LOCUS       sequence-110517-        6415 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 6415)
  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 6415)
  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: Ii-Str_puromycin
FEATURES             Location/Qualifiers
     source          1..6415
                     /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"
     CDS             1749..1775
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2294..2523
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2506..2525
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2596..3170,3171..3173,3174..3182)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2763..2780)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2990..3009
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3202..3801
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3202..3221)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3698..3718
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4035..4156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4072..4091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4258..4274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4258..4274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4271..4293)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4282..4298
                     /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(4306..4312,4313..4330,4331..4336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4351..4372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4489..4506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4660..5248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4740..4759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5419..6210,6211..6279))
                     /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     6042..6061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6280..6384)
                     /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 ggtcgtacgt
      961 ctccggattc gaattaattc ggatccgtga tgcacaggag gagagccagg gcctgtcggg
     1021 aagatcaaaa gccagtcatc gatgatcagc gcgaccttat ctccaacaat gagcaactgc
     1081 ccatgctggg ccggcgccct ggggccccgg agagcaagtg cagccgcgga gccctgtaca
     1141 caggcttttc catcctggtg actctgctcc tcgctggcca ggccaccacc gcctacttcc
     1201 tgtaccagca gcagggccgg ctggacaaac tgacagtcac ctcccagaac ctgcagctcg
     1261 agatggaccc tagcaaagac tcaaaagctc aggtgtccgc tgccgaggct ggcattactg
     1321 gaacatggta caatcagctc gggagcacct ttattgtgac tgctggagcc gatggagccc
     1381 tcaccggaac atacgaatct gctgtgggaa acgccgaatc acggtacgtc ctcactggcc
     1441 gatacgatag tgcccctgcc accgacggat ctgggactgc cctgggatgg actgtcgctt
     1501 ggaaaaacaa ctaccggaat gctcattctg ccacaacatg gagtggacag tacgtgggag
     1561 gcgctgaggc tagaatcaat acacagtggc tgctcacatc tggcacaacc gaggcaaatg
     1621 cttggaaatc caccctggtg ggacatgaca cattcaccaa agtgaaaccc tccgccgctt
     1681 caatcgatgc cgccaaaaaa gccggagtca acaacggcaa tcctctggat gccgtccagc
     1741 aggtcgacta tccgtacgac gtaccagact acgcactcga cctgcagctg gagaacctgc
     1801 gcatgaagct tcccaagcct cccaagcctg tgagcaagat gcgcatggcc accccgctgc
     1861 tgatgcaggc gctgcccatg ggagccctgc cccaggggcc catgcagaat gccaccaagt
     1921 atggcaacat gacagaggac catgtgatgc acctgctcca gaatgctgac cccctgaagg
     1981 tgtacccgcc actgaagggg agcttcccgg agaacctgag acaccttaag aacaccatgg
     2041 agaccataga ctggaaggtc tttgagagct ggatgcacca ttggctcctg tttgaaatga
     2101 gcaggcactc cttggagcaa aagcccactg acgctccacc gaaagagtca ctggaactgg
     2161 aggacccgtc ttctgggctg ggtgtgacca agcaggatct gggcccagtc cccatgtgag
     2221 gatccgcggc cgcatagata actgatccag tgtgctggaa ttaattcgct gtctgcgagg
     2281 gccagctgtt ggggtgagta ctccctctca aaagcgggca tgacttctgc gctaagattg
     2341 tcagtttcca aaaacgagga ggatttgata ttcacctggc ccgcggtgat gcctttgagg
     2401 gtggccgcgt ccatctggtc agaaaagaca atctttttgt tgtcaagctt gaggtgtggc
     2461 aggcttgaga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
     2521 caggtgtcca ctcccaggtc caactgcagg tcgagcatgc atctagggcg gccaattccg
     2581 cccctctccc ccccccccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     2641 ggaataaggc cggtgtgcgt ttgtctatat gtgattttcc accatattgc cgtcttttgg
     2701 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     2761 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     2821 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     2881 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     2941 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     3001 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     3061 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     3121 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa
     3181 cccacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3241 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3301 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3361 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3421 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3481 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     3541 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     3601 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     3661 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     3721 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     3781 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     3841 gcacgacccc atggctccga ccgaagccac ccggggcggc cccgccgacc ccgcacccgc
     3901 ccccgaggcc caccgactct agataactga tcataatcag ccataccaca tttgtagagg
     3961 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4021 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4081 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4141 tcatcaatgt atcttaacgc gtcgagtgca ttctagttgt ggtttgtcca aactcatcaa
     4201 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4261 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     4861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5101 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5701 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6001 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6241 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6301 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
