LOCUS       sequence-128008-        7080 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 7080)
  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 7080)
  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: pMM596
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    45..61
                     /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    101..117
                     /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)."
     CDS             1151..2233
                     /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(1170..1189)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             join(5396..5464,5465..6256)
                     /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(5614..5633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            6387..6496
                     /note="incP origin of transfer"
     rep_origin      6667..7055
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 tacaaagaaa attcgacaaa ctgttatttt tctatctatt taaattgtga gcggataaca
       61 atttgggtgg gaaactttag ttatgtacct ttgtcggcaa ttgtgagcgg ataacaatta
      121 aataaagata ttctcgtcaa acaaatataa ataatataaa catggttttt actctggaag
      181 attttgttgg cgattggcgt cagaccgcgg gttataattt ggatcaagtc ctggaacagg
      241 gtggcgtaag ctctctgttc cagaacctgg gtgtgagcgt gacgccgatt cagcgcatca
      301 ttctgtccgg cgagaacggt ctgaaaattg atattcatgt gatcatcccg tacgaaggcc
      361 tgagcggtga ccaaatgggt caaatcgaga aaatctttaa agtcgtctac ccagttgacg
      421 atcaccactt caaggttatc ttgcattacg gtacgctggt gattgatggt gtgaccccga
      481 atatgattga ctatttcggc cgtccgtatg aaggcattgc cgtttttgac ggtaaaaaga
      541 tcaccgtcac cggtaccctg tggaatggca ataagattat tgacgagcgt ctgattaacc
      601 cggacggcag cctgctgttc cgcgtgacca tcaacggtgt cacgggttgg cgtctgtgcg
      661 agcgcatcct ggcataatga actgcacttg ctttgataat taatgataaa caatctaaaa
      721 gcactctaat cgttatcgga gtgcttttag attactaatc aaattgcttc tactaattgc
      781 ctatcttcca gtgatggaac agcatttgtg cattggctgc aacaatcagc cttgatctgg
      841 aagaagcaat gaaagctgct gttaagtctc cgaatcaggt attgttcctg acaggtgtat
      901 tcccatccgg taaacgcgga tactttgcag ttgatctgac tcaggaataa attataaatt
      961 aaggtaagaa gattgtagga taagctaatg aaatagaaaa aggatgccgt cacacaactt
     1021 gtcggcattc ttttttgttt tattagttga aaatatagtg aaaaagttgc ctaaatatgt
     1081 atgttaacaa attatttgtc gtaactttgc actccaaatc tgtttttaac atatggcact
     1141 agtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     1201 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1261 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1321 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1381 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1441 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1501 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1561 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     1621 cagtattatt ttctcccatg aggacggtac gcgactgggc gtggagcatc tggtcgcatt
     1681 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     1741 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     1801 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     1861 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     1921 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1981 ggacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     2041 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     2101 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     2161 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2221 aagcgggcag tgagctttcc tcggtaccaa attccagaaa agaggcctcc cgaaaggggg
     2281 gccttttttc gttttggtcc tacttgtgcc tgttctattt ccgaaccgac cgcttgtatg
     2341 aatccatcaa aattcgtttt ctctatgttg gattccttgt tgctcatatt gtgatgataa
     2401 tttctacaaa tatagtcatt ggtaactatc tatgaaactg tttgatactt ttatagttga
     2461 ttaaacttgt tcatggcatt tgccttaata tcatccgcta tgtcaatgta gggtttcata
     2521 gctttgtagt cgctgtgtcc cgtccatttc atgaccacct gtgccgggat tccgagagcc
     2581 agcgcattgc agatgaatgt ccttcttcct gcatgggtac tgagcaaagc gtatttgggt
     2641 gtgacttcat caatacgttc atttcccttg tagtaggttt cccgtacagg ctcgttgatt
     2701 tctgccagtt cgcccagctc tttcaggtaa tcgttcatct tctggttgct gatgacgggc
     2761 agagccatgt aattctcgaa atggatgtcc ttgtatttgt ccagtatggc tttgctgtat
     2821 ttgttcagtt caatcgtcag gctgtcggca gtcttgactg tggttatttc gatgtggtcg
     2881 gacttcacat cgcttctttt cagattgcga acatccgaat accgcaaact cgtaaagcag
     2941 cagaacagga aaacatcacg cacacgttcc aggtattgct tatccttggg tatctggtag
     3001 tctttcagct tgttcagttc atcccaagtc aggaagatta cttttttcga ggtggttttc
     3061 agtttcggtt tgaacgtatc gtatgcaatg ttctgatgat gtcctttctt gaagctccag
     3121 cgcaggaacc atttgaggaa tcccatttgc ttgccgatgg tgctgtttct catatccttg
     3181 gtgtcacgca ggaagttgac gtattcgttc aatccaaact cgttgaaata gttgaacgtt
     3241 gcatcctcct tgaactcttt gaggtggttc ctcactgctg caaatttttc ataggtggat
     3301 gccgtccagt tattctggtt accgcactct tttacaaact catcgaacac ctcccaaaag
     3361 ctgacagggg cttcttccgg ctgttcttcg ctggtgtctt tcattctcat gttgaaagct
     3421 tccttcaact gttgggtcgt tggcatgacc tcctgcacct caaattcctt gaaaatattc
     3481 tggatttcgg catagtattt cagcaagtcc gtattgattt cggctgcact ttgctttagc
     3541 ttgttggtac atccgctctt tacccgctgc ttatctgcat cccatttggc tacgtcaatc
     3601 cggtagcccg ttgtaaactc gatgcgttgg ctggcaaaga tgacacgcat acggatgggt
     3661 acgttctcta cgattggcac accgttcttt ttccggctct ccaatgcaaa aatgatgttg
     3721 cgcttgatat tcataattgg gtgcgtttga aattctacac ccaaatatac acccaattat
     3781 tgagatagca aaagacattt agaaacattt acttttactc tatattgtaa tttacacttg
     3841 attatcagtc gtttgcagtc ttatgatatt ctgtgaaagt ataagttcga gagcctgtct
     3901 ctccgcaaaa aacgctgaaa atcagcagat tgcaaaacaa acaccctgtt ttacacccaa
     3961 gaatgtaaag tcggctgttt ttgttttatt taagataata caaccactac ataataaaag
     4021 agtagcgata ttaaaagaat ccgatgagaa aagactaata tttatctatc cattcagttt
     4081 gatttttcag gactttacat cgtcctgaaa gtatttgttg gtaccggtac cgaggacgcg
     4141 taaacattta cagttgcatg tggcctattg tttttagccg ttaaatattt tataactatt
     4201 aaatagcgat acaaattgtt cgaaactaat attgtttata tcatatattc tcgcatgttt
     4261 taaagcttta ttaaattgat tttttgtaaa cagtttttcg tactctttgt taacccattt
     4321 cattacaaaa gtttcatatt tttttctctc tttaaatgcc atttttgctg gctttctttt
     4381 taatacaatt aatgtgctat ccactttagg ttttggatgg aaataatacc taggaatttt
     4441 tgctaatata gaaatatcta cctctgccat taacagcaat gctagtgatc tgtttgtatc
     4501 taataacatt ttagcaaaac catattccac tattaaataa cttattgtgg ctgaactttc
     4561 aaaaacaatt tttcgaatta tatttgtgct tatgttgtaa ggtatgctgc caaatatttt
     4621 atatggattg tggctaggaa atgtaaattt cagtatatca tcatttacta tttgatagtt
     4681 aggataattt aagagcttat tacgagttac ctcacataat ttagaatcaa tttctatcgc
     4741 cgttacaaaa ttacatctct ttaccaatcc agcagtaaaa tgacctttcc ctgcacctat
     4801 ttcaaagatg ttatcttttt catctaaact tatgcaattc attatttttt ctatgtgata
     4861 ttttgaagta ataaaatttt gactatcttt tatatttact ttgttcatta taacctctcc
     4921 ttaatttatt gcatctcttt tcgaatattt atgttttttg agaaaagaac gtactcatgg
     4981 ttcatcccga tatgcgtatc ggtctgtata tcagcaactt tctatgtgtt tcaactacaa
     5041 tagtcatcta ttctcatctt tctgagtcca ccccctgcaa agcccctctt tacgacataa
     5101 aaattcggtc ggaaaaggta tgcaaaagat gtttctctct ttaagagaaa ctcttcggga
     5161 tgcaaaaata tgaaaataac tccaattcac caaattatat agcgactttt ttacaaaatg
     5221 ctaaaatttg ttgatttccg tcaagcaatt gttgagcaaa aatgtctttt acgataaaat
     5281 gatacctcaa tatcaactgt ttagcaaaac gatatttctc ttaaagagag aaacaccttt
     5341 ttgttcacca atccccgact tttaatcccg cggccatgat tgaaaaagga agagtatgag
     5401 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5461 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5521 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5581 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5641 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5701 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5761 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5821 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5881 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5941 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6001 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6061 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6121 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6181 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6241 gattaagcat tggtaactgt cagaccaagt ttactcataa cgcgtcaatt cgagggggat
     6301 caattccgtg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
     6361 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
     6421 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
     6481 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
     6541 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
     6601 gaccccgaag cagggttatg cagcggaaaa cggaattgat ccggccacga tgcgtccggc
     6661 gtagaggatc tgaagatcag cagttcaacc tgttgatagt acgtactaag ctctcatgtt
     6721 tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga actaaaccct
     6781 catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt tcacgtacta
     6841 agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt
     6901 tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg
     6961 gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca
     7021 attttgagtg acacaggaac acttaacggc tgacatggga attcccctcc accgcggtgg
//
