LOCUS Exported 3326 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cloning vector for expression of sgRNA-10xPBSb. ACCESSION . VERSION . KEYWORDS pAC1424-pX-sgRNA-10xPBSb SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3326) 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 3326) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3326 /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 589..729 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 589..718 /label=AAV2 ITR rep_origin 804..1259 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(891..910) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1101..1122 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1276..1295) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1395..1417 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1455..1473) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1541..1645 /gene="bla" /label=AmpR promoter CDS 1646..2506 /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(1864..1883) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2677..3265 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3166..3185 /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 ctccagattt gatatagcct tgatatagcc ttgatatagc cttgatatag 421 ccttgatata agatttgata tagccttgat atagccttga tatagccttg atatagcctt 481 gatataagat ctttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg 541 tagcgcgtgc gccaattctg cagacaaatg gcggcgcgcc gcggccgcag gaacccctag 601 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 661 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct 721 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 781 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt 1021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 1081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 1141 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 1201 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt 1261 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 1321 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 1381 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 1441 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 1501 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 1561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 1621 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 1681 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 1741 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 1801 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 1861 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 1921 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 1981 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 2041 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 2101 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 2161 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 2221 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 2281 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 2341 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa 2401 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 2461 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 2521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 2581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 2641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 2701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 2761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 2821 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 2881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 2941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 3001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 3061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 3121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 3181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 3241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 3301 cttttgctgg ccttttgctc acatgt //