LOCUS       sequence-430451-        7790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luminal ER hook only (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7790)
  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 7790)
  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_neomycin
FEATURES             Location/Qualifiers
     source          1..7790
                     /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"
     intron          3591..3820
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3803..3822
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3893..4467,4468..4470,4471..4479)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4060..4077)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4287..4306
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4493..5287
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4493..5287
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4547..4566)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5157..5176
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5414..5535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5451..5470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5637..5653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5637..5653)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5650..5672)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5661..5677
                     /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(5685..5691,5692..5709,5710..5715))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5730..5751
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5868..5885)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6039..6627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6119..6138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6798..7589,7590..7658))
                     /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     7421..7440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7659..7763)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca 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 ggagtattta 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 aaccggtatg
      961 gatgtatgcg tccgtcttgc cctgtggctc ctctggggac tcctcctgca ccagggccag
     1021 agcctcagcc atagtcacag tgagaaggcg acaggaacca gctcgggcgc cggtttaaac
     1081 gaccctagca aagactcaaa agctcaggtg tccgctgccg aggctggcat tactggaaca
     1141 tggtacaatc agctcgggag cacctttatt gtgactgctg gagccgatgg agccctcacc
     1201 ggaacatacg aatctgctgt gggaaacgcc gaatcacggt acgtcctcac tggccgatac
     1261 gatagtgccc ctgccaccga cggatctggg actgccctgg gatggactgt cgcttggaaa
     1321 aacaactacc ggaatgctca ttctgccaca acatggagtg gacagtacgt gggaggcgct
     1381 gaggctagaa tcaatacaca gtggctgctc acatctggca caaccgaggc aaatgcttgg
     1441 aaatccaccc tggtgggaca tgacacattc accaaagtga aaccctccgc cgcttcaatc
     1501 gatgccgcca aaaaagccgg agtcaacaac ggcaatcctc tggatgccgt ccagcagctc
     1561 ggaccgggcg ccaactctga ggagtccact gcagcagagt tttgccgaat tgacaagccc
     1621 ctgtgtcaca gtgaggatga gaaactcagc ttcgaggcag tccgtaacat ccacaaactg
     1681 atggacgatg atgccaatgg tgatgtggat gtggaagaaa gtgatgagtt cctgagggaa
     1741 gacctcaatt accatgaccc aacagtgaaa cacagcacct tccatggtga ggataagctc
     1801 atcagcgtgg aggacctgtg gaaggcatgg aagtcatcag aagtatacaa ttggaccgtg
     1861 gatgaggtgg tacagtggct gatcacatat gtggagctgc ctcagtatga ggagaccttc
     1921 cggaagctgc agctcagtgg ccatgccatg ccaaggctgg ctgtcaccaa caccaccatg
     1981 acagggactg tgctgaagat gacagaccgg agtcatcggc agaagctgca gctgaaggct
     2041 ctggatacag tgctctttgg gcctcctctc ttgactcgcc ataatcacct caaggacttc
     2101 atgctggtgg tgtctatcgt tattggtgtg ggcggctgct ggtttgccta tatccagaac
     2161 cgttactcca aggagcacat gaagaagatg atgaaggact tggaggggtt acaccgagct
     2221 gagcagagtc tgcatgacct tcaggaaagg ctgcacaagg cccaggagga gcaccgcaca
     2281 gtggaggtgg agaaggtcca tctggaaaag aagctgcgcg atgagatcaa ccttgctaag
     2341 caggaagccc agcggctgaa ggagctgcgg gagggtactg agaatgagcg gagccgccaa
     2401 aaatatgctg aggaggagtt ggagcaggtt cgggaggcct tgaggaaagc agagaaggag
     2461 ctagaatctc acagctcatg gtatgctcca gaggcccttc agaagtggct gcagctgaca
     2521 catgaggtgg aggtgcaata ttacaacatc aagaagcaaa atgctgagaa gcagctgctg
     2581 gtggccaagg agggggctga gaagataaaa aagaagagaa acacactctt tggcaccttc
     2641 cacgtggccc acagctcttc cctggatgat gtagatcata aaattctaac agctaagcaa
     2701 gcactgagcg aggtgacagc agcattgcgg gagcgcctgc accgctggca acagatcgag
     2761 atcctctgtg gcttccagat tgtcaacaac cctggcatcc actcactggt ggctgccctc
     2821 aacatagacc ccagctggat gggcagtaca cgccccaacc ctgctcactt catcatgact
     2881 gacgacgtgg atgacatgga tgaggagatt gtgtctccct tgtccatgca gtcccctagc
     2941 ctgcagagca gtgttcggca gcgcctgacg gagccacagc atggcctggg atctcagagg
     3001 gatttgaccc attccgattc ggagtcctcc ctccacatga gtgaccgcca gcgtgtggcc
     3061 cccaaacctc ctcagatgag ccgtgctgca gacgaggctc tcaatgccat gacttccaat
     3121 ggcagccacc ggctgatcga gggggtccac ccagggtctc tggtggagaa actgcctgac
     3181 agccctgccc tggccaagaa ggcattactg gcgctgaacc atgggctgga caaggcccac
     3241 agcctgatgg agctgagccc ctcagcccca cctggtggct ctccacattt ggattcttcc
     3301 cgttctcaca gccccagctc cccagaccca gacacaccat ctccagttgg ggacagccga
     3361 gccctgcaag ccagccgaaa cacacgcaat aaccacctgg ctggcaagaa ggctgtggct
     3421 gaggaggata atggctctat tggcgaggaa acagactcca gcccaggccg gaagaagttt
     3481 cccctcaaaa tctttaagaa gcctcttaag aagtagggat ccgcggccgc atagataact
     3541 gatccagtgt gctggaatta attcgctgtc tgcgagggcc agctgttggg gtgagtactc
     3601 cctctcaaaa gcgggcatga cttctgcgct aagattgtca gtttccaaaa acgaggagga
     3661 tttgatattc acctggcccg cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga
     3721 aaagacaatc tttttgttgt caagcttgag gtgtggcagg cttgagatct ggccatacac
     3781 ttgagtgaca atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     3841 ctgcaggtcg agcatgcatc tagggcggcc aattccgccc ctctcccccc cccccctctc
     3901 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     3961 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     4021 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     4081 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     4141 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4201 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     4261 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     4321 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     4381 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     4441 ttttcctttg aaaaacacga tgataagctt gccacaaccc gggaggcgcg ccatgattga
     4501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4921 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5041 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5281 cttctgatta attaactcta gataactgat cataatcagc cataccacat ttgtagaggt
     5341 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5401 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5461 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5521 catcaatgta tcttaacgcg tcgagtgcat tctagttgtg gtttgtccaa actcatcaat
     5581 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5641 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5701 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5761 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5821 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5881 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5941 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6001 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6061 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6421 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6661 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6721 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6781 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6841 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6901 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6961 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7021 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7081 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7141 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7201 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7261 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7321 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7381 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7441 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7501 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7561 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7621 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7681 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7741 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
//
