LOCUS Exported 2279 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gRNA that guides Cas9 and APOBEC complexes to L138 in eGFP.. ACCESSION . VERSION . KEYWORDS eGFP L138 gRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2279) AUTHORS Martin AS, Salamango DJ, Serebrenik AA, Shaban NM, Brown WL, Harris RS TITLE A panel of eGFP reporters for single base editing by APOBEC-Cas9 editosome complexes. JOURNAL Sci Rep. 2019 Jan 24;9(1):497. doi: 10.1038/s41598-018-36739-9. PUBMED 30679582 REFERENCE 2 (bases 1 to 2279) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2279 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 82..322 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 82..102 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 253..272 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 351..426 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(530..1118) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(610..629) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1289..2149) /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 1912..1931 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2150..2254) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc 61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata 121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta 181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta 241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct 301 ttatatatct tgtggaaagg acgaaacacc caacatttca gggcacaagc gttttagagc 361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt 421 cggtgctttt tttaagcttg ggccgctcga ggtacctctc tacatatgac atgtgagcaa 481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 1861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 1921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct //