LOCUS pYLsgRNA-OsU6a 3289 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-OsU6a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3289)
AUTHORS Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H,
Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X,
Dong Z, Liu YG
TITLE A robust CRISPR/Cas9 system for convenient, high-efficiency
multiplex genome editing in monocot and dicot plants.
JOURNAL Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi:
10.1016/j.molp.2015.04.007.
PUBMED 25917172
REFERENCE 2 (bases 1 to 3289)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..3289
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 20..95
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(643..660)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(643..659)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(652..674)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(868..887)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 987..1009
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1047..1065)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1133..1237
/gene="bla"
/label=AmpR promoter
CDS 1238..2098
/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(1456..1475)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2269..2857
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2758..2777
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3011..3028
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3145..3166
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3181..3211
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3219..3235
/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 3224..3246
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3243..3259
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3243..3259
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 ggatccagcg tgggtctcgg ttttagagct agaaatagca agttaaaata aggctagtcc
61 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcaagagct tggagtggat
121 ggaattttcc tccgttttac ctgtggaatc ggcagcaaag gattttttcc tgtagttttc
181 ccacaaccat tttttaccat ccgaatgata ggataggaaa aatatccaag tgaacagtat
241 tcctataaaa ttcccgtaaa aagcctgcaa tccgaatgag ccctgaagtc tgaactagcc
301 ggtcacctgt acaggctatc gagatgccat acaagagacg gtagtaggaa ctaggaagac
361 gatggttgat tcgtcaggcg aaatcgtcgt cctgcagtcg catctatggg cctggacgga
421 ataggggaaa aagttggccg gataggaggg aaaggcccag gtgcttacgt gcgaggtagg
481 cctgggctct cagcacttcg attcgttggc accggggtag gatgcaatag agagcaacgt
541 ttagtaccac ctcgcttagc tagagcaaac tggactgcct tatatgcgcg ggtgctggct
601 tggctgccga gagacctctg aagataacat actaagcttg gcactggccg tcgttttaca
661 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
721 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
781 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
841 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
901 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
961 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
1021 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
1081 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
1141 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
1201 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
1261 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
1321 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
1381 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
1441 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
1501 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
1561 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
1621 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
1681 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
1741 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
1801 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
1861 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
1921 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
1981 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
2041 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
2101 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
2161 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
2221 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
2281 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
2341 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
2401 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
2461 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
2521 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
2581 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
2641 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
2701 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
2761 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
2821 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
2881 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
2941 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
3001 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
3061 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
3121 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
3181 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
3241 cacaggaaac agctatgacc atgattacga attcgagctc ggtacccgg
//
