LOCUS Exported 4835 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION bacterial expression of AcrIIA1. ACCESSION . VERSION . KEYWORDS pRAU168 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4835) AUTHORS Rauch BJ, Silvis MR, Hultquist JF, Waters CS, McGregor MJ, Krogan NJ, Bondy-Denomy J TITLE Inhibition of CRISPR-Cas9 with Bacteriophage Proteins. JOURNAL Cell. 2016 Dec 22. pii: S0092-8674(16)31683-X. doi: 10.1016/j.cell.2016.12.009. PUBMED 28041849 REFERENCE 2 (bases 1 to 4835) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4835 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 17..301 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 224..243 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" terminator 879..965 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1057..1084 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1103..1194 /gene="bla" /label=AmpR promoter CDS 1195..2055 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1413..1432) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2097..2552 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2184..2203) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2394..2415 /label=F1ori-F /note="F1 origin, forward primer" rep_origin 2663..3251 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3152..3171 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3405..3422 /label=L4440 /note="L4440 vector, forward primer" primer_bind 3588..3610 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3748..3767 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(3947..4825) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc 61 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg 121 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc 181 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta 241 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca 301 tacccgtttt tttgggctag caggaggaat tcaccatgac tattaaatta ttagatgaat 361 tcttaaaaaa gcatgatttg acgaggtatc agttaagcaa attaactggt atctcacaaa 421 acactttaaa agaccagaac gaaaaaccgt taaataaata tactgtttca atactacgct 481 cgctatcgtt gatttcgggt ttatctgtat cagatgtttt gtttgaatta gaagacatag 541 aaaaaaattc tgacgatctt gcaggattta aacacctgtt agacaagtac aaactctcat 601 ttcctgcaca agaattcgaa ttatactgct taatcaaaga gtttgagtct gctaatattg 661 aagtacttcc ttttacgttc aatagatttg aaaacgaaga acatgtaaat ataaaaaaag 721 atgtttgtaa agcgttagaa aatgctatca ccgtgttaaa agagaagaaa aatgagttac 781 tttgataagt ttaaaaggaa ggaactcaga agtgaaacgc cgtagcgccg atggtagtgt 841 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt 901 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga 961 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag 1021 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc 1081 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg 1141 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 1201 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 1261 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 1321 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 1381 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt 1441 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 1501 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 1561 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 1621 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 1681 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 1741 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 1801 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 1861 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 1921 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 1981 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 2041 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc 2101 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2161 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2221 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 2281 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc 2341 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2401 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg 2461 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2521 gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg 2581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 2641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 2821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 2941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 3001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 3061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 3121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 3181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 3241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 3301 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 3361 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 3421 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 3481 accgcatagg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg 3541 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 3601 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa 3661 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 3721 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 3781 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt 3841 gacagcttat catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa 3901 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga 3961 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg 4021 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga 4081 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt 4141 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg 4201 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt 4261 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact 4321 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct 4381 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat 4441 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt 4501 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc 4561 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac 4621 ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga 4681 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt 4741 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca 4801 ggggatcatt ttgcgcttca gccatacttt tcata //