LOCUS       sequence-385985-        8692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synchronize trafficking of Golgin-84 from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8692)
  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 8692)
  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-Ii_SBP-EGFP-Golgin84
FEATURES             Location/Qualifiers
     source          1..8692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1221..1247
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          1982..2211
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2194..2213
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2284..2858,2859..2861,2862..2870)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2451..2468)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2678..2697
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2898..3011
                     /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(3021..3023,3024..3734)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3021..3023,3024..3734)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3063..3084)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3324..3343)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3671..3692
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6078..6199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6115..6134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6301..6317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6301..6317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6314..6336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6325..6341
                     /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(6349..6355,6356..6373,6374..6379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6394..6415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6532..6549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6703..7291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6783..6802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7462..8253,8254..8322))
                     /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     8085..8104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8323..8427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8502..8521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcgat atcatgcacc ggaggagatc acgctcttgt agggaggacc
      721 agaaacctgt caccggtgac cctagcaaag actcaaaagc tcaggtgtcc gctgccgagg
      781 ctggcattac tggaacatgg tacaatcagc tcgggagcac ctttattgtg actgctggag
      841 ccgatggagc cctcaccgga acatacgaat ctgctgtggg aaacgccgaa tcacggtacg
      901 tcctcactgg ccgatacgat agtgcccctg ccaccgacgg atctgggact gccctgggat
      961 ggactgtcgc ttggaaaaac aactaccgga atgctcattc tgccacaaca tggagtggac
     1021 agtacgtggg aggcgctgag gctagaatca atacacagtg gctgctcaca tctggcacaa
     1081 ccgaggcaaa tgcttggaaa tccaccctgg tgggacatga cacattcacc aaagtgaaac
     1141 cctccgccgc ttcaatcgat gccgccaaaa aagccggagt caacaacggc aatcctctgg
     1201 atgccgtcca gcaggtcgac tatccgtacg acgtaccaga ctacgcagtc ggaccgatgg
     1261 acgatcagag ggacctcatt agcaacaacg aacagctgcc tatgctggga cggcgacctg
     1321 gagcccctga atccaaatgc tctaggggag cactgtacac tggcttctcc attctcgtga
     1381 cactgctgct ggccgggcag gctactactg cttacttcct gtaccagcag caggggcggc
     1441 tggacaaact cactgtgaca tctcagaacc tccagctgga aaatctgagg atgaaactgc
     1501 ccaaaccccc taaacccgtg tccaaaatga ggatggccac acctctgctc atgcaggcac
     1561 tgccaatggg agccctgccc caggggccca tgcagaatgc caccaagtat ggcaacatga
     1621 cagaggacca tgtgatgcac ctgctccaga atgctgaccc cctgaaggtg tacccgccac
     1681 tgaaggggag cttcccggag aacctgagac accttaagaa caccatggag accatagact
     1741 ggaaggtctt tgagagctgg atgcaccatt ggctcctgtt tgaaatgagc aggcactcct
     1801 tggagcaaaa gcccactgac gctccaccga aagagtcact ggaactggag gacccgtctt
     1861 ctgggctggg tgtgaccaag caggatctgg gcccagtccc catgtgagga tccgcggccg
     1921 catagataac tgatccagtg tgctggaatt aattcgctgt ctgcgagggc cagctgttgg
     1981 ggtgagtact ccctctcaaa agcgggcatg acttctgcgc taagattgtc agtttccaaa
     2041 aacgaggagg atttgatatt cacctggccc gcggtgatgc ctttgagggt ggccgcgtcc
     2101 atctggtcag aaaagacaat ctttttgttg tcaagcttga ggtgtggcag gcttgagatc
     2161 tggccataca cttgagtgac aatgacatcc actttgcctt tctctccaca ggtgtccact
     2221 cccaggtcca actgcaggtc gagcatgcat ctagggcggc caattccgcc cctctccccc
     2281 ccccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     2341 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     2401 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     2461 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     2521 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     2581 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     2641 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     2701 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     2761 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     2821 cggggacgtg gttttccttt gaaaaacacg atgataagct tgccacaacc cgggaggcgc
     2881 gccaaattgc gaattccatg gacgagaaga ccactggttg gcgaggtgga cacgttgttg
     2941 aaggactggc tggggaactt gaacaacttc gtgcacgact ggagcatcac ccacaaggtc
     3001 aacgtgaacc acctgcaggt gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3061 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3121 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3181 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3241 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3301 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3361 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3421 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3481 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3541 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3601 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3661 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3721 acgagctgta caagggccgg ccttcttggt ttgttgatct tgctggaaag gcagaagatc
     3781 ttttaaaccg agttgatcaa ggggctgcaa cagctctcag taggaaagac aatgccagca
     3841 acatatatag caaaaatact gactatactg aacttcacca gcaaaataca gatttgatat
     3901 atcagactgg acctaaatct acgtatattt catcagcagg tgataacatt cgaaatcaaa
     3961 aagccaccat cttagctggc actgcaaatg tgaaagtagg atctcggaca ccagtagagg
     4021 cctctcatcc tgttgaaaat gcatctgttc ctaggccttc atcccatttt gtgcgaagaa
     4081 aaaagtcaga acctgatgat gagctgctgt ttgattttct taatagttca cagaaggagc
     4141 ctaccgggag ggtggaaatc agaaaggaaa aaggcaagac acctgtcttt cagagctctc
     4201 agacatcaag tgtcagttct gtgaacccca gtgtaaccac catcaaaacc attgaagaaa
     4261 attcttttgg gagccaaacc cacgaagctg ccagtaactc agattctagc catgaaggtc
     4321 aagaggaatc ttcaaaggaa aatgtgtcat caaatgctgc ctgccctgac cacaccccaa
     4381 cacctaatga tgatggcaaa tcacatgaac tgtctaacct tcgactggag aatcagctgc
     4441 tgaggaatga agttcagtct ttaaatcaag aaatggcctc gttactccaa agatccaaag
     4501 agactcaaga agaattaaac aaagcaagag caagagttga aaagtggaat gctgaccatt
     4561 caaagagtga tcgaatgact cgaggactcc gagcccaagt agatgacctg actgaagctg
     4621 tggctgcaaa ggattcccag ctggctgtac tgaaagtgag actccaggaa gctgaccagc
     4681 tactgagtac tcgcacagaa gcattagagg ccttacagag tgaaaaatca cgaataatgc
     4741 aggatcaaag tgaaggtaac agcctgcaga atcaagctct gcagactctt caggagagac
     4801 tgcatgaagc ggatgccact ctgaagagag agcaggagag ctataaacag atgcagagcg
     4861 agtttgctgc acgccttaat aaagtggaaa tggaacgtca gaatttagca gaagcaatta
     4921 cactggccga aagaaaatac tcagatgaga agaagagggt tgatgaactg cagcagcaag
     4981 tcaagctgta taagttgaac ttggagtcct ctaagcagga attaattgac tacaagcaaa
     5041 aagctactag aatactgcaa tctaaggaaa aattgattaa cagcttgaaa gaaggctctg
     5101 gttttgaagg cctagatagc agcactgcca gtagcatgga gctggaagaa cttcggcatg
     5161 agaaagagat gcagagggag gaaatacaga agctgatggg ccagatacat cagctcagat
     5221 ccgaattaca ggatatggag gcacagcaag ttaatgaagc agaatcagca agagaacagt
     5281 tacaggatct gcatgaccaa atagctgggc agaaagcatc caaacaagaa ctagagacag
     5341 aactggagcg actgaagcag gagttccact atatagaaga agatctttat cgaacaaaga
     5401 acacattgca aagcagaatt aaagatcgag acgaagaaat tcaaaaactc aggaatcagc
     5461 ttaccaataa aactttaagc aatagcagtc agtctgagtt agaaaatcga ctccatcagc
     5521 taacagagac tctcatccag aaacagacca tgctggagag tctcagcaca gaaaagaact
     5581 ccctggtctt tcaactggag cgcctcgaac agcagatgaa ctccgcctct ggaagtagta
     5641 gtaatgggtc ttcgattaat atgtctggaa ttgacaatgg tgaaggcact cgtctgcgaa
     5701 atgttcctgt tctttttaat gacacagaaa ctaatctggc aggaatgtac ggaaaagttc
     5761 gcaaagctgc tagttcaatt gatcagttta gtattcgcct gggaattttt ctccgaagat
     5821 accccatagc gcgagttttt gtaattatat atatggcttt gcttcacctc tgggtcatga
     5881 ttgttctgtt gacttacaca ccagaaatgc accacgacca accatatggc aaatgaggcc
     5941 tcgagggcca attaattaac tctagataac tgatcataat cagccatacc acatttgtag
     6001 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     6061 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6181 aactcatcaa tgtatcttaa cgcgtcgagt gcattctagt tgtggtttgt ccaaactcat
     6241 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8461 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     8521 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8581 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8641 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
