LOCUS       sequence-429708-        6019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synchronize trafficking of GPI-AP from the ER (RUCH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6019)
  AUTHORS   Bottone S, Joliot O, Cakil ZV, El Hajji L, Rakotoarison LM, 
            Boncompain G, Perez F, Gautier A
  TITLE     A fluorogenic chemically induced dimerization technology for 
            controlling, imaging and sensing protein proximity.
  JOURNAL   Nat Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8.
  PUBMED    37640938
REFERENCE   2  (bases 1 to 6019)
  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: "Nat 
            Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8."
COMMENT     Sequence Label: pFP5290 ss-FIREmate-HA- KDEL-IVS-IRES-
            ss-SBP-...
FEATURES             Location/Qualifiers
     source          1..6019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1431..1457
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          1550..1779
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1762..1781
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1851..2425,2426..2428,2429..2437)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2018..2035)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2245..2264
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     2451..2510
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             2511..2621
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             2631..3335
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAF
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNFE
                     DGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALKS
                     EIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     3041..3060
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3639..3760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3676..3695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3862..3878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3862..3878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3875..3897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3886..3902
                     /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)."
     promoter        complement(join(3910..3916,3917..3934,3935..3940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3955..3976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4093..4110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4264..4852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4344..4363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5023..5814,5815..5883))
                     /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     5646..5665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5884..5988)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
      961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgagc
     1081 atgttgcctt tggcagtgag gacatcgaga acactctggc caatatggac gacgaacaac
     1141 tggataggtt ggcctttggc gtaattcagc tcgatggtga cgggaatatc ctgctgtaca
     1201 atgctgctga aggggacatc actggcagag atcccaaaca ggtgattggg aagaacttct
     1261 tcaaggatgt tgcacctgga acggatactc ccgagtttta cggcaaattc aaggaaggcg
     1321 cagcgtcagg gaatctgaac accatgttcg aatggacgat accgacaagc aggggaccaa
     1381 ccaaggtcaa ggtgcacttg aagaaagccc tttccggagg tggcggaagt tatccttatg
     1441 acgtaccaga ctacgcactc ggaccgaaag atgaactgtg agcggccgca tagataactg
     1501 atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg tgagtactcc
     1561 ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat
     1621 ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa
     1681 aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg gccatacact
     1741 tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac
     1801 tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc cccctctccc
     1861 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1921 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1981 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     2041 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     2101 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     2161 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     2221 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     2281 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2341 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2401 ttcctttgaa aaacacgatg ataagcttgc cacaacccgg gaggcgcgcc atgtacagga
     2461 tgcaactcct gtcttgcatt gcactaagtc ttgcacttgt cacgaattcc gacgagaaga
     2521 ccactggttg gcgaggtgga cacgttgttg aaggactggc tggggaactt gaacaacttc
     2581 gtgcacgact ggagcatcac ccacaaggtc aacgtgaacc acctgcaggt gtgagcaagg
     2641 gcgaggagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     2701 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2761 cctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2821 tcctgtcccc tcagttcatg tacggctcca aggtctacat taagcaccca gccgacatcc
     2881 ccgactactt caagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg
     2941 aggacggcgg cattattcac gttaaccagg actcctccct gcaggacggc gtgttcatct
     3001 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     3061 ccatgggctg ggaggcctcc gaggagcgga tgtaccccga ggacggcgcc ctgaagagcg
     3121 agatcaagaa gaggctgaag ctgaaggacg gcggccacta cgccgccgag gtcaagacca
     3181 cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcgac atcaagttgg
     3241 acatcgtgtc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     3301 gccactccac cggcggcatg gacgagctgt acaagggccg gccaggcggc ggctccggtg
     3361 acagatattg ggtctttgtg aaacgggtgg gaggtggatc acgtacgctg gaaaatggcg
     3421 gaacctctct gtctgaaaaa actgtgctgc tgctggtgac tccctttctg gccgctgctt
     3481 ggtccctcca cccatgaggc ctcgagggcc aattaattaa ctctagataa ctgatcataa
     3541 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3601 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3661 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3721 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtcgag tgcattctag
     3781 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3841 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6001 aaaagtgcca cctgacgtc
//
