LOCUS Exported 6379 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 6379) 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 6379) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6379 /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" regulatory 900..909 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1065..1835 /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 1842..1883 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" promoter complement(1931..1949) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1932..1949) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1969..1986) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1975..2199 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2245..2673 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2332..2351) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2542..2563 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2682..2702) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2687..3017 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2868..3003 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2930..2949 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3084..3878 /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(3138..3157) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3748..3767 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4052..4173 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4089..4108) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4143..4162 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4222..4238) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4222..4238) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4235..4257) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4246..4262 /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(4270..4300) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4315..4336 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4453..4470) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4624..5212) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4704..4723) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5383..6243) /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 6006..6025 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6244..6348) /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 gcttgggccg 901 ccaccatgga ccctgtggtc gtgctggggc tgtgcctgag ctgtctgctg ctgctgagcc 961 tgtggaagca gtcctacgga ggcgggggat caggaagcgg cggccgcgga tcaggaagcg 1021 gatcaggaag cggatcagga agcggatcag gaagcggatc aggaatcccg ctgctgaacg 1081 ctaaacagat tctgggacag ctggacggcg ggagcgtggc agtcctgcct gtggtcgact 1141 ccaccaatca gtacctgctg gatcgaatcg gcgagctgaa gagtggggat gcttgcattg 1201 cagaatatca gcaggcaggg agaggaagca gagggaggaa atggttctct ccttttggag 1261 ctaacctgta cctgagtatg ttttggcgcc tgaagcgggg accagcagca atcggcctgg 1321 gcccggtcat cggaattgtc atggcagaag cgctgcgaaa gctgggagca gacaaggtgc 1381 gagtcaaatg gcccaatgac ctgtatctgc aggatagaaa gctggcaggc atcctggtgg 1441 agctggccgg aataacaggc gatgctgcac agatcgtcat tggcgccggg attaacgtgg 1501 ctatgaggcg cgtggaggaa agcgtggtca atcagggctg gatcacactg caggaagcag 1561 ggattaacct ggacaggaat actctggccg ctatgctgat ccgagagctg cgggcagccc 1621 tggaactgtt cgagcaggaa ggcctggctc catatctgtc acggtgggag aagctggata 1681 acttcatcaa tagacccgtg aagctgatca ttggggacaa agagattttc gggattagcc 1741 gggggattga taaacaggga gccctgctgc tggaacagga cggagttatc aaaccctgga 1801 tgggcggaga aatcagtctg cggtctgccg aaaaggctag cggcaagccc atccccaacc 1861 ccctgctggg cctggacagc acctaatgag gatccggcaa gcccatcccc aaccccctgc 1921 tgggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc 1981 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 2041 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 2101 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 2161 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca 2221 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg 2281 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 2341 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 2401 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 2461 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 2521 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 2581 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa 2641 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg 2701 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt 2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 2941 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3001 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 3061 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 3121 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 3181 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 3241 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 3301 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 3361 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 3421 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 3481 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 3541 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 3601 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 3661 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 3721 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 3781 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 3841 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 3901 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 3961 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 4021 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 4081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 4141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 4201 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4801 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5041 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 5101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 5221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 5941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 6061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 6121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 6181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6361 aaaagtgcca cctgacgtc //