LOCUS Exported 7985 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synchronize trafficking of ST from the ER (RUSH system). ACCESSION . VERSION . KEYWORDS Str-STIM1-NN_ST-SBP-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7985) 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 7985) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7985 /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 3587..3816 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 3799..3818 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 3874..4447 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4041..4058) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4268..4287 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4612..4725 /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="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP" CDS 4735..5451 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4780..4801) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5041..5060) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5388..5409 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 5605..5726 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5642..5661) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(5828..5844) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5828..5844) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5841..5863) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5852..5868 /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(5876..5906) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5921..5942 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6059..6076) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6230..6818) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6310..6329) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6989..7849) /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 7612..7631 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7850..7954) /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 cgggcgccaa ctctgaggag tccactgcag cagagttttg ccgaattgac aagcccctgt 1621 gtcacagtga ggatgagaaa ctcagcttcg aggcagtccg taacatccac aaactgatgg 1681 acgatgatgc caatggtgat gtggatgtgg aagaaagtga tgagttcctg agggaagacc 1741 tcaattacca tgacccaaca gtgaaacaca gcaccttcca tggtgaggat aagctcatca 1801 gcgtggagga cctgtggaag gcatggaagt catcagaagt atacaattgg accgtggatg 1861 aggtggtaca gtggctgatc acatatgtgg agctgcctca gtatgaggag accttccgga 1921 agctgcagct cagtggccat gccatgccaa ggctggctgt caccaacacc accatgacag 1981 ggactgtgct gaagatgaca gaccggagtc atcggcagaa gctgcagctg aaggctctgg 2041 atacagtgct ctttgggcct cctctcttga ctcgccataa tcacctcaag gacttcatgc 2101 tggtggtgtc tatcgttatt ggtgtgggcg gctgctggtt tgcctatatc cagaaccgtt 2161 actccaagga gcacatgaag aagatgatga aggacttgga ggggttacac cgagctgagc 2221 agagtctgca tgaccttcag gaaaggctgc acaaggccca ggaggagcac cgcacagtgg 2281 aggtggagaa ggtccatctg gaaaagaagc tgcgcgatga gatcaacctt gctaagcagg 2341 aagcccagcg gctgaaggag ctgcgggagg gtactgagaa tgagcggagc cgccaaaaat 2401 atgctgagga ggagttggag caggttcggg aggccttgag gaaagcagag aaggagctag 2461 aatctcacag ctcatggtat gctccagagg cccttcagaa gtggctgcag ctgacacatg 2521 aggtggaggt gcaatattac aacatcaaga agcaaaatgc tgagaagcag ctgctggtgg 2581 ccaaggaggg ggctgagaag ataaaaaaga agagaaacac actctttggc accttccacg 2641 tggcccacag ctcttccctg gatgatgtag atcataaaat tctaacagct aagcaagcac 2701 tgagcgaggt gacagcagca ttgcgggagc gcctgcaccg ctggcaacag atcgagatcc 2761 tctgtggctt ccagattgtc aacaaccctg gcatccactc actggtggct gccctcaaca 2821 tagaccccag ctggatgggc agtacacgcc ccaaccctgc tcacttcatc atgactgacg 2881 acgtggatga catggatgag gagattgtgt ctcccttgtc catgcagtcc cctagcctgc 2941 agagcagtgt tcggcagcgc ctgacggagc cacagcatgg cctgggatct cagagggatt 3001 tgacccattc cgattcggag tcctccctcc acatgagtga ccgccagcgt gtggccccca 3061 aacctcctca gatgagccgt gctgcagacg aggctctcaa tgccatgact tccaatggca 3121 gccaccggct gatcgagggg gtccacccag ggtctctggt ggagaaactg cctgacagcc 3181 ctgccctggc caagaaggca ttactggcgc tgaaccatgg gctggacaag gcccacagcc 3241 tgatggagct gagcccctca gccccacctg gtggctctcc acatttggat tcttcccgtt 3301 ctcacagccc cagctcccca gacccagaca caccatctcc agttggggac agccgagccc 3361 tgcaagccag ccgaaacaca cgcaataacc acctggctgg caagaaggct gtggctgagg 3421 aggataatgg ctctattggc gaggaaacag actccagccc aggccggaag aagtttcccc 3481 tcaaaatctt taagaagcct cttaagaagt agggatccgc ggccgcatag ataactgatc 3541 cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga gtactccctc 3601 tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg 3661 atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg gtcagaaaag 3721 acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc atacacttga 3781 gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc 3841 aggtcgagca tgcatctagg gcggccaatt ccgcccctct ccctcccccc cccctaacgt 3901 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac 3961 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag 4021 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa 4081 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag 4141 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga 4201 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag 4261 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc 4321 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag 4381 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg 4441 atgataagct tgccacaacc cgggaggcgc gccatgattc acaccaacct gaagaaaaag 4501 ttcagctgct gcgtcctggt ctttcttctg tttgcagtca tctgtgtgtg gaaggaaaag 4561 aagaaaggga gttactatga ttcctttaaa ttgcaaacca aggggaattc catggacgag 4621 aagaccactg gttggcgagg tggacacgtt gttgaaggac tggctgggga acttgaacaa 4681 cttcgtgcac gactggagca tcacccacaa ggtcaacgtg aaccacctgc aggtatggtg 4741 agcaaaggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 4801 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 4861 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 4921 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 4981 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 5041 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 5101 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 5161 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 5221 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 5281 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 5341 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 5401 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggccggcct 5461 taaggcctcg agggccaatt aattaactct agataactga tcataatcag ccataccaca 5521 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat 5581 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa 5641 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 5701 ttgtccaaac tcatcaatgt atcttaacgc gtcgagtgca ttctagttgt ggtttgtcca 5761 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5821 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5881 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5941 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6001 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 6061 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 6121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 6181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 6361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 6421 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 6481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 6541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6661 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 6781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 7021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 7081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 7141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 7201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 7261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 7321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 7381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 7441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 7501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 7561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 7621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7981 acgtc //