LOCUS Exported 7061 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION IPTG-inducible NanoLuc expression, pNBU2 backbone, AmpR. ACCESSION . VERSION . KEYWORDS pMM597 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7061) AUTHORS Mimee M, Tucker AC, Voigt CA, Lu TK TITLE Programming a Human Commensal Bacterium, Bacteroides thetaiotaomicron, to Sense and Respond to Stimuli in the Murine Gut Microbiota. JOURNAL Cell Syst. 2015 Jul 29;1(1):62-71. doi: 10.1016/j.cels.2015.06.001. PUBMED 26918244 REFERENCE 2 (bases 1 to 7061) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7061 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 990..2072 /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(1009..1028) /label=LacI-R /note="LacI, reverse primer" CDS 5235..6095 /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 complement(5453..5472) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" oriT 6226..6335 /note="incP origin of transfer" rep_origin 6506..6894 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" protein_bind 6968..6984 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." protein_bind 7001..7017 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." ORIGIN 1 atggttttta ctctggaaga ttttgttggc gattggcgtc agaccgcggg ttataatttg 61 gatcaagtcc tggaacaggg tggcgtaagc tctctgttcc agaacctggg tgtgagcgtg 121 acgccgattc agcgcatcgt tctgtccggc gagaacggtc tgaaaattga tattcatgtg 181 atcatcccgt acgaaggcct gagcggtgac caaatgggtc aaatcgagaa aatctttaaa 241 gtcgtctacc cagttgacga tcaccacttc aaggttatct tgcattacgg tacgctggtg 301 attgatggtg tgaccccgaa tatgattgac tatttcggcc gtccgtatga aggcattgcc 361 gtttttgacg gtaaaaagat caccgtcacc ggtaccctgt ggaatggcaa taagattatt 421 gacgagcgtc tgattaaccc ggacggcagc ctgctgttcc gcgtgaccat caacggtgtc 481 acgggttggc gtctgtgcga gcgcatcctg gcataatgaa ctgcacttgc tttgataatt 541 aatgataaac aatctaaaag cactctaatc gttatcggag tgcttttaga ttactaatca 601 aattgcttct actaattgcc tatcttccag tgatggaaca gcatttgtgc attggctgca 661 acaatcagcc ttgatctgga agaagcaatg aaagctgctg ttaagtctcc gaatcaggta 721 ttgttcctga caggtgtatt cccatccggt aaacgcggat actttgcagt tgatctgact 781 caggaataaa ttataaatta aggtaagaag attgtaggat aagctaatga aatagaaaaa 841 ggatgccgtc acacaacttg tcggcattct tttttgtttt attagttgaa aatatagtga 901 aaaagttgcc taaatatgta tgttaacaaa ttatttgtcg taactttgca ctccaaatct 961 gtttttaaca tatggcacta gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag 1021 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt 1081 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc 1141 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc 1201 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg 1261 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg 1321 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc 1381 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct 1441 ctgaccagac acccatcaac agtattattt tctcccatga ggacggtacg cgactgggcg 1501 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt 1561 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc 1621 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc 1681 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc 1741 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag 1801 tgggatacga cgataccgag gacagctcat gttatatccc gccgttaacc accatcaaac 1861 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc 1921 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg 1981 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 2041 gacaggtttc ccgactggaa agcgggcagt gagctttcct cggtaccaaa ttccagaaaa 2101 gaggcctccc gaaagggggg ccttttttcg ttttggtcct acttgtgcct gttctatttc 2161 cgaaccgacc gcttgtatga atccatcaaa attcgttttc tctatgttgg attccttgtt 2221 gctcatattg tgatgataat ttctacaaat atagtcattg gtaactatct atgaaactgt 2281 ttgatacttt tatagttgat taaacttgtt catggcattt gccttaatat catccgctat 2341 gtcaatgtag ggtttcatag ctttgtagtc gctgtgtccc gtccatttca tgaccacctg 2401 tgccgggatt ccgagagcca gcgcattgca gatgaatgtc cttcttcctg catgggtact 2461 gagcaaagcg tatttgggtg tgacttcatc aatacgttca tttcccttgt agtaggtttc 2521 ccgtacaggc tcgttgattt ctgccagttc gcccagctct ttcaggtaat cgttcatctt 2581 ctggttgctg atgacgggca gagccatgta attctcgaaa tggatgtcct tgtatttgtc 2641 cagtatggct ttgctgtatt tgttcagttc aatcgtcagg ctgtcggcag tcttgactgt 2701 ggttatttcg atgtggtcgg acttcacatc gcttcttttc agattgcgaa catccgaata 2761 ccgcaaactc gtaaagcagc agaacaggaa aacatcacgc acacgttcca ggtattgctt 2821 atccttgggt atctggtagt ctttcagctt gttcagttca tcccaagtca ggaagattac 2881 ttttttcgag gtggttttca gtttcggttt gaacgtatcg tatgcaatgt tctgatgatg 2941 tcctttcttg aagctccagc gcaggaacca tttgaggaat cccatttgct tgccgatggt 3001 gctgtttctc atatccttgg tgtcacgcag gaagttgacg tattcgttca atccaaactc 3061 gttgaaatag ttgaacgttg catcctcctt gaactctttg aggtggttcc tcactgctgc 3121 aaatttttca taggtggatg ccgtccagtt attctggtta ccgcactctt ttacaaactc 3181 atcgaacacc tcccaaaagc tgacaggggc ttcttccggc tgttcttcgc tggtgtcttt 3241 cattctcatg ttgaaagctt ccttcaactg ttgggtcgtt ggcatgacct cctgcacctc 3301 aaattccttg aaaatattct ggatttcggc atagtatttc agcaagtccg tattgatttc 3361 ggctgcactt tgctttagct tgttggtaca tccgctcttt acccgctgct tatctgcatc 3421 ccatttggct acgtcaatcc ggtagcccgt tgtaaactcg atgcgttggc tggcaaagat 3481 gacacgcata cggatgggta cgttctctac gattggcaca ccgttctttt tccggctctc 3541 caatgcaaaa atgatgttgc gcttgatatt cataattggg tgcgtttgaa attctacacc 3601 caaatataca cccaattatt gagatagcaa aagacattta gaaacattta cttttactct 3661 atattgtaat ttacacttga ttatcagtcg tttgcagtct tatgatattc tgtgaaagta 3721 taagttcgag agcctgtctc tccgcaaaaa acgctgaaaa tcagcagatt gcaaaacaaa 3781 caccctgttt tacacccaag aatgtaaagt cggctgtttt tgttttattt aagataatac 3841 aaccactaca taataaaaga gtagcgatat taaaagaatc cgatgagaaa agactaatat 3901 ttatctatcc attcagtttg atttttcagg actttacatc gtcctgaaag tatttgttgg 3961 taccggtacc gaggacgcgt aaacatttac agttgcatgt ggcctattgt ttttagccgt 4021 taaatatttt ataactatta aatagcgata caaattgttc gaaactaata ttgtttatat 4081 catatattct cgcatgtttt aaagctttat taaattgatt ttttgtaaac agtttttcgt 4141 actctttgtt aacccatttc attacaaaag tttcatattt ttttctctct ttaaatgcca 4201 tttttgctgg ctttcttttt aatacaatta atgtgctatc cactttaggt tttggatgga 4261 aataatacct aggaattttt gctaatatag aaatatctac ctctgccatt aacagcaatg 4321 ctagtgatct gtttgtatct aataacattt tagcaaaacc atattccact attaaataac 4381 ttattgtggc tgaactttca aaaacaattt ttcgaattat atttgtgctt atgttgtaag 4441 gtatgctgcc aaatatttta tatggattgt ggctaggaaa tgtaaatttc agtatatcat 4501 catttactat ttgatagtta ggataattta agagcttatt acgagttacc tcacataatt 4561 tagaatcaat ttctatcgcc gttacaaaat tacatctctt taccaatcca gcagtaaaat 4621 gacctttccc tgcacctatt tcaaagatgt tatctttttc atctaaactt atgcaattca 4681 ttattttttc tatgtgatat tttgaagtaa taaaattttg actatctttt atatttactt 4741 tgttcattat aacctctcct taatttattg catctctttt cgaatattta tgttttttga 4801 gaaaagaacg tactcatggt tcatcccgat atgcgtatcg gtctgtatat cagcaacttt 4861 ctatgtgttt caactacaat agtcatctat tctcatcttt ctgagtccac cccctgcaaa 4921 gcccctcttt acgacataaa aattcggtcg gaaaaggtat gcaaaagatg tttctctctt 4981 taagagaaac tcttcgggat gcaaaaatat gaaaataact ccaattcacc aaattatata 5041 gcgacttttt tacaaaatgc taaaatttgt tgatttccgt caagcaattg ttgagcaaaa 5101 atgtctttta cgataaaatg atacctcaat atcaactgtt tagcaaaacg atatttctct 5161 taaagagaga aacacctttt tgttcaccaa tccccgactt ttaatcccgc ggccatgatt 5221 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 5281 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 5341 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5401 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5461 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5521 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5581 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5641 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5701 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5761 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5821 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5881 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 5941 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 6001 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 6061 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcataac 6121 gcgtcaattc gagggggatc aattccgtga taggtgggct gcccttcctg gttggcttgg 6181 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca 6241 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc 6301 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag 6361 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata 6421 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaac ggaattgatc 6481 cggccacgat gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta 6541 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat 6601 gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc 6661 tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa 6721 cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta 6781 aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc 6841 cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatgggaa 6901 ttcccctcca ccgcggtggt acaaagaaaa ttcgacaaac tgttattttt ctatctattt 6961 atttgaattg tgagcggata acaattacct ttgtcggcaa ttgtgagcgg ataacaatta 7021 aataaagata ttctcgtcaa acaaatataa ataatataaa c //