LOCUS       sequence-126144-        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible NanoLuc expression, pNBU2 backbone, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cels.2015.06"; journalName: "Cell Syst"; date: 
            "2015-07-29- 29"; volume: "1"; issue: "1"; pages: "62-71"
COMMENT     Sequence Label: pMM597
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(5235..5303,5304..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
//
