LOCUS       sequence-201654-        5034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses taz from a LacI regulated promoter under inducible IPTG 
            control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5034)
  AUTHORS   Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ
  TITLE     Phosphatase activity tunes two-component system sensor detection 
            threshold.
  JOURNAL   Nat Commun. 2018 Apr 12;9(1):1433. doi: 10.1038/s41467-018-03929-y.
  PUBMED    29650958
REFERENCE   2  (bases 1 to 5034)
  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"; journalName: "Nat 
            Commun."; date: "2018-04-12"; volume: "9(1):1433. doi"; pages: " 
            10.1038/s41467-018-03929-y"
COMMENT     Sequence Label: pBL132
FEATURES             Location/Qualifiers
     source          1..5034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             149..940
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      952..1046
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1160..1705
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1606..1625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1822..1839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1878..1955
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1956..3038
                     /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(1975..1994)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3044..3130
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(3303..3308,3309..3324,3325..3331)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3339..3355
                     /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)."
     terminator      4869..4940
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4956..4983
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
       61 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
      121 tcaagagaca ggatgaggat cgtttcgtat gagggaagcg gtgatcgccg aagtatcgac
      181 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
      241 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
      301 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
      361 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
      421 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
      481 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
      541 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
      601 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
      661 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
      721 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
      781 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
      841 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
      901 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa gagctcgctt ggactcctgt
      961 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1021 gccgggcgtt ttttattggt gagaatccaa gcactagtaa caacttatat cgtatggggc
     1081 tgacttcagg tgctacattt gaagagataa attgcactga aatctagtaa tattttatct
     1141 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     1201 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     1261 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     1321 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     1381 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     1441 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     1501 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     1561 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     1621 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     1681 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     1741 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     1801 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcccgaag
     1861 cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
     1921 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     1981 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     2041 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     2101 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     2161 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     2221 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     2281 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     2341 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     2401 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     2461 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     2521 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     2581 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     2641 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     2701 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     2761 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     2821 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     2881 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     2941 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3001 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagg catcaaataa aacgaaaggc
     3061 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     3121 taggacaaat ccgccgccct agacctaggg cgttcggctg cggcgagcgg tatcagctca
     3181 ctcaaaggcg gtaatacgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     3241 tctggataat gttttttgcg ccgacatcat aacggttctg gctaatattc tgaaatgagc
     3301 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     3361 cacaagaaca cttagagaac aaacgggaga gtatctaaaa tgattaaccg tatccgcgta
     3421 gtcacgctgt tggtaatggt gctgggggta ttcgcactgt tacagcttat ttccggcagt
     3481 ctgttttttt cttcccttca ccatagccag aagagctttg tggtttccaa tcaattacgg
     3541 gaacagcagg gcgagctgac gtcaacctgg gatttaatgc tgcaaacgcg cattaacctg
     3601 agtcgttcag cggtacggat gatgatggat tcctccaatc aacaaagtaa cgccaaagtt
     3661 gaattgctcg atagcgccag gaaaacattg gcgcaggcag cgacgcatta taaaaaattc
     3721 aaaagcatgg caccgttacc tgaaatggtc gctaccagtc gtaatattga tgaaaaatat
     3781 aaaaactatt acacagcgtt aactgaactg attgattatc tagattatgg caatactgga
     3841 gcttatttcg ctcagccaac ccagggaatg caaaatgcaa tgggcgaagc gtttgctcag
     3901 tacgccctca gcagtgaaaa actgtatcgc gatatcgtca ctgacaacgc agatgattac
     3961 cgatttgccc agtggcaact ggcggttatc gcgctggtgg tggtattgat tctgctggtg
     4021 gcgtggtacg gcattcgccg tatgttgctt actccgctgg caaaaattat tgctcacatt
     4081 cgcgaaatcg ccggtggtaa cctggcgaat accctgacca ttgacgggcg cagtgaaatg
     4141 ggcgacctgg cgcagagcgt ttcacatatg gcggctggtg ttaagcaact ggcggatgac
     4201 cgcacgctgc tgatggcggg ggtaagtcac gacttgcgca cgccgctgac gcgtattcgc
     4261 ctggcgactg agatgatgag cgagcaggat ggctatctgg cagaatcgat caataaagat
     4321 atcgaagagt gcaacgccat cattgagcag tttatcgact acctgcgcac cgggcaggag
     4381 atgccgatgg aaatggcgga tcttaatgca gtactcggtg aggtgattgc tgccgaaagt
     4441 ggctatgagc gggaaattga aaccgcgctt taccccggca gcattgaagt gaaaatgcac
     4501 ccgctgtcga tcaaacgcgc ggtggcgaat atggtggtca acgccgcccg ttatggcaat
     4561 ggctggatca aagtcagcag cggaacggag ccgaatcgcg cctggttcca ggtggaagat
     4621 gacggtccgg gaattgcgcc ggaacaacgt aagcacctgt tccagccgtt tgtccgcggc
     4681 gacagtgcgc gcaccattag cggcacggga ttagggctgg caattgtgca gcgtatcgtg
     4741 gataaccata acgggatgct ggagcttggc accagcgagc ggggcgggct ttccattcgc
     4801 gcctggctgc cagtgccggt aacgcgggcg cagggcacga caaaagaagg gtaatctaga
     4861 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     4921 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     4981 gcgtttatag gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gctt
//
