LOCUS Exported 7831 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pVT100U-mtGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7831) AUTHORS Westermann B, Neupert W TITLE Mitochondria-targeted green fluorescent proteins: convenient tools for the study of organelle biogenesis in Saccharomyces cerevisiae. JOURNAL Yeast. 2000 Nov;16(15):1421-7. PUBMED 11054823 REFERENCE 2 (bases 1 to 7831) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7831 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..397 /gene="S. cerevisiae ADH1" /label=ADH1 promoter /note="promoter for alcohol dehydrogenase 1" primer_bind complement(664..692) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1287..1307 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 1545..1732 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(1600..1624) /label=pHybLex-R /note="ADH terminator, reverse primer" rep_origin 1743..3089 /label=2u ori /note="yeast 2u plasmid origin of replication" CDS complement(3763..4566) /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(4567..4778) /gene="S. cerevisiae URA3" /label=URA3 promoter rep_origin complement(5376..5889) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5571..5592) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5783..5802 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind complement(5988..6006) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6074..6178 /gene="bla" /label=AmpR promoter CDS 6179..7039 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDVGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6397..6416) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7210..7798 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7699..7718 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 caacttcttt tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca 61 atgacaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac 121 agatgtcgtt gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc 181 cttcattcac gcacactact ctctaatgag caacggtata cggccttcct tccagttact 241 tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt 301 attaatcttt tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg 361 cacaatattt caagctatac caagcataca atcaactcca agcttatggc ctccactcgt 421 gtcctcgcct ctcgcctggc ctcccggatg gctgcttccg ccaaggttgc ccgccctgct 481 gtccgcgttg ctcaggtcag caagcgcacc atccagactg gctcccccct ccagaccctc 541 aagcgcaccc agatgacctc catcgtcaac gccaccaccc gccaggcttt ccagaagcgc 601 gcctactctt ccggatcccc gggtaccaga tctatgagta agggtgaaga acttttcact 661 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc 721 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact 781 actggaaaac tacctgttcc atggccaaca cttgtcacta ctttcactta tggtgttcaa 841 tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc 901 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt 961 gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat 1021 tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat 1081 gtatacatca tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac 1141 aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc 1201 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa 1261 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt 1321 acacatggca tggatgaact atacaaataa gaattctgca gatatccatc acactggcgg 1381 ccgctcgagg agctcgtcta gaggatccag ctttggactt cttcgccaga ggtttggtca 1441 agtctccaat caaggttgtc ggcttgtcta ccttgccaga aatttacgaa aagatggaaa 1501 agggtcaaat cgttggtaga tacgttgttg acacttctaa ataagcgaat ttcttatgat 1561 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg 1621 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt 1681 tgctttctca ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgcgat 1741 atgatccaat atcaaaggaa atgatagcat tgaaggatga gactaatcca attgaggagt 1801 ggcagcatat agaacagcta aagggtagtg ctgaaggaag catacgatac cccgcatgga 1861 atgggataat atcacaggag gtactagact acctttcatc ctacataaat agacgcatat 1921 aagtacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata tatatataca 1981 ggcaacacgc agatataggt gcgacgtgaa cagtgagctg tatgtgcgca gctcgcgttg 2041 cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga agttcctatt 2101 ctctagctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgaaga 2161 cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat 2221 accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc 2281 ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac 2341 attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt 2401 catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag 2461 acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac 2521 tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt 2581 atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca 2641 ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac 2701 ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt 2761 aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa 2821 agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa 2881 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac 2941 aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct 3001 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt 3061 tctacaaaat gaagcacaga tgcttcgtta acaaagatat gctattgaag tgcaagatgg 3121 aaacgcagaa aatgaaccgg ggatgcgacg tgcaagatta cctatgcaat agatgcaata 3181 gtttctccag gaaccgaaat acatacattg tcttccgtaa agcgctagac tatatattat 3241 tatacaggtt caaatatact atctgtttca gggaaaactc ccaggttcgg atgttcaaaa 3301 ttcaatgatg ggtaacaagt acgatcgtaa atctgtaaaa cagtttgtcg gatattaggc 3361 tgtatctcct caaagcgtat tcgaatatca ttgagaagct gcattttttt tttttttttt 3421 tttttttttt tttatatata tttcaaggat ataccattgt aatgtctgcc cctaagaaga 3481 tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc 3541 ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg 3601 gtggtgctgc tatcgcagat ctggcttttt ctttccaatt tttttttttt cgtcattata 3661 aaaatcatta cgaccgagat tcccgggtaa taactgatat aattaaattg aagctctaat 3721 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca 3781 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc 3841 ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac 3901 atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc 3961 gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat 4021 aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc 4081 tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc 4141 ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc 4201 gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa 4261 tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt 4321 ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc 4381 cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc 4441 cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt 4501 aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt 4561 cgacatgatt tatcttcgtt tccggttttt gttctgtgca gttgggttaa gaatactggg 4621 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc 4681 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga 4741 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gccagatctg gatctgtttc 4801 tggctatatg ctggagtcag cactggaagc gggtctgatc atctccgatg aagcctccgt 4861 tatcccaggc tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa 4921 caccgcattt ggtttgtacg aaccatgcca tggttccgct ccagatttgc caaagaataa 4981 ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt 5041 gcctgaagaa ggtaaagcca ttgaagatgc agttaaaaag gttttggatg caggtatcag 5101 aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga 5161 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acagcgtcac 5221 atcggataat aatgatggca gccattgtag aagtgccttt tgcatttcta gtctctttct 5281 cggtctagct agttttacta catcgcgaag atagaatctt agatcacact gcctttgctg 5341 agctggatca atagagtaac aaaagagtgg taaggacccc ggttgataat cagaaaagcc 5401 ccaaaaacag gaagattgta taagcaaata tttaaattgt aagcgttaat attttgttaa 5461 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 5521 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 5581 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 5641 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc 5701 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 5761 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 5821 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 5881 agggcgcgtc ctcgttaaag gacaaggacc tgagcggaag tgtatcgtac agtagacgga 5941 gtatactagt atagtctata gtccgtggaa ttcttgaaga cgaaagggcc tcgtgatacg 6001 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 6061 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 6121 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 6181 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 6241 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 6301 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 6361 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 6421 tattgacgtc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 6481 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 6541 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 6601 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 6661 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 6721 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 6781 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 6841 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 6901 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 6961 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 7021 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 7081 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 7141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 7201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 7261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 7321 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg 7381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 7441 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 7501 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 7561 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 7621 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 7681 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 7741 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 7801 cgccagcaac gcggcccgac ctcgacgcat g //