LOCUS Exported 3684 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION U6 promoter driving Ebf.774 (F+E) sgRNA. ACCESSION . VERSION . KEYWORDS U6>Ebf.774(F+E) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3684) AUTHORS Stolfi A, Gandhi S, Salek F, Christiaen L TITLE Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9. JOURNAL Development. 2014 Nov;141(21):4115-20. doi: 10.1242/dev.114488. PUBMED 25336740 REFERENCE 2 (bases 1 to 3684) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3684 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 1431..1565 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1468..1487) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1522..1541 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1601..1620) /label=T7 /note="T7 promoter, forward primer" promoter complement(1602..1620) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1707..1724) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1878..2466) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1958..1977) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2637..3497) /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 3260..3279 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3498..3602) /gene="bla" /label=AmpR promoter primer_bind 3670..4 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 aggcccttac gtattaatta aggcgcgcct ggcgggtgta ttaaaccact aaacaaacaa 61 ttgccccaag ctctcttcac aattataaac actacaaatg tttggacaag agattagcgt 121 ggctgtgacg agtaatctca aaggcttggt gtaattgata ttttataaga agcagattaa 181 acttcaatac agttaacacc tcatttacaa aaaattggct gccaaaatcg ctaattaaca 241 catatttaaa acaatttcta cagatataca cagtatagta tgattactaa ctgcataata 301 aacaaacata tccaacagac actcactaat ctgccataac aagcttcaaa aacttaaact 361 cgaaatttta gtgaatcttt ttttttaatg aagattttat ttaaaaagtt aaaaatatta 421 cagttcaggt ataggtttac acctaatctt taataatccg aactaaattt taactattta 481 gaaacttttt caaccaaagt ttaaaaaaat agattcttcg cacgctaaaa ctatcattta 541 cacaaaaaaa tgcaacaaaa tgcagaaaaa aattacatta gagtttaggt tagttacctg 601 ctaatcatta taaactaact tccggcataa tattcatcta aaattagcaa taatcacgtt 661 ttacgctaaa atttgtgtaa aactaaactt cgtcctttgt caaggagaaa atttgactca 721 aaagctgcgc gcgcagggga gatccccaag cgagtgtttg ttacatcata atcatgtgga 781 aaaatcccct aataagtaaa aatacatatt ttttaatttt gggggcaaat aaaccgcttt 841 ttatgtctaa aaacgccaaa aatggatcgc gcgagcccaa aaacgcacaa ataacgtaca 901 gacagtgtct ctgcgtacac agacggtatt tcccctttaa attgagaact agacttaagc 961 acgcttataa gtctggaagg catccgatgg tatagatgct gagggttgga caacagggtt 1021 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg 1081 aaaaagtggc accgagtcgg tgcttttttt gaattccagc tgagcgccgg tcgctaccat 1141 taccagttgg tctggtgtca aaaataataa taaccgggca ggccatgtct gcccgtattt 1201 cgcgtaagga aatccattat gtactattta aaaaacacaa acttttggat gttcggttta 1261 ttctttttct tttacttttt tatcatggga gcctacttcc cgtttttccc gatttggcta 1321 catgacatca accatatcag caaaagtgat acgggtatta tttttgccgc tatttctctg 1381 ttctcgctat tattccaacc gctgtttggt ctgctttctg acaaactcgg aacttgttta 1441 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 1501 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 1561 ggatcgacaa agtcaaagcg gccatcagat ctgccggact ccctatagtg agtcgtatta 1621 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 1681 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 1741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 1801 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 1861 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 1921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 1981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 2041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 2101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 2161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 2221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 2281 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 2341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 2401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 2461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 2521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 2581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 2641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 2701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 2761 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag 2821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 2881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 2941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 3001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 3061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 3121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 3181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 3241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 3301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 3361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 3421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 3481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 3541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 3601 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta 3661 acctataaaa ataggcgtat cacg //