LOCUS Exported 2210 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo golden gate assembly DE part for gQi gRNA (guide RNA for S. pyogenes Cas9; sequence from DOI: 10.1016/j.cell.2013.02.022). Please see Supplemental Documents for annotated Genbank file.. ACCESSION . VERSION . KEYWORDS UC16m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2210) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 2210) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2210 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(10..598) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(90..109) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(769..1629) /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 1392..1411 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1630..1734) /gene="bla" /label=AmpR promoter primer_bind 1802..1820 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 1824..1867 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 1870..1891 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" misc_RNA 1930..2005 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 2023..2043 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(2049..2066) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gctggcgttt ttccacaggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 61 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 901 gataccgcgc gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 1021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 1081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 1141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 1201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 1261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 1321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 1381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 1441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 1501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 1561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 1621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 1681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 1741 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 1801 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga 1861 tgatttctgg aattcgcggc cgcttctaga gactagtggg tctcatacta actttcagtt 1921 tagcggtctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact 1981 tgaaaaagtg gcaccgagtc ggtgcttttt taggtagaga cctactagta gcggccgctg 2041 caggcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 2101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 2161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt //