LOCUS Exported 6645 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pRS416 carrying Gateway cassette for a single insert. ACCESSION . VERSION . KEYWORDS p416 1-F GW SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6645) TITLE Fluorescent X-QTL JOURNAL Unpublished REFERENCE 2 (bases 1 to 6645) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6645 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 70..573 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 610..714 /gene="bla" /label=AmpR promoter CDS 715..1575 /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(933..952) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1746..2334 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2235..2254 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2488..2505 /label=L4440 /note="L4440 vector, forward primer" promoter 2658..2688 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2696..2712 /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 2701..2723 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2720..2736 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2720..2736 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2755..2775 /label=T3 /note="T3 promoter, forward primer" promoter 2757..2775 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind 2812..2828 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 2812..2828 /label=pBluescriptSK /note="For pBluescript vector" protein_bind 2853..2977 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 3002..3032 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 3086..3745 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(3152..3171) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 4090..4395 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 4274..4293 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(4436..4560) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4609..4625) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4610..4626) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(4650..4669) /label=T7 /note="T7 promoter, forward primer" promoter complement(4651..4669) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4679..4696) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4679..4695) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4688..4710) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4840..5295 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4927..4946) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5137..5158 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(5426..6229) /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 complement(6230..6445) /gene="S. cerevisiae URA3" /label=URA3 promoter primer_bind complement(6476..6495) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6595..6617 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata 121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt 181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa 241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata 301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct 361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata 421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact 481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa 541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg 601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact 2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg 2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt 2761 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaact 2821 agtggatccc ccgaactagt ggatccccca tcacaagttt gtacaaaaaa gctgaacgag 2881 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta 2941 cataatactg taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg 3001 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag 3061 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga 3121 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac 3181 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa 3241 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga 3301 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta 3361 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga 3421 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc 3481 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag 3541 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac 3601 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca 3661 tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg 3721 cgatgagtgg cagggcgggg cgtaaacgcc gcgtggatcc ggcttactaa aagccagata 3781 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac 3841 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc 3901 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa 3961 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag 4021 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg 4081 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt 4141 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc 4201 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg 4261 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg 4321 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg 4381 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag 4441 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt 4501 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt 4561 gatgggctgc aggaattcgg ggctgcagga attcgatatc aagcttatcg ataccgtcga 4621 cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac 4681 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc 4741 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc 4801 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat 4861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 4921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 4981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 5041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 5101 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 5161 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 5221 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 5281 taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca 5341 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac 5401 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt 5461 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata 5521 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat 5581 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc 5641 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat 5701 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc 5761 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg 5821 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt 5881 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa 5941 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta 6001 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta 6061 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat 6121 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa 6181 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc 6241 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt 6301 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt 6361 ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag 6421 aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt atggtgcact ctcagtacaa 6481 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc 6541 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga 6601 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga //