LOCUS Exported 5849 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid used for the C-terminal tagging of Cdc14 with mCherry and the auxin-inducible degron in yeast.. ACCESSION . VERSION . KEYWORDS pG23A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5849) AUTHORS Papagiannakis A, Niebel B, Wit EC, Heinemann M TITLE Autonomous Metabolic Oscillations Robustly Gate the Early and Late Cell Cycle. JOURNAL Mol Cell. 2017 Jan 19;65(2):285-295. doi: 10.1016/j.molcel.2016.11.018. Epub 2016 Dec 15. PUBMED 27989441 REFERENCE 2 (bases 1 to 5849) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5849 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1..22 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." CDS 572..1279 /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(717..735) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 985..1004 /label=mCherry-F /note="mCherry, forward primer" terminator 1501..1690 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" gene complement(1745..2864) /label=natMX6 /note="yeast selectable marker conferring nourseothricin resistance" terminator 1745..1942 /label=TEF terminator /note="Ashbya gossypii TEF terminator" CDS complement(1948..2520) /codon_start=1 /gene="Streptomyces noursei nat1" /product="nourseothricin acetyltransferase" /label=NrsR /note="confers resistance to nourseothricin" /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT LCGLDTALYDGTASDGEQALYMSMPCP" promoter complement(2521..2864) /label=TEF promoter /note="Ashbya gossypii TEF promoter" primer_bind 3398..3419 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3573..3592) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3692..3714 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3752..3770) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3838..3942 /gene="bla" /label=AmpR promoter CDS 3943..4803 /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 complement(4161..4180) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4974..5562 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5463..5482 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5716..5733 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccgcggct 61 cgtatgttgt gtggaattaa gagatttaac catgacgccg ccatccaacg gtcatggggc 121 gcttagtgcc agaaattcga gccaaccttc tacggcgaat aatggaagta attcattcaa 181 gagctcagcc gtcccacaga cttcgcctgg tcagccgaga aagggccaaa acggttctaa 241 tactatagaa gatatcaaca ataatcgtaa tccaacttcc catgcgaata ggaaggtagt 301 aatagaaagc aacaactcag atgacgaatc catgcaggat accaacggca caagtaatca 361 ttatcccaaa gtttcgcgca agaaaaatga tatctcctct gcttcatcgt cccgaatgga 421 agataacgaa cccagcgcaa cgaacatcaa taatgccgca gatgatacca tactaagaca 481 actattgccc aagaatagga gagtaacgtc agggagaagg actacaagcg ccgccggtgg 541 tataagaaaa ataagtggct ccatcaagaa aatggtgagc aagggcgagg aggataacat 601 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca 661 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa 721 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt 781 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct 841 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt 901 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg 961 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc 1021 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct 1081 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa 1141 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa 1201 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg 1261 catggacgag ctgtacaagg tcgacggtgc aggcgccggg gctggtgctg gggcaggtac 1321 ccctaaagat ccagccaaac ctccggccaa ggcacaagtt gtgggatggc caccggtgag 1381 atcataccgg aagaacgtga tggtttcctg ccaaaaatca agcggtggcc cggaggcggc 1441 ggcgttcgtg aaggtatcac ccgggtaact cgagtaagct tggtaccgcg gctagctaag 1501 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt 1561 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct 1621 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg 1681 acgctcgaag atccagctca gcgtacgtcg acactggatg gcggcgttag tatcgaatcg 1741 acagcagtat agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac 1801 ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat 1861 ttgattaaaa tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa 1921 caagaatctt tttattgtca gtactgatta ggggcagggc atgctcatgt agagcgcctg 1981 ctcgccgtcc gaggcggtgc cgtcgtacag ggcggtgtcc aggccgcaga gggtgaaccc 2041 catccgccgg tacgcgtgga tcgccggtgc gttgacgttg gtgacctcca gccagaggtg 2101 cccggcgccc cgctcgcggg cgaactccgt cgcgagcccc atcaacgcgc gcccgacccc 2161 gtgcccccgg tgctccgggg cgacctcgat gtcctcgacg gtcagccggc ggttccagcc 2221 ggagtacgag acgaccacga agcccgccag gtcgccgtcg tccccgtacg cgacgaacgt 2281 ccgggagtcc gggtcgccgt cctccccgtc gtccgattcg tcgtccgatt cgtcgtcggg 2341 gaacaccttg gtcaggggcg ggtccaccgg cacctcccgc agggtgaagc cgtccccggt 2401 ggcggtgacg cggaagacgg tgtcggtggt gaaggaccca tccagtgcct cgatggcctc 2461 ggcgtccccc gggacactgg tgcggtaccg gtaagccgtg tcgtcaagag tggtacccat 2521 ggttgtttat gttcggatgt gatgtgagaa ctgtatccta gcaagatttt aaaaggaagt 2581 atatgaaaga agaacctcag tggcaaatcc taacctttta tatttctcta caggggcgcg 2641 gcgtggggac aattcaacgc gtctgtgagg ggagcgtttc cctgctcgca ggtctgcagc 2701 gaggagccgt aatttttgct tcgcgccgtg cggccatcaa aatgtatgga tgcaaatgat 2761 tatacatggg gatgtatggg ctaaatgtac gggcgacagt cacatcatgc ccctgagctg 2821 cgcacgtcaa gactgtcaag gagggtattc tgggcctcca tgtcgctggc cgggtgaccc 2881 ggcggggaca aggcaagctc agtgtccctt ctcgctgtct tttcgaaaga gtcatatgta 2941 taccggcaat agtaacttgc ctgatttatg cataatgccg ttgtatttgc tagccagttt 3001 tccaccgctg tctgtgggtt gcgaccatgc tacctggtac ctaggaataa cttcaccgta 3061 aacaatattt tctcttgcaa cgtcaatcaa tttggttgcg atacccaact tccttgctgt 3121 cctgcacacc caaattctcg atatgccaat cttaaaatcg gggtacacat tctgtaccaa 3181 tcttctagaa tcataaacca tccaacgtcc acgactagat gttttaccat tgcccccata 3241 aaggttctct ataattatta ttccgaccgc cctgtcattt cttatgtata caaacgcttt 3301 gcctttttct tctgtggtac tgttccaaat gacattctca tcgtgtggcg cattcagctc 3361 gttattcact agtgtcatta tctccgtcat ggtaatagtg gactcttgtt ccaaactgga 3421 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg 3481 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 3541 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 3601 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 3661 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 3721 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 3781 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 3841 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 3901 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 3961 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 4021 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 4081 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 4141 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 4201 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 4261 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 4321 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 4381 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 4441 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 4501 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 4561 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 4621 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 4681 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 4741 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 4801 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 4861 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 4921 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 4981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 5041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 5101 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 5161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 5221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcacag 5281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 5341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 5401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 5461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 5521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 5581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 5641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 5701 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 5761 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac 5821 tggaaagcgg gcagtgagcg caacgcaat //