LOCUS Exported 6641 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synchronize trafficking of TNF from the ER (RUSH system). ACCESSION . VERSION . KEYWORDS Str-KDEL_TNF-SBP-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6641) 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 6641) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6641 /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 3277..3390 /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 3400..4107 /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(3545..3563) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3813..3832 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 4261..4382 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4298..4317) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4484..4500) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4484..4500) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4497..4519) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4508..4524 /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(4532..4562) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4577..4598 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4715..4732) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4886..5474) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4966..4985) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5645..6505) /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 6268..6287 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6506..6610) /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 ccatgtctac cgagtctatg attagggacg tggaactggc tgaggaggca 2581 ctgcccaaaa aaaccggcgg accacagggc tctaggagat gtctgtttct gtctctgttc 2641 tcttttctca tcgtggctgg cgctacaaca ctcttctgtc tgctccattt cggcgtgatt 2701 ggaccacagc gagaggaatt tccccgggat ctgtcactca tttcaccact ggcacaggct 2761 gtccgatctt catctcggac tccatccgac aaacctgtcg cccatgtcgt cgccaaccca 2821 caggccgagg gccagctcca gtggctcaat aggagggcaa acgctctgct cgccaatggc 2881 gtggaactcc gggataacca gctcgtcgtg cctagtgagg gactgtacct catctactcc 2941 caggtgctgt ttaagggcca gggatgtcct tctacacatg tgctgctcac acacacaatt 3001 tcacggatcg ccgtgtctta ccagactaaa gtcaatctgc tctctgccat caaatcccca 3061 tgtcagcggg aaacacctga gggcgctgag gctaaacctt ggtacgaacc catctacctc 3121 ggaggcgtgt tccagctgga gaagggcgat agactgagtg ctgagatcaa tcgacccgac 3181 tacctcgact ttgccgaatc tggccaggtc tactttggca tcattgctct gggcctcgag 3241 ggccgagctc atggcgcacc taggcctttg aattccatgg acgagaagac cactggttgg 3301 cgaggtggac acgttgttga aggactggct ggggaacttg aacaacttcg tgcacgactg 3361 gagcatcacc cacaaggtca acgtgaacca cctgcaggta tggtgagcaa gggcgaggag 3421 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg 3481 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag 3541 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc 3601 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac 3661 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc 3721 ggcgtggtga ccgtgaccca ggactcctcc ctacaggacg gcgagttcat ctacaaggtg 3781 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc 3841 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag 3901 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag 3961 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc 4021 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc 4081 accggcggca tggacgagct gtacaagggc cggccttaag gcctcgaggg ccaattaatt 4141 aactctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta 4201 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 4261 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4321 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4381 taacgcgtcg agtgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 4441 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 4501 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 4561 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 4621 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 4681 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 4741 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 4801 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 4861 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 4921 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 4981 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5041 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc 5101 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 5161 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 5221 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 5281 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 5341 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 5401 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 5461 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 5521 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 5581 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 5641 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 5701 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 5761 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 5821 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 5881 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 5941 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6001 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6061 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 6121 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 6181 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6241 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 6301 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 6361 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 6421 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 6481 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 6541 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 6601 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //