LOCUS Exported 6839 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION yeast plasmid for integration of target sequence at HO locus, Kan selection, pGAL inducible promoter. ACCESSION . VERSION . KEYWORDS HO-pGAL-poly-KanMX4-HO SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6839) AUTHORS Voth WP, Richards JD, Shaw JM, Stillman DJ TITLE Yeast vectors for integration at the HO locus. JOURNAL Nucleic Acids Res. 2001 Jun 15;29(12):E59-9. PUBMED 11410682 REFERENCE 2 (bases 1 to 6839) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6839 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 206..224 /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind 262..278 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 263..279 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(313..329) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(313..329) /label=pBluescriptSK /note="For pBluescript vector" promoter complement(344..785) /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" primer_bind complement(350..373) /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" protein_bind 668..785 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" gene 858..2214 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 858..1201 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1202..2011 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1269..1288) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 2017..2214 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2822..2841) /label=T7 /note="T7 promoter, forward primer" promoter complement(2823..2841) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2848..2865) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2848..2864) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2857..2879) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(3164..3183) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3296..3317 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3471..3490) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3590..3612 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3650..3668) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3736..3840 /gene="bla" /label=AmpR promoter CDS 3841..4701 /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(4059..4078) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4872..5460 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5361..5380 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5614..5631 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5748..5769 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." protein_bind 5824..5840 /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 5829..5851 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5848..5864 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5848..5864 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 gtacccgagc gtcccaaaac cttctcaagc aaggttttca gtataatgtt acatgcgtac 61 acgcgtttgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac 121 ataactatta aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt 181 cggttagagc ggatgtggga ggagggcgtg aatgtaagcg tgacataact aattacatga 241 tatcgacaaa ggaaaagggg ctcgaggtcg acggtatcga taagcttgat atcgaattcc 301 tgcagcccgg gggatccact agttctagaa tccggggttt tttctccttg acgttaaagt 361 atagaggtat attaacaatt ttttgttgat acttttatta catttgaata agaagtaata 421 caaaccgaaa atgttgaaag tattagttaa agtggttatg cagtttttgc atttatatat 481 ctgttaatag atcaaaaatc atcgcttcgc tgattaatta ccccagaaat aaggctaaaa 541 aactaatcgc attatcatcc tatggttgtt aatttgattc gttcatttga aggtttgtgg 601 ggccaggtta ctgccaattt ttcctcttca taaccataaa agctagtatt gtagaatctt 661 tattgttcgg agcagtgcgg cgcgaggcac atctgcgttt caggaacgcg accggtgaag 721 acgaggacgc acggaggaga gtcttccttc ggagggctgt cacccgctcg gcggcttcta 781 atccgtacta gaggggttaa ttaaggcgcg ccagatctgt ttagcttgcc tcgtccccgc 841 cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc 901 gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat 961 aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct 1021 cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg 1081 ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata 1141 tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac 1201 catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga 1261 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg 1321 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc 1381 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc 1441 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc 1501 cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga 1561 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa 1621 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga 1681 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat 1741 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga 1801 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat 1861 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc 1921 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca 1981 gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt 2041 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat 2101 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt 2161 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg 2221 attcgatact aacgccgcca tccagtgtcg aaaacgagct cgaattaatt cctgggggaa 2281 caacttcaca gaatgttttg tcatattgtc gaagtggtca caaaacaaga gaagttccgc 2341 caattataaa aagggaaccc gtatatttca gcttcacgga tgatttccag ggtgagagta 2401 ctgtatatgg gcttacgata gaaggccata aaaatttctt gcttggcaac aaaatagaag 2461 tgaaatcatg tcgaggctgc tgtgtgggag aacagcataa aatatcacaa aaaaagaatc 2521 taaaacactg tgttgcttgt cccagaaagg gaatcaagta tttttataaa gattggagtg 2581 gtaaaaatcg agtatgtgct agatgctatg gaagatacaa attcagcggt catcactgta 2641 taaattgcaa gtatgtacca gaagcacgtg aagtgaaaaa ggcaaaagac aaaggcgaaa 2701 aattgggcat tacgcccgaa ggtttgccag ttaaaggacc agagtgtata aaatgtggcg 2761 gaatcttaca gtggcctatg cggccgctct agctagaact agtggatcga tccccaattc 2821 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga 2881 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg 2941 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 3001 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg 3061 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3121 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3181 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggtgggcca 3241 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 3301 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa 3361 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 3421 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc 3481 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga 3541 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 3601 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata 3661 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 3721 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 3781 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 3841 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 3901 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 3961 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 4021 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 4081 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 4141 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 4201 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 4261 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 4321 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 4381 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 4441 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 4501 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 4561 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 4621 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 4681 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 4741 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 4801 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 4861 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 4921 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 4981 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 5041 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 5101 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 5161 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 5221 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg 5281 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 5341 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 5401 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 5461 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 5521 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 5581 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 5641 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 5701 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 5761 ctcactcatt aggcacccca ggctttacac tttatgcttc cgcggctcgt atgttgtgtg 5821 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccacta 5881 gtccgaggcc tcgagatccg atatcgccgt ggcggccgcc agctgaagct taattatcct 5941 gggcacgagt gaaacaaagc taaaaccttt atttagcatg gccattgaat gtaacaatta 6001 tatatatcgc aagcacaaaa aatcaaggag agagaactac cactttgttc atgtgtacaa 6061 tgttcattat ctccataagc aaaaaaaaaa aaatagaaaa catatgctat aaggttgata 6121 ttctcacgag taagcggcac ttgctactta ttgacattgc agatttttgg ctacagaaat 6181 agtatattag agattataat tgctaatcaa atcaaaatat aaaattagta aaccaaacca 6241 tttataccct tccttagtag ttatggattg ttttttaatg atatttctgc aaaccaaaga 6301 aagattgtta tccagataga atttagtttt gatattcatt tttttgttga agattgaacg 6361 ccatatctgg gcctcataat tcaaaagacg gtgccattat cggtagcgtt tcgcattgta 6421 ctggatttca gaaatttcac agttgatgaa tcgaaaagaa tggtctcatt gcaacacgta 6481 aggttaagat gtcccttttt accattatag gcaataaatg aatcataaaa cgaccgtata 6541 ctggtgaaat agtagggaga acgagtacct gtagtaaaaa gtataaatca tagttaatcg 6601 ggcaatgtcc ctcgatcaag gagtattgtg tcatgttcga gacaaacgcc aacatttttg 6661 tttcttttgg acaaatgttg tttgcattta tgatccgtta tattttgatc taatgtagag 6721 ttgcacgtag ttcttactgg caaagaaatc gatgcatacc aaaaaagaat aaaggtgata 6781 tttgatcttt accgtttagt tccaacgtaa aattgtgcct ttggacttaa aatggcgtc //