LOCUS Exported 8831 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDR-GW AT1.03YEMK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8831) AUTHORS Bermejo C, Haerizadeh F, Takanaga H, Chermak D, Frommer WB TITLE Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors. JOURNAL Biochem J. 2010 Dec 1. 432(2):399-406. PUBMED 20854260 REFERENCE 2 (bases 1 to 8831) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8831 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 743..760 /label=L4440 /note="L4440 vector, forward primer" protein_bind 877..898 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 913..943 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 951..967 /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)." primer_bind 956..978 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 975..991 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 975..991 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" rep_origin complement(1021..1476) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1158..1179) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1370..1389 /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2284..2308 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3433..4167 /codon_start=1 /product="circularly permuted form of the Venus yellow fluorescent protein (Nagai et al., 2004)" /label=cp173Venus /note="efficient FRET acceptor for CFP" /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRD HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN VYITADKQKNGIKANFKIRHNIE" protein_bind complement(4205..4229) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(4291..4310) /label=T7 /note="T7 promoter, forward primer" promoter complement(4292..4310) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator 4458..4645 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(4513..4537) /label=pHybLex-R /note="ADH terminator, reverse primer" primer_bind complement(4657..4674) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4657..4673) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4666..4688) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5109..6455 /label=2u ori /note="yeast 2u plasmid origin of replication" CDS complement(6566..7369) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(7370..7585) /gene="S. cerevisiae URA3" /label=URA3 promoter primer_bind complement(7610..7628) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7696..7800 /gene="bla" /label=AmpR promoter CDS 7801..8661 /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(8019..8038) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctta ccgcatcagg 1021 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 1081 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 1141 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 1201 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 1261 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 1321 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 1381 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 1441 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccaagct tcctgaaacg 1501 gagaaacata aacaggcatt gctgggatca cccatacatc actctgtttt gcctgacctt 1561 ttccggtaat ttgaaaacaa acccggtctc gaagcggaga tccggcgata attaccgcag 1621 aaataaaccc atacacgaga cgtagaacca gccgcacatg gccggagaaa ctcctgcgag 1681 aatttcgtaa actcgcgcgc attgcatctg tatttcctaa tgcggcactt ccaggcctcg 1741 atcgagaccg tttatccatt gcttttttgt tgtctttttc cctcgttcac agaaagtctg 1801 aagaagctat agtagaacta tgagcttttt ttgtttctgt tttccttttt ttttttttta 1861 cctctgtgga aattgttact ctcacactct ttagttcgtt tgtttgtttt gtttattcca 1921 attatgaccg gtgacgaaac gtggtcgatg gtgggtaccg cttatgctcc cctccattag 1981 tttcgattat ataaaaaggc caaatattgt attattttca aatgtcctat cattatcgtc 2041 taacatctaa tttctcttaa attttttctc tttctttcct ataacaccaa tagtgaaaat 2101 ctttttttct tctatatcta caaaaacttt ttttttctat caacctcgtt gataaatttt 2161 ttctttaaca atcgttaata attaattaat tggaaaataa ccattttttc tctcttttat 2221 acacacattc aaaagaaaga aaaaaaatat accccagcct cgactagtga attcgcggcc 2281 atcacaagtt tgtacaaaaa agcaggcttc gtttaaacgg gccctctaga ctcgagtatg 2341 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 2401 gacgtaaacg gccacaggtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 2461 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 2521 gtgaccaccc tgacctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag 2581 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgtac catcttcttc 2641 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 2701 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 2761 ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc 2821 atcaaggccc acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac 2881 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 2941 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg 3001 ctggagttcg tgaccgccgc catcgatatg aaaactgtga aagtgaatat aacaacccct 3061 gatgggccag tctacgacgc tgatatcgag atggtgtccg tgcgggccga gagtggtgat 3121 ctcggcatcc tccccggtca cattcccaca aaggccccac tgaagatcgg agctgtgcgg 3181 ctgaagaagg acggccaaac cgagtacgtc gcagtctcag gcggcactgt tgaagtgcgg 3241 cctgaccacg ttaccattaa tgctcaagcc gctgaaacag ccgaaggaat cgacaaagag 3301 agagcagaag ccgcaagaca gagggcccag gagcggctga actctcaatc cgatgacacc 3361 gatattcgcc gggccgagct ggcactgcag agggccctga acagactgga cgtggctgaa 3421 atgaaggaat tcatggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc 3481 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg 3541 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 3601 gggatcactc tcggcatgga cgagctgtac aagggtggca gcggtggcat ggtgagcaag 3661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 3721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 3781 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 3841 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc 3901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 3961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 4021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 4081 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc 4141 aacttcaaga tccgccacaa catcgagtaa gcttaagttt aaaccgctga tcagcctcga 4201 ctgcacccag ctttcttgta caaagtggtg tagttaattc atgatggccg ctgcaggtcg 4261 acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcggatc 4321 cagctttgga cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt 4381 ctaccttgcc agaaatttac gaaaagatgg aaaagggtca aatcgttggt agatacgttg 4441 ttgacacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta 4501 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct 4561 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg 4621 ctcttattga ccacacctct accggcatgc caattcactg gccgtcgttt tacaacgtcg 4681 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 4741 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 4801 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca 4861 ccgcataatc ggatcgtact tgttacccat cattgaattt tgaacatccg aacctgggag 4921 ttttccctga aacagatagt atatttgaac ctgtataata atatatagtc tagcgcttta 4981 cggaagacaa tgtatgtatt tcggttcctg gagaaactat tgcatctatt gcataggtaa 5041 tcttgcacgt cgcatccccg gttcattttc tgcgtttcca tcttgcactt caatagcata 5101 tctttgttaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc 5161 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag 5221 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag 5281 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc 5341 gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc 5401 ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct 5461 ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc 5521 tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact 5581 gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct 5641 ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca 5701 ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa 5761 tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt 5821 tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa 5881 gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag 5941 caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg 6001 ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc 6061 aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcg gaataggaac 6121 ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag 6181 ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag 6241 aacggcatag tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat 6301 gaaaggtagt ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt 6361 ccttcagcac taccctttag ctgttctata tgctgccact cctcaattgg attagtctca 6421 tccttcaatg ctatcatttc ctttgatatt ggatcgatcc gatgataagc tgtcaaacat 6481 gagaattggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac 6541 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt 6601 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata 6661 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat 6721 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc 6781 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat 6841 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc 6901 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg 6961 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt 7021 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa 7081 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta 7141 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta 7201 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat 7261 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa 7321 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc 7381 gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt 7441 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt 7501 ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag 7561 aataaaaaaa aaatgatgaa ttgaaaagct aattcttgaa gacgaaaggg cctcgtgata 7621 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 7681 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 7741 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 7801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 7861 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 7921 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 7981 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 8041 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 8101 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 8161 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 8221 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 8281 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 8341 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 8401 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 8461 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 8521 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 8581 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 8641 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 8701 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 8761 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 8821 aaaggatctt c //