LOCUS Exported 3362 bp ds-DNA circular SYN 08-SEP-2021 DEFINITION Backbone for expressing plant artificial miRNAs. ACCESSION . VERSION . KEYWORDS pRS300 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3362) AUTHORS Schwab R, Ossowski S, Riester M, Warthmann N, Weigel D TITLE Highly specific gene silencing by artificial microRNAs in Arabidopsis. JOURNAL Plant Cell. 2006 May . 18(5):1121-33. PUBMED 16531494 REFERENCE 2 (bases 1 to 3362) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 8, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3362 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..456 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(88..107) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 298..319 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 582..604 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 596..613 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 597..613 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 620..639 /label=T7 /note="T7 promoter, forward primer" promoter 620..638 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 663..679 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 664..680 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1118..1134) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1118..1134) /label=pBluescriptSK /note="For pBluescript vector" promoter complement(1171..1189) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1210..1226) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1210..1226) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1223..1245) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1234..1250 /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(1258..1288) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1303..1324 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1441..1458) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1612..2200) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1692..1711) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2371..3231) /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 2994..3013 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3232..3336) /gene="bla" /label=AmpR promoter ORIGIN 1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 421 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa 481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg 541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 601 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggta ccgggccccc 661 cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccc aaacacacgc 721 tcggacgcat attacacatg ttcatacact taatactcgc tgttttgaat tgatgtttta 781 ggaatatata tgtagagaga gcttccttga gtccattcac aggtcgtgat atgattcaat 841 tagcttccga ctcattcatc caaataccga gtcgccaaaa ttcaaactag actcgttaaa 901 tgaatgaatg atgcggtaga caaattggat cattgattct ctttgattgg actgaaggga 961 gctccctctc tcttttgtat tccaattttc ttgattaatc tttcctgcac aaaaacatgc 1021 ttgatccact aagtgacata tatgctgcct tcgtatatat agttctggta aaattaacat 1081 tttgggttta tctttattta aggcatcgcc atggggggat ccactagttc tagagcggcc 1141 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattt cgagcttggc 1201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 1261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 1321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 1381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 1441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 1501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 1561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 1621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 1681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 1741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 1801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 1861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 1921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 1981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 2041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 2101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 2161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 2221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 2281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 2341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 2401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 2461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 2521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 2581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 2641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 2701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 2761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 2821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 2881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 2941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 3001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 3061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 3121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 3181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 3241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 3301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 3361 tg //