LOCUS Exported 7084 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pRS416 carrying Gateway cassette for three inserts. ACCESSION . VERSION . KEYWORDS p416 3-F GW SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7084) TITLE Fluorescent X-QTL JOURNAL Unpublished REFERENCE 2 (bases 1 to 7084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7084 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 201..416 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 417..1220 /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" rep_origin complement(1351..1806) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1488..1509) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1700..1719 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1936..1958 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1950..1967 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1951..1967 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1977..1996 /label=T7 /note="T7 promoter, forward primer" promoter 1977..1995 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 2020..2036 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2021..2037 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 2085..2208 /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 2354..2456 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 2457..3110 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG" primer_bind complement(2523..2542) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 3460..3765 /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 3644..3663 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(4113..4237) /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4257..4273) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4257..4273) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(4310..4330) /label=T3 /note="T3 promoter, forward primer" promoter complement(4310..4328) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4349..4365) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4349..4365) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4362..4384) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4373..4389 /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)." promoter complement(4397..4427) /label=lac promoter /note="promoter for the E. coli lac operon" primer_bind complement(4580..4597) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4751..5339) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4831..4850) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5510..6370) /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 6133..6152 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6371..6475) /gene="bla" /label=AmpR promoter misc_feature 6512..7015 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind 7057..7075 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc 241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg 301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc 361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt 421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat 481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca 541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg 601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg 661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca 721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac 781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa 841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg 901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct 961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac 1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg 1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa 1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa 1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac 1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac 1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat 1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg 1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg 1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat 1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata 2041 agcttgatat cgaattcctg cagccccgac ggccagtgaa ttatcaacta tgtataataa 2101 agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa 2161 aaaacagact acataatact gtaaaacaca acatatccag tcactatggc ggccgtaagt 2221 tggcagcatc acccgacgca ctttgcgccg aataaatacc tgtgacggaa gatcacttcg 2281 cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc 2341 gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat 2401 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg 2461 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt 2521 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta 2581 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca 2641 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc 2701 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt 2761 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc 2821 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata 2881 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca 2941 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca 3001 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg 3061 gcagaatgct taatgaatta caacagtact gcgatgagtg gcaggggggg cgtaaacgcc 3121 gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg 3181 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg 3241 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg 3301 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg 3361 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa 3421 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc 3481 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg 3541 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc 3601 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat 3661 atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa 3721 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt 3781 atacacagcc agtctgcagg tcgatacagt agaaattaca gaaactttat cacgtttagt 3841 aagtatagag gctgaaaatc cagatgaagc cgaacgactt gtaagagaaa agtataagag 3901 ttgtgaaatt gttcttgatg cagatgattt tcaggactat gacactagcg tatatgaata 3961 ggtagatgtt tttattttgt cacacaaaaa agaggctcgc acctcttttt cttatttctt 4021 tttatgattt aatacggcat tgaggacaat agcgagtagg ctggatacga cgattccgtt 4081 tgagaagaac atttggaagg ctgtcggtcg accatagtga ctggatatgt tgtgttttac 4141 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat 4201 tttacgtttc tcgttcaact tttctataca aagttgatag cttggcgtaa tcgggggatc 4261 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag 4321 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 4381 cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct 4441 aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 4501 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 4561 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4621 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 4681 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 4741 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 4801 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 4861 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4921 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4981 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 5041 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 5101 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 5161 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 5221 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 5281 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5341 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5401 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5461 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5521 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5581 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5641 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5701 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5761 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5821 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5881 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5941 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 6001 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 6061 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 6121 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 6181 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6241 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6301 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6361 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6421 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6481 ttccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt 6541 aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa 6601 aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac 6661 cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg 6721 tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta 6781 tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg 6841 aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt 6901 atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc 6961 aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc 7021 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt 7081 cgtc //