LOCUS Exported 5452 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast tagging vector to create gene fusion with tdTomato with SpHis5 selection. ACCESSION . VERSION . KEYWORDS pFA6a-link-tdTomato-SpHis5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5452) TITLE Yeast tagging plasmids. REFERENCE 2 (bases 1 to 5452) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5452 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 19..206 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(74..98) /label=pHybLex-R /note="ADH terminator, reverse primer" gene 281..1481 /label=HIS3MX6 /note="yeast selectable marker encoding the S. pombe his5 gene, which corresponds to S. cerevisiae HIS3" promoter 281..624 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 625..1278 /codon_start=1 /gene="Schizosaccharomyces pombe his5" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=S. pombe his5 /note="yeast auxotrophic marker; corresponds to S. cerevisiae HIS3" /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL" terminator 1284..1481 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(1585..1604) /label=T7 /note="T7 promoter, forward primer" promoter complement(1586..1604) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1691..1708) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1862..2450) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1942..1961) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2621..3481) /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 3244..3263 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3482..3586) /gene="bla" /label=AmpR promoter primer_bind 3654..3672 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3710..3732) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3832..3851 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 3932..3950 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3932..3949 /label=SP6 /note="SP6 promoter, forward primer" CDS 4020..5450 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" ORIGIN 1 cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt 61 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc 121 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc 181 gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc 241 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac 301 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat 361 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc 421 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac 481 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt 541 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca 601 catcacatcc gaacataaac aaccatgggt aggagggctt ttgtagaaag aaatacgaac 661 gaaacgaaaa tcagcgttgc catcgctttg gacaaagctc ccttacctga agagtcgaat 721 tttattgatg aacttataac ttccaagcat gcaaaccaaa agggagaaca agtaatccaa 781 gtagacacgg gaattggatt cttggatcac atgtatcatg cactggctaa acatgcaggc 841 tggagcttac gactttactc aagaggtgat ttaatcatcg atgatcatca cactgcagaa 901 gatactgcta ttgcacttgg tattgcattc aagcaggcta tgggtaactt tgccggcgtt 961 aaaagatttg gacatgctta ttgtccactt gacgaagctc tttctagaag cgtagttgac 1021 ttgtcgggac ggccctatgc tgttatcgat ttgggattaa agcgtgaaaa ggttggggaa 1081 ttgtcctgtg aaatgatccc tcacttacta tattcctttt cggtagcagc tggaattact 1141 ttgcatgtta cctgcttata tggtagtaat gaccatcatc gtgctgaaag cgcttttaaa 1201 tctctggctg ttgccatgcg cgcggctact agtcttactg gaagttctga agtcccaagc 1261 acgaagggag tgttgtaaag agtactgaca ataaaaagat tcttgttttc aagaacttgt 1321 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt 1381 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa 1441 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac 1501 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct 1561 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg 1621 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 1681 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 1741 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 1801 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 1861 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 1921 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 1981 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 2041 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 2101 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 2161 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 2221 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 2281 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 2341 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 2401 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 2461 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 2521 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 2581 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 2641 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 2701 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 2761 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 2821 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 2881 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 2941 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 3001 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 3061 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 3121 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 3181 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 3241 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 3301 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 3361 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 3421 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 3481 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 3541 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 3601 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 3661 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 3721 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 3781 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 3841 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt 3901 aatacataac cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc 3961 cagctgaagc ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca 4021 tggtgagcaa gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg 4081 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg 4141 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca 4201 tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc 4261 ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg 4321 aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct 4381 acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga 4441 ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg 4501 agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca 4561 tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg 4621 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc 4681 gccaccacct gttcctgggg catggcaccg gcagcaccgg cagcggcagc tccggcaccg 4741 cctcctccga ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca 4801 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct 4861 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct 4921 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg 4981 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga 5041 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc 5101 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga 5161 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga 5221 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca 5281 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca 5341 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg 5401 agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaagtaa gg //