LOCUS Exported 7109 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synchronize trafficking of GalT from the ER (RUSH system). ACCESSION . VERSION . KEYWORDS Str-KDEL_flGalT-SBP-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7109) 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 7109) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7109 /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" CDS 3736..3849 /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 3859..4575 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3859..4575 /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(3904..3925) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4165..4184) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4512..4533 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4729..4850 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4766..4785) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4952..4968) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4952..4968) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4965..4987) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4976..4992 /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(5000..5030) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5045..5066 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5183..5200) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5354..5942) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5434..5453) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6113..6973) /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 6736..6755 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6974..7078) /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 ccatgaggct tcgggagccg ctcctgagcg gcagcgccgc gatgccaggc 2581 gcgtccctac agcgggcctg ccgcctgctc gtggccgtct gcgctctgca ccttggcgtc 2641 accctcgttt actacctggc tggccgcgac ctgagccgcc tgccccaact ggtcggagtc 2701 tccacaccgc tgcagggcgg ctcgaacagt gccgccgcca tcgggcagtc ctccggggag 2761 ctccggaccg gaggggcccg gccgccgcct cctctaggcg cctcctccca gccgcgcccg 2821 ggtggcgact ccagcccagt cgtggattct ggccctggcc ccgctagcaa cttgacctcg 2881 gtcccagtgc cccacaccac cgcactgtcg ctgcccgcct gccctgagga gtccccgctg 2941 cttgtgggcc ccatgctgat tgagtttaac atgcctgtgg acctggagct cgtggcaaag 3001 cagaacccaa atgtgaagat gggcggccgc tatgccccca gggactgcgt ctctcctcac 3061 aaggtggcca tcatcattcc attccgcaac cggcaggagc acctcaagta ctggctatat 3121 tatttgcacc cagtcctgca gcgccagcag ctggactatg gcatctatgt tatcaaccag 3181 gcgggagaca ctatattcaa tcgtgctaag ctcctcaatg ttggctttca agaagccttg 3241 aaggactatg actacacctg ctttgtgttt agtgacgtgg acctcattcc aatgaatgac 3301 cataatgcgt acaggtgttt ttcacagcca cggcacattt ccgttgcaat ggataagttt 3361 ggattcagcc taccttatgt tcagtatttt ggaggtgtct ctgctctaag taaacaacag 3421 tttctaacca tcaatggatt tcctaataat tattggggct ggggaggaga agatgatgac 3481 atttttaaca gattagtttt tagaggcatg tctatatctc gcccaaatgc tgtggtcggg 3541 aggtgtcgca tgatccgcca ctcaagagac aagaaaaatg aacccaatcc tcagaggttt 3601 gaccgaattg cacacacaaa ggagacaatg ctctctgatg gtttgaactc actcacctac 3661 caggtgctgg atgtacagag atacccattg tatacccaaa tcacagtgga catcgggaca 3721 ccgagcttga attccatgga cgagaagacc actggttggc gaggtggaca cgttgttgaa 3781 ggactggctg gggaacttga acaacttcgt gcacgactgg agcatcaccc acaaggtcaa 3841 cgtgaaccac ctgcaggtat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 3901 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 3961 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 4021 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc 4081 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 4141 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 4201 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 4261 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg 4321 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 4381 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 4441 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag 4501 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 4561 gacgagctgt acaagggccg gccttaaggc ctcgagggcc aattaattaa ctctagataa 4621 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 4681 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtcgag 4861 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4921 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4981 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5041 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5101 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5161 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5221 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5521 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 5581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5761 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 5821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7081 catttccccg aaaagtgcca cctgacgtc //