LOCUS       sequence-121464-        8668 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 8668)
  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 8668)
  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-mCherry-Golgin84
FEATURES             Location/Qualifiers
     source          1..8668
                     /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             1459..1485
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2220..2449
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2432..2451
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2507..3081,3082..3084,3085..3093)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2674..2691)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2901..2920
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3121..3234
                     /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             3244..3948
                     /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(3386..3404)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3654..3673
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    6292..6413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6329..6348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6515..6531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6515..6531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6528..6550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6539..6555
                     /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(6563..6569,6570..6587,6588..6593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6608..6629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6746..6763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6917..7505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6997..7016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7676..8467,8468..8536))
                     /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     8299..8318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8537..8641)
                     /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 catgcaccgg aggagatcac gctcttgtag ggaggaccag
      961 aaacctgtca ccggtgaccc tagcaaagac tcaaaagctc aggtgtccgc tgccgaggct
     1021 ggcattactg gaacatggta caatcagctc gggagcacct ttattgtgac tgctggagcc
     1081 gatggagccc tcaccggaac atacgaatct gctgtgggaa acgccgaatc acggtacgtc
     1141 ctcactggcc gatacgatag tgcccctgcc accgacggat ctgggactgc cctgggatgg
     1201 actgtcgctt ggaaaaacaa ctaccggaat gctcattctg ccacaacatg gagtggacag
     1261 tacgtgggag gcgctgaggc tagaatcaat acacagtggc tgctcacatc tggcacaacc
     1321 gaggcaaatg cttggaaatc caccctggtg ggacatgaca cattcaccaa agtgaaaccc
     1381 tccgccgctt caatcgatgc cgccaaaaaa gccggagtca acaacggcaa tcctctggat
     1441 gccgtccagc aggtcgacta tccgtacgac gtaccagact acgcagtcgg accgatggac
     1501 gatcagaggg acctcattag caacaacgaa cagctgccta tgctgggacg gcgacctgga
     1561 gcccctgaat ccaaatgctc taggggagca ctgtacactg gcttctccat tctcgtgaca
     1621 ctgctgctgg ccgggcaggc tactactgct tacttcctgt accagcagca ggggcggctg
     1681 gacaaactca ctgtgacatc tcagaacctc cagctggaaa atctgaggat gaaactgccc
     1741 aaacccccta aacccgtgtc caaaatgagg atggccacac ctctgctcat gcaggcactg
     1801 ccaatgggag ccctgcccca ggggcccatg cagaatgcca ccaagtatgg caacatgaca
     1861 gaggaccatg tgatgcacct gctccagaat gctgaccccc tgaaggtgta cccgccactg
     1921 aaggggagct tcccggagaa cctgagacac cttaagaaca ccatggagac catagactgg
     1981 aaggtctttg agagctggat gcaccattgg ctcctgtttg aaatgagcag gcactccttg
     2041 gagcaaaagc ccactgacgc tccaccgaaa gagtcactgg aactggagga cccgtcttct
     2101 gggctgggtg tgaccaagca ggatctgggc ccagtcccca tgtgaggatc cgcggccgca
     2161 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg
     2221 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa
     2281 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat
     2341 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg
     2401 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     2461 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccctccc
     2521 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     2581 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     2641 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     2701 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     2761 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2821 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     2881 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     2941 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     3001 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     3061 tttgaaaaac acgatgataa gcttgccaca acccgggagg cgcgccaaat tgcgaattcc
     3121 atggacgaga agaccactgg ttggcgaggt ggacacgttg ttgaaggact ggctggggaa
     3181 cttgaacaac ttcgtgcacg actggagcat cacccacaag gtcaacgtga accacctgca
     3241 ggtgtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     3301 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     3361 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     3421 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     3481 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     3541 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctacaggac
     3601 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     3661 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     3721 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     3781 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     3841 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     3901 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg ccggccttct
     3961 tggtttgttg atcttgctgg aaaggcagaa gatcttttaa accgagttga tcaaggggct
     4021 gcaacagctc tcagtaggaa agacaatgcc agcaacatat atagcaaaaa tactgactat
     4081 actgaacttc accagcaaaa tacagatttg atatatcaga ctggacctaa atctacgtat
     4141 atttcatcag caggtgataa cattcgaaat caaaaagcca ccatcttagc tggcactgca
     4201 aatgtgaaag taggatctcg gacaccagta gaggcctctc atcctgttga aaatgcatct
     4261 gttcctaggc cttcatccca ttttgtgcga agaaaaaagt cagaacctga tgatgagctg
     4321 ctgtttgatt ttcttaatag ttcacagaag gagcctaccg ggagggtgga aatcagaaag
     4381 gaaaaaggca agacacctgt ctttcagagc tctcagacat caagtgtcag ttctgtgaac
     4441 cccagtgtaa ccaccatcaa aaccattgaa gaaaattctt ttgggagcca aacccacgaa
     4501 gctgccagta actcagattc tagccatgaa ggtcaagagg aatcttcaaa ggaaaatgtg
     4561 tcatcaaatg ctgcctgccc tgaccacacc ccaacaccta atgatgatgg caaatcacat
     4621 gaactgtcta accttcgact ggagaatcag ctgctgagga atgaagttca gtctttaaat
     4681 caagaaatgg cctcgttact ccaaagatcc aaagagactc aagaagaatt aaacaaagca
     4741 agagcaagag ttgaaaagtg gaatgctgac cattcaaaga gtgatcgaat gactcgagga
     4801 ctccgagccc aagtagatga cctgactgaa gctgtggctg caaaggattc ccagctggct
     4861 gtactgaaag tgagactcca ggaagctgac cagctactga gtactcgcac agaagcatta
     4921 gaagccttac agagtgaaaa atcacgaata atgcaggatc aaagtgaagg taacagcctg
     4981 cagaatcaag ctctgcagac ttttcaggag agactgcatg aagcggatgc cactctgaag
     5041 agagagcagg agagctataa acagatgcag agcgagtttg ctgcacgcct taataaagtg
     5101 gaaatggaac gtcagaattt agcagaagca attacactgg ccgaaagaaa atactcagat
     5161 gagaagaaga gggttgatga actgcagcag caagtcaagc tgtataagtt gaacttggag
     5221 tcctctaagc aggaattaat tgactacaag caaaaagcta ctagaatact gcaatctaag
     5281 gaaaaattga ttaacagctt gaaagaaggc tctggttttg aaggcctaga tagcagcact
     5341 gccagtagca tggagctgga agaacttcgg catgagaaag agatgcagag ggaggaaata
     5401 cagaagctga tgggccagat acatcagctc agatccgaat tacaggatat ggaggcacag
     5461 caagttaatg aagcagaatc agcaagagaa cagttacagg atctgcatga ccaaatagct
     5521 gggcagaaag catccaaaca agaactagag acagaactgg agcgactgaa gcaggagttc
     5581 cactatatag aagaagatct ttatcgaaca aagaacacat tgcaaagcag aattaaagat
     5641 cgagacgaag aaattcaaaa actcaggaat cagcttacca ataaaacttt aagcaatagc
     5701 agtcagtctg agttagaaaa tcgactccat cagctaacag agactctcat ccagaaacag
     5761 accatgctgg agagtctcag cacagaaaag aactccctgg tctttcaact ggagcgcctc
     5821 gaacagcaga tgaactccgc ctctggaagt agtagtaatg ggtcttcgat taatatgtct
     5881 ggaattgaca atggtgaagg cactcgtctg cgaaatgttc ctgttctttt taatgacaca
     5941 gaaactaatc tggcaggaat gtacggaaaa gttcgcaaag ctgctagttc aattgatcag
     6001 tttagtattc gcctgggaat ttttctccga agatacccca tagcgcgagt ttttgtaatt
     6061 atatatatgg ctttgcttca cctctgggtc atgattgttc tgttgactta cacaccagaa
     6121 atgcaccacg accaaccata tggcaaatga ggcctcgagg gccaattaat taactctaga
     6181 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     6241 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     6301 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6361 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc
     6421 gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6481 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6541 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6601 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6661 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6721 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6781 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6841 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6901 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6961 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7021 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7081 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     7141 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7201 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7261 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7321 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7381 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7441 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7501 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7561 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7621 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7681 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7741 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7801 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7861 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7921 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7981 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8041 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8101 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8161 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8221 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8281 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8341 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8401 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8461 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8521 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8581 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8641 gcacatttcc ccgaaaagtg ccacctga
//
