LOCUS Exported 3075 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Empty vector for cloning LbCpf1-spCas9 MS2-fgRNA. ACCESSION . VERSION . KEYWORDS pFGRNA-LbCpf1-spCas9-MS2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3075) AUTHORS Kweon J, Jang AH, Kim DE, Yang JW, Yoon M, Rim Shin H, Kim JS, Kim Y TITLE Publisher Correction: Fusion guide RNAs for orthogonal gene manipulation with Cas9 and Cpf1. JOURNAL Nat Commun. 2018 Jan 16;9(1):303. doi: 10.1038/s41467-017-02686-8. PUBMED 29335559 REFERENCE 2 (bases 1 to 3075) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3075 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" primer_bind 344..362 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(400..422) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 522..541 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 735..757 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 749..766 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 750..766 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_RNA 812..830 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" misc_RNA 882..900 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" primer_bind complement(1066..1083) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1237..1825) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1317..1336) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1996..2856) /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 2619..2638 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2857..2961) /gene="bla" /label=AmpR promoter ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg aatttctact aagtgtagat tgagaccaca tttccccgaa aagtgccacc 301 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag 361 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 421 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 481 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt 541 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg 601 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc 661 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt 721 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttgcatg 781 cctgcagggt ctctgtttta gagctaggcc aacatgagga tcacccatgt ctgcagggcc 841 tagcaagtta aaataaggct agtccgttat caacttggcc aacatgagga tcacccatgt 901 ctgcagggcc aagtggcacc gagtcggtgc ttttttttct agattcgcga tgtacgggcc 961 agatatacgc gttgacattg attattgact agttgtcttc ctgcattaat gaatcggcca 1021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 1081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 1141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 1201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 1261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 1321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 1381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 1441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 1501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 1561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 1621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 1681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 1741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 1801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 1861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 1921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 1981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 2041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 2101 cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga 2161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 2221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 2281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 2341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 2401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 2461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 2521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 2581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 2641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 2701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 2761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 2821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 2881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 2941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacgaaga 3001 ctcaattgtc gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg 3061 gccgccccct tcacc //