LOCUS Exported 6378 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses V5-tagged miniTurbo on the mammalian ER membrane. ACCESSION . VERSION . KEYWORDS C1(1-29)-miniTurbo-V5_pCDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6378) AUTHORS Branon TC, Bosch JA, Sanchez AD, Udeshi ND, Svinkina T, Carr SA, Feldman JL, Perrimon N, Ting AY TITLE Efficient proximity labeling in living cells and organisms with TurboID. JOURNAL Nat Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201. PUBMED 30125270 REFERENCE 2 (bases 1 to 6378) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6378 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 169..548 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 549..752 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 703..723 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 797..816 /label=T7 /note="T7 promoter, forward primer" promoter 797..815 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 834..843 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 999..1769 /codon_start=1 /product="N-terminally truncated mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" /label=miniTurbo /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" CDS 1776..1817 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" promoter complement(1865..1883) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1866..1883) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1903..1920) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1909..2133 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2179..2607 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2266..2285) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2476..2497 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2616..2636) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2621..2950 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2801..2936 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2863..2882 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3017..3811 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3071..3090) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3681..3700 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3985..4106 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4022..4041) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4076..4095 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4155..4171) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4155..4171) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4168..4190) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4179..4195 /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(4203..4233) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4248..4269 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4386..4403) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4557..5145) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4637..4656) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5316..6176) /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 5939..5958 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6177..6281) /gene="bla" /label=AmpR promoter primer_bind complement(6356..6375) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca 61 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg 121 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat 181 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt 241 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 301 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac 361 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 421 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc 481 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 541 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac 601 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc 661 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc 721 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt 781 actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg gccgccacca 841 tggaccctgt ggtcgtgctg gggctgtgcc tgagctgtct gctgctgctg agcctgtgga 901 agcagtccta cggaggcggg ggatcaggaa gcggcggccg cggatcagga agcggatcag 961 gaagcggatc aggaagcgga tcaggaagcg gatcaggaat cccgctgctg aacgctaaac 1021 agattctggg acagctggac ggcgggagcg tggcagtcct gcctgtggtc gactccacca 1081 atcagtacct gctggatcga atcggcgagc tgaagagtgg ggatgcttgc attgcagaat 1141 atcagcaggc agggagagga agcagaggga ggaaatggtt ctctcctttt ggagctaacc 1201 tgtacctgag tatgttttgg cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg 1261 tcatcggaat tgtcatggca gaagcgctgc gaaagctggg agcagacaag gtgcgagtca 1321 aatggcccaa tgacctgtat ctgcaggata gaaagctggc aggcatcctg gtggagctgg 1381 ccggaataac aggcgatgct gcacagatcg tcattggcgc cgggattaac gtggctatga 1441 ggcgcgtgga ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa gcagggatta 1501 acctggacag gaatactctg gccgctatgc tgatccgaga gctgcgggca gccctggaac 1561 tgttcgagca ggaaggcctg gctccatatc tgtcacggtg ggagaagctg gataacttca 1621 tcaatagacc cgtgaagctg atcattgggg acaaagagat tttcgggatt agccggggga 1681 ttgataaaca gggagccctg ctgctggaac aggacggagt tatcaaaccc tggatgggcg 1741 gagaaatcag tctgcggtct gccgaaaagg ctagcggcaa gcccatcccc aaccccctgc 1801 tgggcctgga cagcacctaa tgaggatccg gcaagcccat ccccaacccc ctgctgggcc 1861 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta 1921 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca 1981 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc 2041 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata 2101 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg 2161 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 2221 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 2281 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 2341 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 2401 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 2461 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 2521 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 2581 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg 2641 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 2701 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 2761 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 2821 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 2881 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 2941 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg 3001 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 3061 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 3121 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 3181 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 3241 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 3301 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 3361 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 3421 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 3481 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 3541 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 3601 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 3661 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 3721 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 3781 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 3841 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 3901 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 3961 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 4021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4081 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 4141 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 4201 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 4261 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 4321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 4501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 4561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 4621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 4681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 4741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 4801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 4861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 4921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 4981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 5101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 5161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 5221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 5281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 5341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 5401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 5461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 5521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 5581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 5641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 5701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 5761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 5821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 5881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 5941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 6061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 6121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 6181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 6241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6301 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa 6361 tctgctctga tgccgcat //