LOCUS       sequence-110471-        7976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synchronize trafficking of ST from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  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 7976)
  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: Str-STIM1-NN_ST-SBP-mCherry
FEATURES             Location/Qualifiers
     source          1..7976
                     /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"
     intron          3587..3816
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3799..3818
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3874..4448,4449..4451,4452..4460)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4041..4058)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4268..4287
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4612..4725
                     /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="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             4735..5442
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4880..4898)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5148..5167
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5596..5717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5633..5652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5819..5835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5819..5835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5832..5854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5843..5859
                     /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(5867..5873,5874..5891,5892..5897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5912..5933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6050..6067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6221..6809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6301..6320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6980..7771,7772..7840))
                     /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     7603..7622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7841..7945)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ttaaacgacc
     1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
     1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
     1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
     1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
     1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
     1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
     1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
     1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
     1561 cgggcgccaa ctctgaggag tccactgcag cagagttttg ccgaattgac aagcccctgt
     1621 gtcacagtga ggatgagaaa ctcagcttcg aggcagtccg taacatccac aaactgatgg
     1681 acgatgatgc caatggtgat gtggatgtgg aagaaagtga tgagttcctg agggaagacc
     1741 tcaattacca tgacccaaca gtgaaacaca gcaccttcca tggtgaggat aagctcatca
     1801 gcgtggagga cctgtggaag gcatggaagt catcagaagt atacaattgg accgtggatg
     1861 aggtggtaca gtggctgatc acatatgtgg agctgcctca gtatgaggag accttccgga
     1921 agctgcagct cagtggccat gccatgccaa ggctggctgt caccaacacc accatgacag
     1981 ggactgtgct gaagatgaca gaccggagtc atcggcagaa gctgcagctg aaggctctgg
     2041 atacagtgct ctttgggcct cctctcttga ctcgccataa tcacctcaag gacttcatgc
     2101 tggtggtgtc tatcgttatt ggtgtgggcg gctgctggtt tgcctatatc cagaaccgtt
     2161 actccaagga gcacatgaag aagatgatga aggacttgga ggggttacac cgagctgagc
     2221 agagtctgca tgaccttcag gaaaggctgc acaaggccca ggaggagcac cgcacagtgg
     2281 aggtggagaa ggtccatctg gaaaagaagc tgcgcgatga gatcaacctt gctaagcagg
     2341 aagcccagcg gctgaaggag ctgcgggagg gtactgagaa tgagcggagc cgccaaaaat
     2401 atgctgagga ggagttggag caggttcggg aggccttgag gaaagcagag aaggagctag
     2461 aatctcacag ctcatggtat gctccagagg cccttcagaa gtggctgcag ctgacacatg
     2521 aggtggaggt gcaatattac aacatcaaga agcaaaatgc tgagaagcag ctgctggtgg
     2581 ccaaggaggg ggctgagaag ataaaaaaga agagaaacac actctttggc accttccacg
     2641 tggcccacag ctcttccctg gatgatgtag atcataaaat tctaacagct aagcaagcac
     2701 tgagcgaggt gacagcagca ttgcgggagc gcctgcaccg ctggcaacag atcgagatcc
     2761 tctgtggctt ccagattgtc aacaaccctg gcatccactc actggtggct gccctcaaca
     2821 tagaccccag ctggatgggc agtacacgcc ccaaccctgc tcacttcatc atgactgacg
     2881 acgtggatga catggatgag gagattgtgt ctcccttgtc catgcagtcc cctagcctgc
     2941 agagcagtgt tcggcagcgc ctgacggagc cacagcatgg cctgggatct cagagggatt
     3001 tgacccattc cgattcggag tcctccctcc acatgagtga ccgccagcgt gtggccccca
     3061 aacctcctca gatgagccgt gctgcagacg aggctctcaa tgccatgact tccaatggca
     3121 gccaccggct gatcgagggg gtccacccag ggtctctggt ggagaaactg cctgacagcc
     3181 ctgccctggc caagaaggca ttactggcgc tgaaccatgg gctggacaag gcccacagcc
     3241 tgatggagct gagcccctca gccccacctg gtggctctcc acatttggat tcttcccgtt
     3301 ctcacagccc cagctcccca gacccagaca caccatctcc agttggggac agccgagccc
     3361 tgcaagccag ccgaaacaca cgcaataacc acctggctgg caagaaggct gtggctgagg
     3421 aggataatgg ctctattggc gaggaaacag actccagccc aggccggaag aagtttcccc
     3481 tcaaaatctt taagaagcct cttaagaagt agggatccgc ggccgcatag ataactgatc
     3541 cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga gtactccctc
     3601 tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg
     3661 atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg gtcagaaaag
     3721 acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc atacacttga
     3781 gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
     3841 aggtcgagca tgcatctagg gcggccaatt ccgcccctct ccctcccccc cccctaacgt
     3901 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     3961 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     4021 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     4081 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     4141 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     4201 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     4261 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     4321 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     4381 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     4441 atgataagct tgccacaacc cgggaggcgc gccatgattc acaccaacct gaagaaaaag
     4501 ttcagctgct gcgtcctggt ctttcttctg tttgcagtca tctgtgtgtg gaaggaaaag
     4561 aagaaaggga gttactatga ttcctttaaa ttgcaaacca aggggaattc catggacgag
     4621 aagaccactg gttggcgagg tggacacgtt gttgaaggac tggctgggga acttgaacaa
     4681 cttcgtgcac gactggagca tcacccacaa ggtcaacgtg aaccacctgc aggtatggtg
     4741 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     4801 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4861 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     4921 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     4981 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     5041 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctaca ggacggcgag
     5101 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     5161 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     5221 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     5281 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     5341 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     5401 gagggccgcc actccaccgg cggcatggac gagctgtaca agggccggcc ttaaggcctc
     5461 gagggccaat taattaactc tagataactg atcataatca gccataccac atttgtagag
     5521 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5581 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5641 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5701 ctcatcaatg tatcttaacg cgtcgagtgc attctagttg tggtttgtcc aaactcatca
     5761 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5821 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5881 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5941 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6001 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6061 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6121 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6661 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6721 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6781 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6841 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6901 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6961 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7021 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7081 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7141 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7201 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7261 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7321 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7381 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7441 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7501 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7561 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7621 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7681 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7741 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7801 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7861 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7921 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
