LOCUS       sequence-120697-        7985 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 7985)
  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 7985)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..7985
                     /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             join(4735..4737,4738..4740,4741..5451)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4735..4737,4738..4740,4741..5451)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4780..4801)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5041..5060)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5388..5409
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5605..5726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5642..5661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5828..5844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5828..5844)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5841..5863)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5852..5868
                     /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(5876..5882,5883..5900,5901..5906))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5921..5942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6059..6076)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6230..6818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6310..6329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6989..7780,7781..7849))
                     /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     7612..7631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7850..7954)
                     /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 agcaaaggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     4801 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     4861 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4921 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     4981 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     5041 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     5101 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     5161 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     5221 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     5281 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     5341 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     5401 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggccggcct
     5461 taaggcctcg agggccaatt aattaactct agataactga tcataatcag ccataccaca
     5521 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5581 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5641 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5701 ttgtccaaac tcatcaatgt atcttaacgc gtcgagtgca ttctagttgt ggtttgtcca
     5761 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5821 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5881 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5941 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6001 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6061 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6421 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6661 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7981 acgtc
//
