LOCUS Exported 8147 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synchronize trafficking of Ecadherin from the ER (RUSH system). ACCESSION . VERSION . KEYWORDS Str-KDEL_SBP-EGFP-Ecadherin 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8147 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1933..2506 /label=IRES /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 /product="human interleukin-2 signal sequence" /label=IL-2 signal sequence 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..3429 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2713..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(6038..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(7151..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 //