LOCUS Exported 3906 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION sgRNA entry vector. ACCESSION . VERSION . KEYWORDS pWS082 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3906) TITLE Ellis Lab Yeast CRISPR Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 3906) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3906 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 116..135 /label=pBR322ori-F /note="pBR322 origin, forward primer" RBS 985..996 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1003..1716 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 1728..1799 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1815..1842 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_RNA 1856..1931 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS complement(2465..3325) /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 3088..3107 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3326..3430) /gene="bla" /label=AmpR promoter rep_origin 3533..215 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 301 ctgtggataa ccgtgcggcc gcccctgata tcaacaacag aggacatatg ccctacctcc 361 atgatctact gacgtccctg aggctgcaat tcatgtaatg gggcagtatc cgcggcaagt 421 cctagtgcaa tggcggtttt ttaccctcgt tctgaagaag aggcgacgcg ggtgcggtca 481 tcactaatgt ggaaattggt aagactctcg ggcctccgcc tttaggcggt gcttactctt 541 tcataaaggg gctgttagtt atgcaacgat gtgcttcagt attacatttt ttgccttcaa 601 cgccttgatt gttctatttt tgctaataat aaatctattt catcggacta aaagtccatt 661 agttgtaagc ggatttagct cagttgggag agcgccagac tgaagaaaaa cttcggtcaa 721 gtcatctgga ggtcctgtgt tcgatccaca gaattcgcag atggccggca tggtcccagc 781 ctcctcgctg gcgccggctg ggcaacacct tcgggtggcg aatgggactt gagacggaaa 841 gtgaaacgtg atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg 901 cggcaattca catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc 961 aggttcggat cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag 1021 ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag 1081 ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc 1141 atctgtacta ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat 1201 ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc 1261 gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac 1321 aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa 1381 ggcattgact ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac 1441 agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa 1501 attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact 1561 ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt 1621 ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca 1681 gcgggcatca cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa 1741 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct 1801 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacg tctcagtttt 1861 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac 1921 cgagtcggtg ctttttttat tttttgtcac tattgttatg taaaatgcca cctctgacag 1981 tatggaacgc aaacttctgt ctagtggata gctggacatt ccaggcagtg cctctgccgc 2041 cggacccctc tcgtgattgg gtagctggac atgcccttgt aagatataac aagagcctgc 2101 ctgtctaatg atctcacggc gaaagtcggg gagacagcag cggctgcaga cattataccg 2161 caacaacact aaggtgagat aactccgtac attgactacg cgttcctcta gaccttactt 2221 gaccggatac agtgtctttg acacgtttat gggttacagc aatcacatcc aagactggct 2281 atgcacgaag caactcttga gtgttaaaat gttgacccct gtatttggga tgcgggtagt 2341 agatgagtgc aggatatcta cagcggccgc gattatcaaa aaggatcttc acctagatcc 2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 2581 gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa 2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 2761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 3061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 3361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 3421 gggttccgcg cacatttccc cgaaaagtgc cacctgtcat gaccaaaatc ccttaacgtg 3481 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 3541 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 3601 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 3661 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 3721 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 3781 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 3841 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 3901 aactga //