LOCUS Exported 5130 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circuit: OR gate, the sgRNA used for the NOT gate was substituted with one designed to target the malT gene in the E. coli genome. ACCESSION . VERSION . KEYWORDS pAN-OR-MalT-3NT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5130) 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 5130) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5130 /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 1323..1398 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS 1419..1448 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(1534..1551) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1704..1750 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(1803..1820) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1974..2562) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2054..2073) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2733..3593) /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 3356..3375 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3594..3698) /gene="bla" /label=AmpR promoter terminator 4557..4628 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4644..4671 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_RNA 4768..4843 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_RNA 4983..5058 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" 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 atataatacc cgtgtgaccc 1321 gtgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa 1381 gtggcaccga gtcggtgctt tttttgaagc ttgggcccga acaaaaactc atctcagaag 1441 aggatctgaa tagcgccgtc gaccatcatc atcatcattg agtttaaacg gtctccagct 1501 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag 1561 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc 1621 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc 1681 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct 1741 ttcgttttat ctgttgtttg tcggtgaact ggatccttac tcgagtctag actgcaggct 1801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 1861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccac 1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 2401 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 2581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 2701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 2761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 2821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 2881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 2941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 3001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 3061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 3121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 3181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 3241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 3301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 3361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 3421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 3481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 3541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 3601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 3661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 3721 cctgacgtct aagaaaccat tattatcatg acattaacca gttgataagt ccctaacttt 3781 tacagctagc tcagtcctag gtattatgct agcctgaagt acgtctgagc gtgatacccg 3841 ctcactgaag atggcccggt agggccgaaa cgtacctcta caaataattt tgtttaacta 3901 tggactatgt ttgaaaggga gaaatactag atggcacgta ccccgagccg tagcagcatt 3961 ggtagcctgc gtagtccgca tacccataaa gcaattctga ccagcaccat tgaaatcctg 4021 aaagaatgtg gttatagcgg tctgagcatt gaaagcgttg cacgtcgtgc cggtgcaagc 4081 aaaccgacca tttatcgttg gtggaccaat aaagcagcac tgattgccga agtgtatgaa 4141 aatgaaagcg aacaggtgcg taaatttccg gatctgggta gctttaaagc cgatctggat 4201 tttctgctgc gtaatctgtg gaaagtttgg cgtgaaacca tttgtggtga agcatttcgt 4261 tgtgttattg cagaagcaca gctggaccct gcaaccctga cccagctgaa agatcagttt 4321 atggaacgtc gtcgtgagat gccgaaaaaa ctggttgaaa atgccattag caatggtgaa 4381 ctgccgaaag ataccaatcg tgaactgctg ctggatatga tttttggttt ttgttggtat 4441 cgcctgctga ccgaacagct gaccgttgaa caggatattg aagaatttac cttcctgctg 4501 attaatggtg tttgtccggg tacacagcgt taactagggc ccataccccc aggcatcaaa 4561 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga 4621 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac 4681 gcgacgtacg gtggaatctg attcgttacc aattgacatg atacgaaacg taccgtatcg 4741 ttaaggtata atacccgtgt gacccgtgtt ttagagctag aaatagcaag ttaaaataag 4801 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcggtacca 4861 aagacgaaca ataagacgct gaaaagcgtc ttttttcgtt ttggtccttt agaattccag 4921 aaatcatttt acaccaacgg gtcacacggg tattatgcta gccgccagct caatgaaaag 4981 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa 5041 gtggcaccga gtcggtgctt tttttctcgg taccaaattc cagaaaagag gcctcccgaa 5101 aggggggcct tttttcgttt tggtccgctt //