LOCUS Exported 3382 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cloning vector for expression of sgRNA-15xPBSw. ACCESSION . VERSION . KEYWORDS pAC1428-pX-sgRNA-15xPBSw SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3382) 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 3382) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3382 /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 645..785 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 645..774 /label=AAV2 ITR rep_origin 860..1315 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(947..966) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1157..1178 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1332..1351) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1451..1473 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1511..1529) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1597..1701 /gene="bla" /label=AmpR promoter CDS 1702..2562 /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(1920..1939) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2733..3321 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3222..3241 /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 agattgtata tagcctgtat atagcctgta tatagcctgt atatagcctg 481 tatataagat tgtatatagc ctgtatatag cctgtatata gcctgtatat agcctgtata 541 taagatcttt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtagc 601 gcgtgcgcca attctgcaga caaatggcgg cgcgccgcgg ccgcaggaac ccctagtgat 661 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt 721 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct 781 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat 841 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 901 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 961 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 1021 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt 1081 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 1141 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 1201 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 1261 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg 1321 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca 1381 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct 1441 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg 1501 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt 1561 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 1621 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 1681 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 1741 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 1801 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 1861 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 1921 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata 1981 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 2041 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 2101 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 2161 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 2221 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 2281 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 2341 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 2401 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc 2461 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 2521 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 2581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 2641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 2701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 2761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 2821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc 2881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 2941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 3001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 3061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 3121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 3181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 3241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 3301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 3361 tgctggcctt ttgctcacat gt //