca ggtcaaccaa
     3241 agtgagaggt caaattggta tctaccccat caaactcttt tatacttcca acaccccaat
     3301 catgttgcag agtgcattga cttctaacat tttcttgatc tctcaaatcc ttttccagaa
     3361 atacccaacc aatccattga ttcgtttgat cggtgtttgg ggtatcaggc cgggcaccca
     3421 gggccctcaa atggccttga gcgggttggc ctactacatc caaccattaa tgtctttatc
     3481 cgaagctctt ctggacccta tcaagaccat cgtctacatc acatttgttc ttggttcatg
     3541 cgcagtattt tccaagacat ggatcgaaat ctccggcact tccccacgtg acattgccaa
     3601 acaattcaaa gatcaaggca tggtcattaa cggtaagaga gaaacctcca tttacagaga
     3661 attgaagaag atcattccaa ctgctgctgc tttcggcggt gctaccatcg gtgctctttc
     3721 tgttggctcc gacctactag gtactttagg ttctggggca tccattttga tggctactac
     3781 caccatctac ggctactacg aagctgccgc caaggaaggt gggtttacta agaacctcgt
     3841 tccaggattt tctgatttga tgaagcttat cgataccgtc gacgcgggtg cgggtgcggg
     3901 tatgcaagct gttgttactc aagaaccatc tgttactgtt tctccaggtg gtactgttat
     3961 cttgacttgt ggttctggta ctggtgctgt tacttctggt cactacgcta actggttcca
     4021 acaaaagcca ggtcaagctc caagagcttt gatcttcgac actgacaaga agtactcttg
     4081 gactccaggt agattctctg gttctttgtt gggtgctaag gctgctttga ctatctctga
     4141 cgctcaacca gaagacgaag ctgaatacta ctgttctttg tctgacgttg acggttactt
     4201 gttcggtggt ggtactcaat tgactgtttt gtctgagcag aagctgatca gcgaggagga
     4261 cctggagcag aagctgatca gcgaggagga cctgtaacgc ctcgagtcat gtaattagtt
     4321 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     4381 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     4441 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
     4501 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg
     4561 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     4621 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4681 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4741 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4801 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4861 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4921 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4981 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5041 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5101 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5161 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     5221 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatcgac cggtcgagga
     5281 gaacttctag tatatctaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca
     5341 attacatcaa aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg
     5401 tgttcaaaaa cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt
     5461 gtttggccga gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg
     5521 ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt
     5581 cctcaacata acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga
     5641 aattttttgt taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg
     5701 gagaaaaagg aaaggtgaga gcgccggaac cggcttttca tatagaatag agaagcgttc
     5761 atgactaaat gcttgcatca caatacttga agttgacaat attatttaag gacctattgt
     5821 tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt
     5881 cagcaatata tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga
     5941 tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc
     6001 ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg
     6061 gtggtgctgc tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca
     6121 agaaggctga tgccgttttg ttaggtgctg tgggtggtcc taaatggggt accggtagtg
     6181 ttagacctga acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa
     6241 gaccatgtaa ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg
     6301 ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga
     6361 gaaaggaaga cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag
     6421 tgcaaagaat cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta
     6481 tttggtcctt ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg
     6541 aggaaaccat caagaacgaa ttccctacat tgaaggttca acatcaattg attgattctg
     6601 ccgccatgat cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca
     6661 tgtttggtga tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc
     6721 catctgcgtc cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat
     6781 gccacggttc tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt
     6841 ctgctgcaat gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag
     6901 atgcagttaa aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca
     6961 gtaccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag
     7021 attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg
     7081 acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac
     7141 atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat
     7201 actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga
     7261 ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt
     7321 tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg
     7381 acctgaccat ttgatggagt ttaagtcaat accttcttga accatttccc ataatggtga
     7441 aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtagt cgatatggtg
     7501 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     7561 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     7621 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     7681 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     7741 ttaggacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
     7801 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
     7861 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
     7921 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
     7981 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
     8041 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
     8101 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
     8161 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
     8221 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
     8281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     8341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     8401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     8461 ctcacccag
//
