LOCUS Exported 4432 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION SthCascade R promoter targeting crRNA1 expression cassette for Zea mays. ACCESSION . VERSION . KEYWORDS PV564 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4432) AUTHORS Joshua K. Young, Stephen L. Gasior, Spencer Jones, Lijuan Wang, TITLE Repurposing of Type I-E CRISPR-Cascade for gene activation in plants JOURNAL Nature Communications Biology REFERENCE 2 (bases 1 to 4432) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4432 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1..17 /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 6..28 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 25..41 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 25..41 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 62..80 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1641..1657) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1642..1658) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(1682..1701) /label=T7 /note="T7 promoter, forward primer" promoter complement(1683..1701) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1708..1725) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1708..1724) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1717..1739) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 1866..2321 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1953..1972) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2163..2184 /label=F1ori-F /note="F1 origin, forward primer" promoter 2347..2451 /gene="bla" /label=AmpR promoter CDS 2452..3312 /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(2670..2689) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3483..4071 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3972..3991 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4225..4242 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4359..4380 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4395..4425 /label=lac promoter /note="promoter for the E. coli lac operon" ORIGIN 1 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg 61 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta 121 gaactagtca gaacaagtac ctcggcgtga tcatccagtg cctcgtgacc gagaccaaga 181 cctccgtgtc caggcacatc tacttcttct ccgctcgcgg caggatcgac cccctcgtgt 241 acctcgacga gttcctcagg aactcagagc ccgtgctcaa gagggtgaac aggaccggca 301 actcctcctc caacaagcag gagtaccagc tcctcaagga caacctcgtg aggtcctaca 361 acaaggccct caagaagaac gccccctact ccatcttcgc catcaagaac ggccccaagt 421 cccacatcgg taggcacctc atgacctcct tcctctcaat gaagggcctc accgagctca 481 ccaacgtggt gggcaactgg tccgacaaga gggccctgca ggaattcgcg gccgcaaaaa 541 aaaggatcac ccccgcgtgt gcgggaaaaa cgaccgagca agacggtgat ggaagtagac 601 cctcggatca cccccgcgtg tgcgggaaaa acaattcggt gcttgcggct cggtttataa 661 ggtacgctcc actcttgtga ttagtcaagg tgggacaaaa cgtttgcgta ttacccagcg 721 ctttgctcca cctacggact gctgtttggg cttcgtcgcg gtgcttcgta ttgggccgtg 781 ggccatgttt cttatggccc tctctgtgac gaagtttttg gcacatacgc cgctatgact 841 ttctctttta cttttttgca aatattctcc gtcaacgcgg cgctgacact gtgtcctact 901 gtacatagaa ggaaaaaagg tatataaaaa aataatttct aaacacgact tactgctagt 961 agaaattcga atagcatttt agcttttaca actgggcctg gaaattcttt ggacaccact 1021 attttacgac ttatcatcat cgtgtgttag agcacaagga actatacaag cgaggttgct 1081 tatttaagcg ggatcattat gtcaaggatt cgatggggct aacaaaaaca gctggaaaca 1141 cagccagatc tttgatgccc cctcgctcaa aacttgcatt ccttgtacat gcgctaacaa 1201 acagaagaaa tcccttttta caatacaatt ttactttcct attcaccctg ccatatatca 1261 tacaaagttg gccaggacat gcgcccatgg ccaccatctc taaccctaat ggtgccatca 1321 aactggcacc atgatgccgg ttatctcact accttgctgc aatactgaca ctctgacagc 1381 atgccaattt agctcagcga ttactggtat ccttcagggc ctgttcggtt gtaccggaat 1441 aagccaggat tcattccagc tcatcaaaat ctatataaat tagagaagta atccggctag 1501 gaattaatcc ggggctctaa tccggaacaa ccgaacaggg cctcaagcaa tcttgatttt 1561 ctggatcata gactgagggt gcatttttca gactttggca ttgattaatc taggttcatc 1621 attgtactct caaagcttat cgataccgtc gacctcgagg gggggcccgg tacccaattc 1681 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga 1741 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg 1801 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 1861 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 1921 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 1981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2161 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2221 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2281 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga 2341 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 2401 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 2461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 2521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 2581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 2641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 2701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 2761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 2821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 2881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 2941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 3001 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 3061 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 3121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 3181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 3241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 3301 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 3361 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 3421 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 3481 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 3541 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 3601 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 3661 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 3721 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 3781 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 3841 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 3901 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 3961 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 4021 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 4081 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 4141 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 4201 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca 4261 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 4321 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat 4381 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aa //