LOCUS Exported 3838 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expression of A2NT sgRNA from the arabinose-inducible promoter. ACCESSION . VERSION . KEYWORDS pAN-PBAD-sgRNA-A2NT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3838) AUTHORS Nielsen AA, Voigt CA TITLE Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. JOURNAL Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735. PUBMED 25422271 REFERENCE 2 (bases 1 to 3838) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3838 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 91..112 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" CDS complement(113..991) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1018..1302 /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 1225..1244 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" misc_RNA 1324..1399 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS 1421..1450 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1466..1483 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1539..1556) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1709..1755 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(1808..1825) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1979..2567) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2059..2078) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2738..3598) /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 3361..3380 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3599..3703) /gene="bla" /label=AmpR promoter primer_bind 3771..3789 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 3793..3836 /label=bacterial terminator /note="putative bacterial transcription terminator" ORIGIN 1 ttcagccaaa aaacttaaga ccgccggtct tgtccactac cttgcagtaa tgcggtggac 61 aggatcggcg gttttctttt ctcttctcaa gaattcgcgg ccgcttctag agttatgaca 121 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg 181 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc 241 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac 301 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca 361 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg 421 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga 481 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca 541 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc 601 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg 661 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc 721 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa 781 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa 841 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat 901 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg 961 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag 1021 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct 1081 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag 1141 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt 1201 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc 1261 ctacctgacg ctttttatcg caactctcta ctgtttctcc ataataatac ccgtgtgacc 1321 cgtgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa 1381 agtggcaccg agtcggtgct tttttttgaa gcttgggccc gaacaaaaac tcatctcaga 1441 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc 1501 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa 1561 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct 1621 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc 1681 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg 1741 ggcctttcgt tttatctgtt gtttgtcggt gaactggatc cttactcgag tctagactgc 1801 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 1861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 1921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 1981 tccacaggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 2041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 2101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 2161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 2221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 2281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 2341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 2401 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 2461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 2521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 2581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 2641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 2701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 2761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 2821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 2881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 2941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 3001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 3061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 3121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 3181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 3241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 3301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 3361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 3421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 3481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 3541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 3601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 3661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 3721 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 3781 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctg //