LOCUS Exported 6272 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synchronize trafficking of ManII from the ER (RUSH system). ACCESSION . VERSION . KEYWORDS Str-KDEL_ManII-SBP-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6272) 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 6272) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6272 /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 2908..3021 /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 3031..3738 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(3176..3194) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3444..3463 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3892..4013 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3929..3948) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4115..4131) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4115..4131) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4128..4150) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4139..4155 /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(4163..4193) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4208..4229 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4346..4363) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4517..5105) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4597..4616) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5276..6136) /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 5899..5918 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6137..6241) /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 ccatgaagtt aagtcgccag ttcaccgtgt ttggcagcgc gatcttctgc 2581 gtcgtaatct tctcactcta cctgatgctg gacaggggtc acttggacta ccctcggggc 2641 ccgcgccagg agggctcctt tccgcagggc cagctttcaa tattgcaaga aaagattgac 2701 catttggagc gtttgctcgc tgagaacaac gagattatct caaatatcag agactcagtc 2761 atcaacctga gcgagtctgt ggaggacggc ccgcgggggt caccaggcaa cgccagccaa 2821 ggctccatcc acctccactc gccacagttg gccctgcagg ctgaccccag agactgtttg 2881 gatcccaccg gtcgccaccg gaattccatg gacgagaaga ccactggttg gcgaggtgga 2941 cacgttgttg aaggactggc tggggaactt gaacaacttc gtgcacgact ggagcatcac 3001 ccacaaggtc aacgtgaacc acctgcaggt atggtgagca agggcgagga ggataacatg 3061 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac 3121 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag 3181 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc 3241 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg 3301 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg 3361 accgtgaccc aggactcctc cctacaggac ggcgagttca tctacaaggt gaagctgcgc 3421 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc 3481 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg 3541 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag 3601 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac 3661 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc 3721 atggacgagc tgtacaaggg ccggccttaa ggcctcgagg gccaattaat taactctaga 3781 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 3841 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 3901 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 3961 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc 4021 gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 4081 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 4141 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 4201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 4261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 4321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 4381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 4441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 4501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 4561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 4621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 4681 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg 4741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 4801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 4861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 4921 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc 4981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 5101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 5161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 5221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 5281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 5401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 5461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 5521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 5581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 5641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 5701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 5761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 5821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 5881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 5941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 6001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 6061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 6181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 6241 gcacatttcc ccgaaaagtg ccacctgacg tc //