LOCUS       sequence-110505-        6686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synchronize trafficking of mCherry-GPI from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6686)
  AUTHORS   Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche 
            L, Mercanti V, Jollivet F, Raposo G, Perez F
  TITLE     Synchronization of secretory protein traffic in populations of 
            cells.
  JOURNAL   Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928.
  PUBMED    22406856
REFERENCE   2  (bases 1 to 6686)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-03-11- 11"; volume: "9"; 
            issue: "5"; pages: "493-8"
COMMENT     Sequence Label: ii-Str_SBP-mCherry-GPI
FEATURES             Location/Qualifiers
     source          1..6686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1753..1779
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2298..2527
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2510..2529
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2585..3159,3160..3162,3163..3171)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2752..2769)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2979..2998
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     3185..3244
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3245..3355
                     /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             3365..4069
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3507..3525)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3775..3794
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4310..4431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4347..4366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4533..4549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4533..4549)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4546..4568)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4557..4573
                     /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(4581..4587,4588..4605,4606..4611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4626..4647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4764..4781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4935..5523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5015..5034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5694..6485,6486..6554))
                     /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     6317..6336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6555..6659)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
      901 taccgagctc ggatcgatat ctgcggccta gctagcgctt aaggcctgtt aaccggtcgt
      961 acgtctccgg attcgaatta attcggatcc gtgatgcaca ggaggagagc cagggcctgt
     1021 cgggaagatc aaaagccagt catcgatgat cagcgcgacc ttatctccaa caatgagcaa
     1081 ctgcccatgc tgggccggcg ccctggggcc ccggagagca agtgcagccg cggagccctg
     1141 tacacaggct tttccatcct ggtgactctg ctcctcgctg gccaggccac caccgcctac
     1201 ttcctgtacc agcagcaggg ccggctggac aaactgacag tcacctccca gaacctgcag
     1261 ctcgagatgg accctagcaa agactcaaaa gctcaggtgt ccgctgccga ggctggcatt
     1321 actggaacat ggtacaatca gctcgggagc acctttattg tgactgctgg agccgatgga
     1381 gccctcaccg gaacatacga atctgctgtg ggaaacgccg aatcacggta cgtcctcact
     1441 ggccgatacg atagtgcccc tgccaccgac ggatctggga ctgccctggg atggactgtc
     1501 gcttggaaaa acaactaccg gaatgctcat tctgccacaa catggagtgg acagtacgtg
     1561 ggaggcgctg aggctagaat caatacacag tggctgctca catctggcac aaccgaggca
     1621 aatgcttgga aatccaccct ggtgggacat gacacattca ccaaagtgaa accctccgcc
     1681 gcttcaatcg atgccgccaa aaaagccgga gtcaacaacg gcaatcctct ggatgccgtc
     1741 cagcaggtcg actatccgta cgacgtacca gactacgcac tcgacctgca gctggagaac
     1801 ctgcgcatga agcttcccaa gcctcccaag cctgtgagca agatgcgcat ggccaccccg
     1861 ctgctgatgc atgcgctgcc catgggagcc ctgccccagg ggcccatgca gaatgccacc
     1921 aagtatggca acatgacaga ggaccatgtg atgcacctgc tccagaatgc tgaccccctg
     1981 aaggtgtacc cgccactgaa ggggagcttc ccggagaacc tgagacacct taagaacacc
     2041 atggagacca tagactggaa ggtctttgag agctggatgc accattggct cctgtttgaa
     2101 atgagcaggc actccttgga gcaaaagccc actgacgctc caccgaaaga gtcactggaa
     2161 ctggaggacc cgtcttctgg gctgggtgtg accaagcagg atctgggccc agtccccatg
     2221 tgaggatccg cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc
     2281 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag
     2341 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt
     2401 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg
     2461 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc
     2521 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat
     2581 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     2641 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     2701 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     2761 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     2821 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     2881 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     2941 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     3001 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     3061 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     3121 acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggaggcg
     3181 cgccatgtac aggatgcaac tcctgtcttg cattgcacta agtcttgcac ttgtcacgaa
     3241 ttccgacgag aagaccactg gttggcgagg tggacacgtt gttgaaggac tggctgggga
     3301 acttgaacaa cttcgtgcac gactggagca tcacccacaa ggtcaacgtg aaccacctgc
     3361 aggtgtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     3421 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3481 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     3541 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     3601 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     3661 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctacagga
     3721 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     3781 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     3841 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     3901 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     3961 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     4021 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg gccggccact
     4081 ggaaaatggc ggaacctctc tgtctgaaaa aactgtgctg ctgctggtga ctccctttct
     4141 ggccgctgct tggtccctcc acccatgagg cctcgagggc caattaatta actctagata
     4201 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4261 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtcga
     4441 gtgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4501 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4561 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4621 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4681 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4741 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4801 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4861 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5101 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5341 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6661 acatttcccc gaaaagtgcc acctga
//
