LOCUS       sequence-466682-        7575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAP-tagged marker for the ER (FAP optimized for yeast expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7575)
  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 7575)
  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-SEC61-FAPoptim
FEATURES             Location/Qualifiers
     source          1..7575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1048..1446
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     1450..1466
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1450..1466
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3670..3699
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3700..3729
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3764..3783)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3765..3783)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3793..3810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3793..3809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3802..3824)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3950..4405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4037..4056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4247..4268
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4665..5327)
                     /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(5328..5515)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5662..5681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5781..5803
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5841..5859)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5907..6410
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6440..6544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6545..6613,6614..7405)
                     /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(6763..6782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcat agcttcaaaa tgtttctact ccttttttac
     1081 tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag
     1141 catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt
     1201 tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat
     1261 ttttatcacg tttctttttc ttgaaaattt tttttttgat ttttttctct ttcgatgacc
     1321 tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
     1381 cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa
     1441 tctaagtttt ctagaactag tggatccatg tcctccaacc gtgttctaga cttgtttaag
     1501 ccatttgaat ctttcctgcc ggaagtgatt gctccagaaa ggaaggttcc atacaaccag
     1561 aaacttatct ggacaggcgt ttctctactg atctttttga ttctgggcca gattccgctg
     1621 tacgggatcg tgtccagtga gacttccgac cctctgtact ggctacgtgc catgctggcc
     1681 tccaaccgtg gtactttact ggaattgggt gtttcgccca tcatcacttc atctatgatt
     1741 ttccaatttt tgcagggtac tcagctttta caaatcagac ctgagagcaa gcaggacaga
     1801 gagctgttcc aaattgctca aaaggtgtgc gctattattc tgatcttggg ccaagccctt
     1861 gtggtcgtca tgacaggtaa ctacggtgcc ccttcggacc tcggattgcc catctgtttg
     1921 ttgttaatct ttcaattgat gtttgcatcg ctgattgtga tgttattaga cgaattgcta
     1981 tctaagggtt acggcttggg ttccggtatt tctctgttca cggcaaccaa tattgccgaa
     2041 caaattttct ggagagcgtt tgctcctact acagtcaatt ccggtcgtgg taaggagttc
     2101 gaaggtgctg tgattgcctt tttccatctt ttggctgtca gaaaggacaa gaaaagagcc
     2161 cttgtcgagg ccttttaccg taccaatcta cctaatatgt tccaagtgtt gatgaccgtg
     2221 gccatcttcc tctttgtttt atatttacaa ggcttccgtt acgaattgcc catcaggtca
     2281 accaaagtga gaggtcaaat tggtatctac cccatcaaac tcttttatac ttccaacacc
     2341 ccaatcatgt tgcagagtgc attgacttct aacattttct tgatctctca aatccttttc
     2401 cagaaatacc caaccaatcc attgattcgt ttgatcggtg tttggggtat caggccgggc
     2461 acccagggcc ctcaaatggc cttgagcggg ttggcctact acatccaacc attaatgtct
     2521 ttatccgaag ctcttctgga ccctatcaag accatcgtct acatcacatt tgttcttggt
     2581 tcatgcgcag tattttccaa gacatggatc gaaatctccg gcacttcccc acgtgacatt
     2641 gccaaacaat tcaaagatca aggcatggtc attaacggta agagagaaac ctccatttac
     2701 agagaattga agaagatcat tccaactgct gctgctttcg gcggtgctac catcggtgct
     2761 ctttctgttg gctccgacct actaggtact ttaggttctg gggcatccat tttgatggct
     2821 actaccacca tctacggcta ctacgaagct gccgccaagg aaggtgggtt tactaagaac
     2881 ctcgttccag gattttctga tttgatgaag cttatcgata ccgtcgacgc gggtgcgggt
     2941 gcgggtatgc aagctgttgt tactcaagaa ccatctgtta ctgtttctcc aggtggtact
     3001 gttatcttga cttgtggttc tggtactggt gctgttactt ctggtcacta cgctaactgg
     3061 ttccaacaaa agccaggtca agctccaaga gctttgatct tcgacactga caagaagtac
     3121 tcttggactc caggtagatt ctctggttct ttgttgggtg ctaaggctgc tttgactatc
     3181 tctgacgctc aaccagaaga cgaagctgaa tactactgtt ctttgtctga cgttgacggt
     3241 tacttgttcg gtggtggtac tcaattgact gttttgtctg gtggtggtgg ttctggtggt
     3301 ggtggttctg gtggtggtgg ttctggtggt ggtggttctc aagctgttgt tactcaagaa
     3361 ccatctgtta ctgtttctcc aggtggtact gttatcttga cttgtggttc tggtactggt
     3421 gctgttactt ctggtcacta cgctaactgg ttccaacaaa agccaggtca agctccaaga
     3481 gctttgatct tcgacactga caagaagtac tcttggactc caggtagatt ctctggttct
     3541 ttgttgggtg ctaaggctgc tttgactatc tctgacgctc aaccagaaga cgaagctgaa
     3601 tactactgtt ctttgtctga cgttgacggt tacttgttcg gtggtggtac tcaattgact
     3661 gttttgtctg agcagaagct gatcagcgag gaggacctgg agcagaagct gatcagcgag
     3721 gaggacctgt aacgcctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta
     3781 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3841 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3901 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
     3961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4021 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4081 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4141 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4201 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4261 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4321 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4381 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     4441 gtatttcaca ccgcatagat ccgtcgagtt caagagaaaa aaaaagaaaa agcaaaaaga
     4501 aaaaaggaaa gcgcgcctcg ttcagaatga cacgtataga atgatgcatt accttgtcat
     4561 cttcagtatc atactgttcg tatacatact tactgacatt cataggtata catatataca
     4621 catgtatata tatcgtatgc tgcagcttta aataatcggt gtcactacat aagaacacct
     4681 ttggtggagg gaacatcgtt ggtaccattg ggcgaggtgg cttctcttat ggcaaccgca
     4741 agagccttga acgcactctc actacggtga tgatcattct tgcctcgcag acaatcaacg
     4801 tggagggtaa ttctgctagc ctctgcaaag ctttcaagaa aatgcgggat catctcgcaa
     4861 gagagatctc ctactttctc cctttgcaaa ccaagttcga caactgcgta cggcctgttc
     4921 gaaagatcta ccaccgctct ggaaagtgcc tcatccaaag gcgcaaatcc tgatccaaac
     4981 ctttttactc cacgcacggc ccctagggcc tctttaaaag cttgaccgag agcaatcccg
     5041 cagtcttcag tggtgtgatg gtcgtctatg tgtaagtcac caatgcactc aacgattagc
     5101 gaccagccgg aatgcttggc cagagcatgt atcatatggt ccagaaaccc tatacctgtg
     5161 tggacgttaa tcacttgcga ttgtgtggcc tgttctgcta ctgcttctgc ctctttttct
     5221 gggaagatcg agtgctctat cgctagggga ccacccttta aagagatcgc aatctgaatc
     5281 ttggtttcat ttgtaatacg ctttactagg gctttctgct ctgtcatctt tgccttcgtt
     5341 tatcttgcct gctcattttt tagtatattc ttcgaagaaa tcacattact ttatataatg
     5401 tataattcat tatgtgataa tgccaatcgc taagaaaaaa aaagagtcat ccgctaggtg
     5461 gaaaaaaaaa aatgaaaatc attaccgagg cataaaaaaa tatagagtgt actagaggag
     5521 gccaagagta atagaaaaag aaaattgcgg gaaaggactg tgttatgact tccctgacta
     5581 atgccgtgtt caaacgatac ctggcagtga ctcctagcgc tcaccaagct cttaaaacgg
     5641 gaatttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5701 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5761 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5821 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5881 aataatggtt tcttagggtc cttttcatca cgtgctataa aaataattat aatttaaatt
     5941 ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata
     6001 gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt
     6061 atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa
     6121 gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct
     6181 atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt
     6241 ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta
     6301 ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt
     6361 attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtcaggtgg
     6421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7561 gatcaaagga tcttc
//
