LOCUS Exported 6693 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pHIS3p:mTurquoise2-Tub1+3'UTR::URA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6693) AUTHORS Markus SM, Omer S, Baranowski K, Lee WL TITLE Improved Plasmids for Fluorescent Protein Tagging of Microtubules in Saccharomyces cerevisiae. JOURNAL Traffic. 2015 Jul;16(7):773-86. doi: 10.1111/tra.12276. Epub 2015 Apr 28. PUBMED 25711127 REFERENCE 2 (bases 1 to 6693) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6693 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 72..94 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(132..150) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 218..322 /gene="bla" /label=AmpR promoter CDS 323..1183 /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(541..560) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1354..1942 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1843..1862 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2096..2113 /label=L4440 /note="L4440 vector, forward primer" primer_bind 2200..2219 /label=T7 /note="T7 promoter, forward primer" promoter 2200..2218 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 2306..2521 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 2522..3325 /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 3518..3704 /gene="S. cerevisiae HIS3" /label=HIS3 promoter CDS 3706..4422 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3751..3772) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4012..4031) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4359..4380 /label=EGFP-C /note="EGFP, forward primer" RBS 5907..5915 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" promoter complement(6547..6565) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(6548..6565) /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc 61 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc 121 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt 181 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 241 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 301 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 361 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 421 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 481 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 541 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 601 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 661 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 721 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 781 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 841 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 901 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 961 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 1021 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 1081 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 1141 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 1201 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 1261 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 1321 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 1381 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 1441 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 1501 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 1561 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 1621 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 1681 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 1741 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 1801 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 1861 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 1921 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 1981 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 2041 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 2101 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 2161 gccgattcat taatgcaggt taacctggct tatcgaaatt aatacgactc actataggga 2221 gaccggcaga tccgcggccg cataggccac tagttaacta tgcggcatca gagcagattg 2281 tactgagagt gcaccatacc accttttcaa ttcatcattt tttttttatt cttttttttg 2341 atttcggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag 2401 gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa 2461 ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac gaagataaat 2521 catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa 2581 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac 2641 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac 2701 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt 2761 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac 2821 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa 2881 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt 2941 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc 3001 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat 3061 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat 3121 gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt atagaaccgt 3181 ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa 3241 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt 3301 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa 3361 ctcacaaatt agagcttcaa tttaattata tcagttatta ccctatgcgg tgtgaaatac 3421 cgcacagatg cgtaaggaga aaataccgca tcaggggcgc gccttaatta accactaaag 3481 ggaacaaaag ctggtaccct attactcttg gcctcctcta gtacactcta tattttttta 3541 tgcctcggta atgattttca tttttttttt ccacctagcg gatgactctt tttttttctt 3601 agcgattggc attatcacat aatgaattat acattatata aagtaatgtg atttcttcga 3661 agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagcatggt gagcaagggc 3721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 3781 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 3841 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 3901 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 3961 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 4021 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 4081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 4141 tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac 4201 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag 4261 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 4321 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 4381 accgccgccg ggatcactct cggcatggac gagctgtaca agggatccat gagagaagtt 4441 attagtatta atggtatgtt cgatttgccc gtccaggcta gatctttttt ttgttagttc 4501 attttggcgt ttcatctttc attactaact tcaacgcaaa attttttttt ttttgatttc 4561 tctttacagt cggtcaagct ggttgtcaga ttggtaatgc ctgttgggaa ttatattccc 4621 ttgagcacgg tattaagccg gatggacatc tagaagatgg cctttcaaag ccgaagggag 4681 gagaagaggg tttctccacg tttttccatg aaaccggcta cggtaagttc gttccaaggg 4741 ctatttacgt ggatttagag cccaacgtta ttgacgaagt ccgtaacggt ccttacaagg 4801 acttgttcca tccagaacag ttgatcagtg gtaaggagga cgcggctaat aattatgcaa 4861 gaggccatta caccgttggt agagaaattt tgggcgatgt tctggatagg attagaaaac 4921 tggcagacca atgtgatggg ttacaagggt tcttgtttac ccattctctt ggtggtggta 4981 ctggttccgg tctagggtcc ttactgttgg aagaactatc cgctgaatac ggtaagaaat 5041 ccaagctgga atttgccgta taccctgctc cacaagtgtc tacttctgtg gttgagcctt 5101 acaacacagt tttaactaca catactacat tggaacatgc agattgtact ttcatggtcg 5161 ataatgaggc tatttacgac atgtgcaaaa gaaacttgga tatcccaaga ccaagctttg 5221 caaacttaaa caacctaatt gctcaagtgg tatcatctgt tacagcatca ttgagattcg 5281 acggttcatt aaatgtagat ttgaacgaat ttcaaaccaa tttggttcca tatccaagaa 5341 ttcatttccc cttagtttca tattctccag tcttatccaa atcaaaggca ttccatgagt 5401 ccaactctgt gtcagaaatt acaaacgctt gttttgaacc tggtaaccag atggtcaagt 5461 gtgatccaag agatggtaaa tacatggcta cttgtctgtt atacaggggt gatgtggtaa 5521 caagagatgt tcaaagagct gtcgagcagg tgaaaaacaa gaagaccgtc caattggttg 5581 attggtgtcc aactggtttc aagatcggta tttgctacga acctccaact gccacaccaa 5641 actcacaatt ggccactgtg gatagggccg tctgtatgtt gtcaaatacc acatccattg 5701 ctgaggcttg gaagagaatc gatagaaaat tcgatttaat gtatgccaaa cgtgctttcg 5761 tccactggta tgtcggtgaa ggtatggaag aaggtgaatt caccgaagct agagaagatt 5821 tggctgcttt agaaagagat tacatcgaag tgggtgccga ctcatacgct gaggaagagg 5881 aattttaagg ttcactctcg cccccaacct ccttatcctt tctatatttc aagtatattg 5941 taaaaaaaaa aattccccct actgttatat ctctttggtg atatcttatc tataatctac 6001 ttattatatt tctctaatat ctatgtacaa aaaataaaaa aaaaaaccat tctccttctc 6061 taacctgcat ttgatgaaag cttagtaagt ctcatcgtag ttgatgggcc aaagacattt 6121 gtacttgttt catcagattc ttacatcacc catttccatg attatttcac ggatttcatt 6181 agtaattgca tttttgtatc cctagttcct tttctcccgc cttgccgtcg aattgaaaaa 6241 aaaaatggaa atgaggacat cttccattga tatccaggac aaagcaagag tagaagtaac 6301 tattagcaag gagaattcca aaaatagtga acttcaaggt aagcagatct atagtttcaa 6361 tatctgattt tctgcttttc aaaacttaaa attcattaga aggtaaaaca agaacccgta 6421 aaattaaagt taccgatatt atcaggtttt tcaccccatg tctactatcc catttagaaa 6481 gaactatgtg ttcaaaaact gggccggtcg acctgcagcg tacgaagctt cagctggcgg 6541 ccgcgttcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt 6601 gtagccgcgt tctaacgaca atatgtccat atggtgcact ctcagtacaa tctgctctga 6661 tgccgcatag ttaagccagc cccgacaccc gcc //