LOCUS       sequence-120702-        8135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synchronize trafficking of Ecadherin from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8135)
  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 8135)
  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-KDEL_SBP-mCherry-Ecadherin
FEATURES             Location/Qualifiers
     source          1..8135
                     /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          1646..1875
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1858..1877
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1933..2507,2508..2510,2511..2519)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2100..2117)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2327..2346
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     2533..2592
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             2593..2703
                     /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="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             2713..3417
                     /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(2855..2873)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3123..3142
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5755..5876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5792..5811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5978..5994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5978..5994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5991..6013)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6002..6018
                     /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(6026..6032,6033..6050,6051..6056))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6071..6092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6209..6226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6380..6968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6460..6479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7139..7930,7931..7999))
                     /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     7762..7781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8000..8104)
                     /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 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
     1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     1921 cggccaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     1981 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2041 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2101 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2161 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2221 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2281 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2341 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2401 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     2461 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     2521 gggaggcgcg ccatgtacag gatgcaactc ctgtcttgca ttgcactaag tcttgcactt
     2581 gtcacgaatt ccgacgagaa gaccactggt tggcgaggtg gacacgttgt tgaaggactg
     2641 gctggggaac ttgaacaact tcgtgcacga ctggagcatc acccacaagg tcaacgtgaa
     2701 ccacctgcag gtgtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     2761 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     2821 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     2881 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     2941 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     3001 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     3061 ctacaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     3121 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     3181 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     3241 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     3301 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     3361 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagggc
     3421 cggccagact gggtcatccc tcccatcagc tgccccgaaa atgaaaaggg tgaattccca
     3481 aagaacctgg ttcagatcaa atccaacagg gacaaagaaa caaaggtttt ctacagcatc
     3541 accggccaag gagctgacaa accccccgtt ggcgttttca tcattgagag ggagacaggc
     3601 tggctgaaag tgacacagcc tctggataga gaagccattg ccaagtacat cctctattct
     3661 catgccgtgt catcaaatgg ggaagcggtg gaggatccca tggagatagt gatcacagtg
     3721 acagatcaga atgacaacag gccagagttt acccaggagg tgtttgaggg atccgttgca
     3781 gaaggcgctg ttccaggaac ctccgtgatg aaggtctcag ccaccgatgc agacgatgac
     3841 gtcaacacct acaacgctgc catcgcctac accatcgtca gccaggatcc tgagctgcct
     3901 cacaaaaaca tgttcactgt caatagggac accggggtca tcagtgtgct cacctctggg
     3961 ctggaccgag agagttaccc tacatacact ctggtggttc aggctgctga ccttcaaggc
     4021 gaaggcttga gcacaacagc caaggctgtg atcactgtca aggatattaa tgacaacgct
     4081 cctgtcttca acccgagcac gtatcagggt caagtgcctg agaatgaggt caatgcccgg
     4141 atcgccacac tcaaagtgac cgatgatgat gcccccaaca ctccggcgtg gaaagctgtg
     4201 tacaccgtag tcaacgatcc tgaccagcag ttcgttgttg tcacagaccc cacgaccaat
     4261 gatggcattt tgaaaacagc caagggcttg gattttgagg ccaagcagca atacatcctt
     4321 catgtgagag tggagaacga ggaacccttt gaggggtctc ttgtcccttc cacagccact
     4381 gtcactgtgg acgtggtaga cgtgaatgaa gcccccatct ttatgcctgc ggagaggaga
     4441 gtcgaagtgc ccgaagactt tggtgtgggt caggaaatca catcttatac cgctcgagag
     4501 ccggacacgt tcatggatca gaagatcacg tatcggattt ggagggacac tgccaactgg
     4561 ctggagatta acccagagac tggtgccatt ttcacgcgcg ctgagatgga cagagaagac
     4621 gctgagcatg tgaagaacag cacatatgta gctctcatca tcgccacaga tgatggttca
     4681 cccattgcca ctggcacggg cactcttctc ctggtcctgt tagacgtcaa tgacaacgct
     4741 cccatcccag aacctcgaaa catgcagttc tgccagagga acccacagcc tcatatcatc
     4801 accatcttgg atccagacct tccccccaac acgtccccct ttactgctga gctaacccat
     4861 ggggccagcg tcaactggac cattgagtat aatgacgcag ctcaagaatc tctcattttg
     4921 caaccaagaa aggacttaga gattggcgaa tacaaaatcc atctcaagct cgcggataac
     4981 cagaacaaag accaggtgac cacgttggac gtccatgtgt gtgactgtga agggacggtc
     5041 aacaactgca tgaaggcggg aatcgtggca gcaggattgc aagttcctgc catcctcgga
     5101 atccttggag ggatcctcgc cctgctgatt ctgatcctgc tgctcctact gtttctacgg
     5161 aggagaacgg tggtcaaaga gcccctgctg ccaccagatg atgatacccg ggacaatgtg
     5221 tattactatg atgaagaagg aggtggagaa gaagaccagg actttgattt gagccagctg
     5281 cacaggggcc tggatgcccg accggaagtg actcgaaatg atgtggctcc caccctcatg
     5341 agcgtgcccc agtatcgtcc ccgtcctgcc aatcctgatg aaattggaaa cttcatcgat
     5401 gaaaacctga aggcagccga cagcgacccc acggcacccc cttacgactc tctgttggtg
     5461 ttcgattacg agggcagtgg ttctgaagcc gctagcctga gctcactgaa ctcctctgag
     5521 tcggatcagg accaggacta cgattatctg aacgagtggg gcaaccgatt caagaagctg
     5581 gcggacatgt acggcggtgg cgaagacgac tagggcctcg agggccaatt aattaactct
     5641 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     5701 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     5761 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5821 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     5881 gtcgagtgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5941 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6001 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6061 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6121 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6181 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6241 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6301 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6361 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6421 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6481 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6541 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     6601 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6661 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6721 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6781 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6841 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6901 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6961 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7021 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7081 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7141 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7201 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7261 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7321 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7381 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7441 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7501 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7561 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7621 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7681 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7741 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7801 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7861 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7921 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7981 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8041 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8101 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
