LOCUS       sequence-120701-        8147 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 8147)
  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 8147)
  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-EGFP-Ecadherin
FEATURES             Location/Qualifiers
     source          1..8147
                     /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             join(2713..2715,2716..2718,2719..3429)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2713..2715,2716..2718,2719..3429)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2758..2779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3019..3038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3366..3387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5767..5888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5804..5823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5990..6006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5990..6006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6003..6025)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6014..6030
                     /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(6038..6044,6045..6062,6063..6068))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6083..6104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6221..6238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6392..6980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6472..6491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7151..7942,7943..8011))
                     /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     7774..7793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8012..8116)
                     /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 gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2761 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2821 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2881 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2941 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3001 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3061 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3121 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3181 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3241 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3301 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3361 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3421 ctgtacaagg gccggccaga ctgggtcatc cctcccatca gctgccccga aaatgaaaag
     3481 ggtgaattcc caaagaacct ggttcagatc aaatccaaca gggacaaaga aacaaaggtt
     3541 ttctacagca tcaccggcca aggagctgac aaaccccccg ttggcgtttt catcattgag
     3601 agggagacag gctggctgaa agtgacacag cctctggata gagaagccat tgccaagtac
     3661 atcctctatt ctcatgccgt gtcatcaaat ggggaagcgg tggaggatcc catggagata
     3721 gtgatcacag tgacagatca gaatgacaac aggccagagt ttacccagga ggtgtttgag
     3781 ggatccgttg cagaaggcgc tgttccagga acctccgtga tgaaggtctc agccaccgat
     3841 gcagacgatg acgtcaacac ctacaacgct gccatcgcct acaccatcgt cagccaggat
     3901 cctgagctgc ctcacaaaaa catgttcact gtcaataggg acaccggggt catcagtgtg
     3961 ctcacctctg ggctggaccg agagagttac cctacataca ctctggtggt tcaggctgct
     4021 gaccttcaag gcgaaggctt gagcacaaca gccaaggctg tgatcactgt caaggatatt
     4081 aatgacaacg ctcctgtctt caacccgagc acgtatcagg gtcaagtgcc tgagaatgag
     4141 gtcaatgccc ggatcgccac actcaaagtg accgatgatg atgcccccaa cactccggcg
     4201 tggaaagctg tgtacaccgt agtcaacgat cctgaccagc agttcgttgt tgtcacagac
     4261 cccacgacca atgatggcat tttgaaaaca gccaagggct tggattttga ggccaagcag
     4321 caatacatcc ttcatgtgag agtggagaac gaggaaccct ttgaggggtc tcttgtccct
     4381 tccacagcca ctgtcactgt ggacgtggta gacgtgaatg aagcccccat ctttatgcct
     4441 gcggagagga gagtcgaagt gcccgaagac tttggtgtgg gtcaggaaat cacatcttat
     4501 accgctcgag agccggacac gttcatggat cagaagatca cgtatcggat ttggagggac
     4561 actgccaact ggctggagat taacccagag actggtgcca ttttcacgcg cgctgagatg
     4621 gacagagaag acgctgagca tgtgaagaac agcacatatg tagctctcat catcgccaca
     4681 gatgatggtt cacccattgc cactggcacg ggcactcttc tcctggtcct gttagacgtc
     4741 aatgacaacg ctcccatccc agaacctcga aacatgcagt tctgccagag gaacccacag
     4801 cctcatatca tcaccatctt ggatccagac cttcccccca acacgtcccc ctttactgct
     4861 gagctaaccc atggggccag cgtcaactgg accattgagt ataatgacgc agctcaagaa
     4921 tctctcattt tgcaaccaag aaaggactta gagattggcg aatacaaaat ccatctcaag
     4981 ctcgcggata accagaacaa agaccaggtg accacgttgg acgtccatgt gtgtgactgt
     5041 gaagggacgg tcaacaactg catgaaggcg ggaatcgtgg cagcaggatt gcaagttcct
     5101 gccatcctcg gaatccttgg agggatcctc gccctgctga ttctgatcct gctgctccta
     5161 ctgtttctac ggaggagaac ggtggtcaaa gagcccctgc tgccaccaga tgatgatacc
     5221 cgggacaatg tgtattacta tgatgaagaa ggaggtggag aagaagacca ggactttgat
     5281 ttgagccagc tgcacagggg cctggatgcc cgaccggaag tgactcgaaa tgatgtggct
     5341 cccaccctca tgagcgtgcc ccagtatcgt ccccgtcctg ccaatcctga tgaaattgga
     5401 aacttcatcg atgaaaacct gaaggcagcc gacagcgacc ccacggcacc cccttacgac
     5461 tctctgttgg tgttcgatta cgagggcagt ggttctgaag ccgctagcct gagctcactg
     5521 aactcctctg agtcggatca ggaccaggac tacgattatc tgaacgagtg gggcaaccga
     5581 ttcaagaagc tggcggacat gtacggcggt ggcgaagacg actagggcct cgagggccaa
     5641 ttaattaact ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     5701 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5761 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5821 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5881 gtatcttaac gcgtcgagtg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5941 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6001 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6061 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6121 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6181 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6241 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6301 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6361 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6421 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6481 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6541 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     6601 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6661 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6721 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6781 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6841 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6901 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6961 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7021 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7081 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7141 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7201 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7261 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7321 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7381 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7441 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7501 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7561 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7621 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7681 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7741 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7801 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7861 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7921 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7981 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8041 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8101 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
