LOCUS Exported 8711 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pYX142-mtGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8711) 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 8711) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8711 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(259..287) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 882..902 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" CDS 967..993 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(1121..1140) /label=T7 /note="T7 promoter, forward primer" promoter complement(1122..1140) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1147..1164) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1147..1163) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1156..1178) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 1304..1759 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1391..1410) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1601..1622 /label=F1ori-F /note="F1 origin, forward primer" promoter 2060..2467 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 2468..3562 /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSATMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" primer_bind complement(4056..4075) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4175..4197 /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 4295..4798 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 4835..4939 /gene="bla" /label=AmpR promoter CDS 4940..5800 /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" rep_origin 5971..6559 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6460..6479 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6713..6730 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6847..6868 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6883..6913 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6921..6937 /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 6926..6948 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6945..6961 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6945..6961 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 6982..7000 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter complement(7581..7685) /gene="bla" /label=AmpR promoter ORIGIN 1 atggcctcca ctcgtgtcct cgcctctcgc ctggcctccc ggatggctgc ttccgccaag 61 gttgcccgcc ctgctgtccg cgttgctcag gtcagcaagc gcaccatcca gactggctcc 121 cccctccaga ccctcaagcg cacccagatg acctccatcg tcaacgccac cacccgccag 181 gctttccaga agcgcgccta ctcttccgga tccccgggta ccagatctat gagtaagggt 241 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg 301 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt 361 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc 421 acttatggtg ttcaatgctt ttcaagatac ccagatcata tgaaacggca tgactttttc 481 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg 541 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag 601 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac 661 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac 721 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa 781 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa 841 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta 901 acagctgctg ggattacaca tggcatggat gaactataca aataaaagct tacgcgtcga 961 cccgggtatc cgtatgatgt gcctgactac gcatgatatc tcgagctcag ctagctaact 1021 gaataaggaa caatgaacgt ttttcctttc tcttgttcct agtattaatg actgaccgat 1081 acatcccttt ttttttttgt ctttgtctag ctccaattcg ccctatagtg agtcgtatta 1141 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact 1201 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac 1261 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc 1321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 1381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 1441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 1501 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 1561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 1621 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 1681 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 1741 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt 1801 cacaccgcat atcgaccggt cgaggagaac ttctagtata tctacatacc taatattatt 1861 gccttattaa aaatggaatc ccaacaatta catcaaaatc cacattctct tcaaaatcaa 1921 ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt atatttatag gataattata 1981 ctctatttct caacaagtaa ttggttgttt ggccgagcgg tctaaggcgc ctgattcaag 2041 aaatatcttg accgcagtta actgtgggaa tactcaggta tcgtaagatg caagagttcg 2101 aatctcttag caaccattat ttttttcctc aacataacga gaacacacag gggcgctatc 2161 gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc gcctgacgca 2221 tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagagcgc cggaaccggc 2281 ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat acttgaagtt 2341 gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat cgtcttactt 2401 tctaactttt cttacctttt acatttcagc aatatatata tatatatttc aaggatatac 2461 cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga 2521 aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa 2581 gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgttccact 2641 tccagatgag gcgctggaag cctccaagaa ggctgatgcc gttttgttag gtgctgtggg 2701 tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa 2761 agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga 2821 cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt 2881 agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga 2941 tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc 3001 cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc 3061 ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgagttcc ctacattgaa 3121 ggttcaacat caattgattg attctgccac catgatccta gttaagaacc caacccacct 3181 aaatggtatt ataatcacca gcaacatgtt tggtgacatc atctccgatg aagcctccgt 3241 tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa 3301 caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa 3361 ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt 3421 gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag 3481 aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga 3541 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt 3601 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg 3661 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt 3721 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg 3781 cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga 3841 aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa aaaaaaaaaa 3901 atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa gtcaatacct 3961 tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg 4021 gccttaacga cgtagtcgat atggtgcact ctcagtacaa tctgctctga tgccgcatag 4081 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc 4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt 4201 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag 4261 gttaatgtca tgataataat ggtttcttag gacggatcgc ttgcctgtaa cttacacgcg 4321 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat 4381 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa 4441 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta 4501 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa 4561 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat 4621 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct 4681 tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt 4741 ttaatttata tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga 4801 aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 4861 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 4921 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 4981 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 5041 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 5101 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 5161 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 5221 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 5281 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 5341 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 5401 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 5461 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 5521 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 5581 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 5641 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 5701 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 5761 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 5821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 5881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 5941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 6001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 6061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 6121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 6181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 6241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 6301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 6361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 6421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 6481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 6601 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 6661 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 6721 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 6781 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 6841 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 6901 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 6961 ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggtaccggg 7021 ccggccgtcg agcttgatgg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 7081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 7141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 7201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 7261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 7321 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 7381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 7441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 7501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 7561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 7621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 7681 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 7741 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt ggggatctac 7801 gtatggtcat ttcttcttca gattccctca tggagaaagt gcggcagatg tatatgacag 7861 agtcgccagt ttccaagaga ctttattcag gcacttccat gataggcaag agagaagacc 7921 cagagatgtt gttgtcctag ttacacatgg tatttattcc agagtattcc tgatgaaatg 7981 gtttagatgg acatacgaag agtttgaatc gtttaccaat gttcctaacg ggagcgtaat 8041 ggtgatggaa ctggacgaat ccatcaatag atacgtcctg aggaccgtgc tacccaaatg 8101 gactgattgt gagggagacc taactacata gtgtttaaag attacggata tttaacttac 8161 ttagaataat gccatttttt tgagttataa taatcctacg ttagtgtgag cgggatttaa 8221 actgtgagga ccttaataca ttcagacact tctgcggtat caccctactt attcccttcg 8281 agattatatc taggaaccca tcaggttggt ggaagattac ccgttctaag acttttcagc 8341 ttcctctatt gatgttacac ctggacaccc cttttctggc atccagtttt taatcttcag 8401 tggcatgtga gattctccga aattaattaa agcaatcaca caattctctc ggataccacc 8461 tcggttgaaa ctgacaggtg gtttgttacg catgctaatg caaaggagcc tatatacctt 8521 tggctcggct gctgtaacag ggaatataaa gggcagcata atttaggagt ttagtgaact 8581 tgcaacattt actattttcc cttcttacgt aaatattttt ctttttaatt ctaaatcaat 8641 ctttttcaat tttttgtttg tattcttttc ttgcttaaat ctataactac aaaaaacaca 8701 tacaggaatt c //