LOCUS Exported 5189 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION genomic mRFP tagging in S. pombe. ACCESSION . VERSION . KEYWORDS pFA6a-mRFP-ura4MX6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5189) AUTHORS Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E TITLE New vectors for epitope tagging and gene disruption in Schizosaccharomyces pombe. JOURNAL Biotechniques. 2013 Nov;55(5):257-63. doi: 10.2144/000114100. PUBMED 24215641 REFERENCE 2 (bases 1 to 5189) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5189 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 63..740 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" primer_bind complement(193..211) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 461..480 /label=mCherry-F /note="mCherry, forward primer" primer_bind 655..678 /label=DsRed1-C /note="DsRed1, forward primer" terminator 770..957 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(825..849) /label=pHybLex-R /note="ADH terminator, reverse primer" CDS complement(1426..2220) /codon_start=1 /gene="Schizosaccharomyces pombe ura4" /product="orotidine 5'-phosphate decarboxylase, required for uracil biosynthesis" /label=S. pombe ura4 /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH" primer_bind complement(2826..2845) /label=T7 /note="T7 promoter, forward primer" promoter complement(2827..2845) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2932..2949) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3103..3691) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3183..3202) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3862..4722) /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 4485..4504 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4723..4827) /gene="bla" /label=AmpR promoter primer_bind 4895..4913 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4951..4973) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5073..5092 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 5173..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 5173..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta 61 acatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg 121 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg 181 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca 241 tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc gccgacatcc 301 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg 361 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct 421 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga 481 ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc ctgaagggcg 541 agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca 601 cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac atcaagctgg 661 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc 721 gccactccac cggcgcctaa gaattcgaag gcgcgccact tctaaataag cgaatttctt 781 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta 841 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt 901 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag 961 atccgctagg gataacaggg taatatagat ctcttgtgat attgacgaaa ctttttgaca 1021 tctaatttat tctgttccaa caccaatgtt tataaccaag ttttatcttg tttgtctaca 1081 tggtatttta cattcatcta catacatctt tcattggctt tgtacatagt tatcattaca 1141 agtctaaaaa aattcactct tttcttattc aatgtcaatc caagagaaaa gattgtggta 1201 atgttgtagg agcatgttta ataaattact atagcaaatt actttttatt cccaaggtgt 1261 ttatctataa tagttaatat tttagtcgct acataaaatt ttaccaaaga gtacttgtat 1321 actaattcta aatgccttct gacataaaac gcctaggaaa acaaacgcaa acaaggcatc 1381 gactttttca ataaccaacc aaaaaaattt tacattagtc tttttttaat gctgagaaag 1441 tctttgctga tatgccttcc aaccagcttc tctatatctc ttggcttcga caacaggatt 1501 acgaccagct ccatagactc cacgaccaac aatgatgata tcgctaccgc agtttacaat 1561 cacttcttca ggagtacgat attgctgtcc cagcccgtct cctttaacat ccaagccgat 1621 accaggggac atagttatgt agtcgctttg aaggttagga aatcgacgac cagctataaa 1681 gccaaagcaa aaatcggtat gcttctcaaa ccattctaag gttttctctg tgtaggaacc 1741 agtagccaaa gagcctttgg aagacatttc agccaaaagc aagagaccac gtcccaaagg 1801 taaaccaact tctttgaggc cttgtataat accctcgcct ggcactgtat ggcaatttgt 1861 gatatgagcc caagaagcaa ttttgtacac accagatgca tattgtagct tgacggtatt 1921 tccaatgtct gcgaatttgc gatcctcaaa gataagaaaa cgatgctttt tacctaaggc 1981 caccagtttt tctaccatat cctggtcgaa atcctcgaca acgtcaatat gtgtcttgat 2041 aacacagaca tagggtccaa ttttatctac caattctaag atttcggatt tcttcgtcaa 2101 atcgaccgcg actgacaagt tgctttgctt ttcttccatc aaagccaaca attccttggc 2161 aatgggattt ttcatcccct cagctctagc tgaatagctt tgaaatactc tagcatccat 2221 aactttgctt ttaaaccttt aatttcgatc caagcaaaaa agaggttctt ggtaggacaa 2281 tacggtaaga aaacacgaca tgtgcagaga tgccgacgaa gcatagttaa actgggatgg 2341 taaaatcaat taagaattta taaagacaaa attgtataag tctctaaaac atcttaatta 2401 tacctcacag aactatctaa aatatattca caaagtgcaa acattatcat gaaaaagaac 2461 cattttaatt taaagcaagg gcattaaggc ttatttacag aatttcttac ttttgtaaag 2521 attataaggc tgattatctt tttcaccatg ccaaaaatta cacaagatag aatggatgtt 2581 tgaaattaaa cgtgagtata caaacaaata cactaggtaa atcgaaacat ttttttctcc 2641 attaagtaac aaattcctat ttagagaaag aatgctgagt agattaaata atctatacaa 2701 acttttttaa cacaaatgca tacatatagc cagtgggatt tgtagctaag ctgtttaaac 2761 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc 2821 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat 2881 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 2941 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 3001 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 3061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 3121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 3181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 3241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg 3301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 3361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 3421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 3481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 3541 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 3601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 3661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 3721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 3781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 3841 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 3901 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 3961 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 4021 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 4081 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 4141 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 4201 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 4261 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 4321 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 4381 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 4441 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 4501 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 4561 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 4621 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 4681 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 4741 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 4801 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 4861 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 4921 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 4981 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 5041 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 5101 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat 5161 catacacata cgatttaggt gacactata //