LOCUS Exported 2218 bp ds-DNA linear SYN 21-MAY-2021 DEFINITION empty sgRNA cloning vector with MS2-humanA3a. ACCESSION . VERSION . KEYWORDS pLL4 sgRNA(MS2)-MS2-hA3a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2218) AUTHORS Liu LD, Huang M, Dai P, Liu T, Fan S, Cheng X, Zhao Y, Yeap LS, Meng FL TITLE Intrinsic Nucleotide Preference of Diversifying Base Editors Guides Antibody Ex Vivo Affinity Maturation. JOURNAL Cell Rep. 2018 Oct 23;25(4):884-892.e3. doi: 10.1016/j.celrep.2018.09.090. PUBMED 30355495 REFERENCE 2 (bases 1 to 2218) AUTHORS . TITLE Direct Submission JOURNAL Exported May 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2218 /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" promoter 341..445 /gene="bla" /label=AmpR promoter primer_bind complement(664..683) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1291..1879 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1780..1799 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2033..2050 /label=L4440 /note="L4440 vector, forward primer" misc_RNA 2093..2111 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" misc_RNA 2163..2181 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" 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 ggtcttctcg tgatacgcct atttttatag gttaatgtca tgataataat 301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcccaa 901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagtaatttg 961 gagccggtga gcgtggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 1021 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 1081 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 1141 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 1201 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 1261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 1321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 1381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 1441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 1501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 1561 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 1621 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 1681 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 1741 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 1801 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 1861 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 1921 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 1981 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 2041 agtgagcgag gaagcggaag agcgcgagaa gacctgtttt agagctaggc caacatgagg 2101 atcacccatg tctgcagggc ctagcaagtt aaaataaggc tagtccgtta tcaacttggc 2161 caacatgagg atcacccatg tctgcagggc caagtggcac cgagtcggtg cttttttt //