LOCUS       Exported                7059 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  IPTG-inducible NanoLuc expression, pNBU2 backbone, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    pMM585
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  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 7059)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7059
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    68..84
                     /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             1130..2212
                     /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(1149..1168)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5375..6235
                     /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(5593..5612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            6366..6475
                     /note="incP origin of transfer"
     rep_origin      6646..7034
                     /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 atttgaattg tgagcggata acaattacct ttgtcggcaa ataaagatat tctcgtcaaa
      121 caaatataaa taatataaac atggttttta ctctggaaga ttttgttggc gattggcgtc
      181 agaccgcggg ttataatttg gatcaagtcc tggaacaggg tggcgtaagc tctctgttcc
      241 agaacctggg tgtgagcgtg acgccgattc agcgcatcgt tctgtccggc gagaacggtc
      301 tgaaaattga tattcatgtg atcatcccgt acgaaggcct gagcggtgac caaatgggtc
      361 aaatcgagaa aatctttaaa gtcgtctacc cagttgacga tcaccacttc aaggttatct
      421 tgcattacgg tacgctggtg attgatggtg tgaccccgaa tatgattgac tatttcggcc
      481 gtccgtatga aggcattgcc gtttttgacg gtaaaaagat caccgtcacc ggtaccctgt
      541 ggaatggcaa taagattatt gacgagcgtc tgattaaccc ggacggcagc ctgctgttcc
      601 gcgtgaccat caacggtgtc acgggttggc gtctgtgcga gcgcatcctg gcataatgaa
      661 ctgcacttgc tttgataatt aatgataaac aatctaaaag cactctaatc gttatcggag
      721 tgcttttaga ttactaatca aattgcttct actaattgcc tatcttccag tgatggaaca
      781 gcatttgtgc attggctgca acaatcagcc ttgatctgga agaagcaatg aaagctgctg
      841 ttaagtctcc gaatcaggta ttgttcctga caggtgtatt cccatccggt aaacgcggat
      901 actttgcagt tgatctgact caggaataaa ttataaatta aggtaagaag attgtaggat
      961 aagctaatga aatagaaaaa ggatgccgtc acacaacttg tcggcattct tttttgtttt
     1021 attagttgaa aatatagtga aaaagttgcc taaatatgta tgttaacaaa ttatttgtcg
     1081 taactttgca ctccaaatct gtttttaaca tatggcacta gtggtgaatg tgaaaccagt
     1141 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     1201 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     1261 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     1321 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     1381 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     1441 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     1501 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     1561 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     1621 ggacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     1681 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     1741 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     1801 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     1861 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     1921 tggtgcggat atctcggtag tgggatacga cgataccgag gacagctcat gttatatccc
     1981 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     2041 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     2101 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2161 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagctttcct
     2221 cggtaccaaa ttccagaaaa gaggcctccc gaaagggggg ccttttttcg ttttggtcct
     2281 acttgtgcct gttctatttc cgaaccgacc gcttgtatga atccatcaaa attcgttttc
     2341 tctatgttgg attccttgtt gctcatattg tgatgataat ttctacaaat atagtcattg
     2401 gtaactatct atgaaactgt ttgatacttt tatagttgat taaacttgtt catggcattt
     2461 gccttaatat catccgctat gtcaatgtag ggtttcatag ctttgtagtc gctgtgtccc
     2521 gtccatttca tgaccacctg tgccgggatt ccgagagcca gcgcattgca gatgaatgtc
     2581 cttcttcctg catgggtact gagcaaagcg tatttgggtg tgacttcatc aatacgttca
     2641 tttcccttgt agtaggtttc ccgtacaggc tcgttgattt ctgccagttc gcccagctct
     2701 ttcaggtaat cgttcatctt ctggttgctg atgacgggca gagccatgta attctcgaaa
     2761 tggatgtcct tgtatttgtc cagtatggct ttgctgtatt tgttcagttc aatcgtcagg
     2821 ctgtcggcag tcttgactgt ggttatttcg atgtggtcgg acttcacatc gcttcttttc
     2881 agattgcgaa catccgaata ccgcaaactc gtaaagcagc agaacaggaa aacatcacgc
     2941 acacgttcca ggtattgctt atccttgggt atctggtagt ctttcagctt gttcagttca
     3001 tcccaagtca ggaagattac ttttttcgag gtggttttca gtttcggttt gaacgtatcg
     3061 tatgcaatgt tctgatgatg tcctttcttg aagctccagc gcaggaacca tttgaggaat
     3121 cccatttgct tgccgatggt gctgtttctc atatccttgg tgtcacgcag gaagttgacg
     3181 tattcgttca atccaaactc gttgaaatag ttgaacgttg catcctcctt gaactctttg
     3241 aggtggttcc tcactgctgc aaatttttca taggtggatg ccgtccagtt attctggtta
     3301 ccgcactctt ttacaaactc atcgaacacc tcccaaaagc tgacaggggc ttcttccggc
     3361 tgttcttcgc tggtgtcttt cattctcatg ttgaaagctt ccttcaactg ttgggtcgtt
     3421 ggcatgacct cctgcacctc aaattccttg aaaatattct ggatttcggc atagtatttc
     3481 agcaagtccg tattgatttc ggctgcactt tgctttagct tgttggtaca tccgctcttt
     3541 acccgctgct tatctgcatc ccatttggct acgtcaatcc ggtagcccgt tgtaaactcg
     3601 atgcgttggc tggcaaagat gacacgcata cggatgggta cgttctctac gattggcaca
     3661 ccgttctttt tccggctctc caatgcaaaa atgatgttgc gcttgatatt cataattggg
     3721 tgcgtttgaa attctacacc caaatataca cccaattatt gagatagcaa aagacattta
     3781 gaaacattta cttttactct atattgtaat ttacacttga ttatcagtcg tttgcagtct
     3841 tatgatattc tgtgaaagta taagttcgag agcctgtctc tccgcaaaaa acgctgaaaa
     3901 tcagcagatt gcaaaacaaa caccctgttt tacacccaag aatgtaaagt cggctgtttt
     3961 tgttttattt aagataatac aaccactaca taataaaaga gtagcgatat taaaagaatc
     4021 cgatgagaaa agactaatat ttatctatcc attcagtttg atttttcagg actttacatc
     4081 gtcctgaaag tatttgttgg taccggtacc gaggacgcgt aaacatttac agttgcatgt
     4141 ggcctattgt ttttagccgt taaatatttt ataactatta aatagcgata caaattgttc
     4201 gaaactaata ttgtttatat catatattct cgcatgtttt aaagctttat taaattgatt
     4261 ttttgtaaac agtttttcgt actctttgtt aacccatttc attacaaaag tttcatattt
     4321 ttttctctct ttaaatgcca tttttgctgg ctttcttttt aatacaatta atgtgctatc
     4381 cactttaggt tttggatgga aataatacct aggaattttt gctaatatag aaatatctac
     4441 ctctgccatt aacagcaatg ctagtgatct gtttgtatct aataacattt tagcaaaacc
     4501 atattccact attaaataac ttattgtggc tgaactttca aaaacaattt ttcgaattat
     4561 atttgtgctt atgttgtaag gtatgctgcc aaatatttta tatggattgt ggctaggaaa
     4621 tgtaaatttc agtatatcat catttactat ttgatagtta ggataattta agagcttatt
     4681 acgagttacc tcacataatt tagaatcaat ttctatcgcc gttacaaaat tacatctctt
     4741 taccaatcca gcagtaaaat gacctttccc tgcacctatt tcaaagatgt tatctttttc
     4801 atctaaactt atgcaattca ttattttttc tatgtgatat tttgaagtaa taaaattttg
     4861 actatctttt atatttactt tgttcattat aacctctcct taatttattg catctctttt
     4921 cgaatattta tgttttttga gaaaagaacg tactcatggt tcatcccgat atgcgtatcg
     4981 gtctgtatat cagcaacttt ctatgtgttt caactacaat agtcatctat tctcatcttt
     5041 ctgagtccac cccctgcaaa gcccctcttt acgacataaa aattcggtcg gaaaaggtat
     5101 gcaaaagatg tttctctctt taagagaaac tcttcgggat gcaaaaatat gaaaataact
     5161 ccaattcacc aaattatata gcgacttttt tacaaaatgc taaaatttgt tgatttccgt
     5221 caagcaattg ttgagcaaaa atgtctttta cgataaaatg atacctcaat atcaactgtt
     5281 tagcaaaacg atatttctct taaagagaga aacacctttt tgttcaccaa tccccgactt
     5341 ttaatcccgc ggccatgatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5401 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5461 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5521 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5581 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5641 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5701 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5761 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5821 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5881 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5941 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6001 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6061 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6121 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6181 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6241 agaccaagtt tactcataac gcgtcaattc gagggggatc aattccgtga taggtgggct
     6301 gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc
     6361 ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     6421 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     6481 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     6541 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     6601 agcggaaaac ggaattgatc cggccacgat gcgtccggcg tagaggatct gaagatcagc
     6661 agttcaacct gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt
     6721 ttaacgtact aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc
     6781 tcatggctaa cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata
     6841 aaattaatat aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa
     6901 aagaatatat aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg
     6961 atgtaacgca ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca
     7021 cttaacggct gacatgggaa ttcccctcca ccgcggtgg
//
