LOCUS Exported 3270 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cloning vector for expression of sgRNA-5xPBSw. ACCESSION . VERSION . KEYWORDS pAC1379-pX-sgRNA-5xPBSw SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3270) AUTHORS Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H TITLE Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. JOURNAL Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. PUBMED 26768771 REFERENCE 2 (bases 1 to 3270) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3270 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 9..249 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 9..29 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 180..199 /label=LKO.1 5' /note="Human U6 promoter, forward primer" repeat_region 533..673 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 533..662 /label=AAV2 ITR rep_origin 748..1203 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(835..854) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1045..1066 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1220..1239) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1339..1361 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1399..1417) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1485..1589 /gene="bla" /label=AmpR promoter CDS 1590..2450 /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(1808..1827) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2621..3209 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3110..3129 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggcgcgccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg 61 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt 121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg 181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt 241 ggaaaggacg aaacaccggg tcttcgagaa gacctgttta agagctatgc tggaaacagc 301 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg 361 ccaattgggt ctccagattg tatatagcct gtatatagcc tgtatatagc ctgtatatag 421 cctgtatata agatcttttt ttgttttaga gctagaaata gcaagttaaa ataaggctag 481 tccgtagcgc gtgcgccaat tctgcagaca aatggcggcg cgccgcggcc gcaggaaccc 541 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga 601 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc 661 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 721 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg 781 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 841 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 901 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 961 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 1021 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 1081 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 1141 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa 1201 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 1261 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 1321 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 1381 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 1441 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 1501 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 1561 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 1621 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 1681 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 1741 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 1801 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 1861 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 1921 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 1981 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 2041 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 2101 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 2161 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 2221 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 2281 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg 2341 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 2401 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 2461 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 2521 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 2581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 2641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 2701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 2761 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 2821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 2881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 2941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 3001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 3061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 3121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 3181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 3241 tggccttttg ctggcctttt gctcacatgt //