LOCUS       40924_6616        5033 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBL132, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5033)
  AUTHORS   Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ.
  TITLE     Phosphatase activity tunes two-component system sensor detection 
            threshold
  JOURNAL   Nat Commun 9 (1), 1433 (2018)
  PUBMED    29650958
REFERENCE   2  (bases 1 to 5033)
  AUTHORS   Landry BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-OCT-2017) Bioengineering, Rice University, 6100 Main 
            St, Houston, TX 77005, USA
REFERENCE   3  (bases 1 to 5033)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5033)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2018"; volume: "9"; issue: "1"; pages: "1433"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-OCT-2017) Bioengineering, Rice University, 6100 Main St, 
            Houston, TX 77005, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..148
                     /label=PKanR from E. coli tn5
                     /note="PKanR from E. coli tn5"
                     /regulatory_class="promoter"
     CDS             149..937
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     terminator      952..1046
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1048..1049
                     /label=GC>CA
                     /note="GC>CA"
     rep_origin      1160..1705
                     /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."
     promoter        1878..1955
                     /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..3035
                     /label=lacI
                     /note="lac repressor"
     terminator      3044..3130
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    complement(3144..3149)
                     /label=AvrII Scar
                     /note="AvrII Scar"
     misc_feature    complement(3149..3196)
                     /label=ColE1 Fragment
                     /note="ColE1 Fragment"
     misc_feature    3258
                     /label=+1
                     /note="+1"
     promoter        3302..3330
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    3336
                     /label=+1
                     /note="+1"
     protein_bind    3338..3354
                     /label=lac operator
                     /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)."
     misc_feature    3364..3398
                     /label=sRBS 16573.91
                     /note="sRBS 16573.91"
     CDS             3399..4853
                     /codon_start=1
                     /product="Taz"
                     /label=Taz
                     /protein_id="AWU50960.1"
                     /translation="MINRIRVVTLLVMVLGVFALLQLISGSLFFSSLHHSQKSFVVSNQ
                     LREQQGELTSTWDLMLQTRINLSRSAVRMMMDSSNQQSNAKVELLDSARKTLAQAATHY
                     KKFKSMAPLPEMVATSRNIDEKYKNYYTALTELIDYLDYGNTGAYFAQPTQGMQNAMGE
                     AFAQYALSSEKLYRDIVTDNADDYRFAQWQLAVIALVVVLILLVAWYGIRRMLLTPLAK
                     IIAHIREIAGGNLANTLTIDGRSEMGDLAQSVSHMAAGVKQLADDRTLLMAGVSHDLRT
                     PLTRIRLATEMMSEQDGYLAESINKDIEECNAIIEQFIDYLRTGQEMPMEMADLNAVLG
                     EVIAAESGYEREIETALYPGSIEVKMHPLSIKRAVANMVVNAARYGNGWIKVSSGTEPN
                     RAWFQVEDDGPGIAPEQRKHLFQPFVRGDSARTISGTGLGLAIVQRIVDNHNGMLELGT
                     SERGGLSIRAWLPVPVTRAQGTTKEG"
     misc_feature    3399..4166
                     /label=Tar
                     /note="Tar"
     misc_feature    4167..4850
                     /label=Cytosolic Domain
                     /note="Cytosolic Domain"
     misc_feature    4167..4367
                     /label=A Domain
                     /note="A Domain"
     misc_feature    4167..4187
                     /label=HAMP
                     /note="HAMP"
     misc_feature    4182..4283
                     /label=alpha 1
                     /note="alpha 1"
     misc_feature    4197..4379
                     /label=HisKA
                     /note="HisKA"
     misc_feature    4227..4244
                     /label=H-Box
                     /note="H-Box"
     misc_feature    4227..4229
                     /label=Phospho Acceptor
                     /note="Phospho Acceptor"
     misc_feature    4248..4253
                     /label=SDAA
                     /note="SDAA"
     misc_feature    4260..4265
                     /label=SDAA
                     /note="SDAA"
     misc_feature    4269..4274
                     /label=SDAA
                     /note="SDAA"
     misc_feature    4287..4367
                     /label=alpha 2
                     /note="alpha 2"
     misc_feature    4305..4307
                     /label=SDAA
                     /note="SDAA"
     misc_feature    4314..4319
                     /label=SDAA
                     /note="SDAA"
     misc_feature    4368..4850
                     /label=B Domain
                     /note="B Domain"
     misc_feature    4383..4403
                     /label=beta 1
                     /note="beta 1"
     misc_feature    4404..4424
                     /label=alpha 3
                     /note="alpha 3"
     misc_feature    4434..4469
                     /label=beta 2
                     /note="beta 2"
     misc_feature    4470..4484
                     /label=beta 3
                     /note="beta 3"
     misc_feature    4488..4556
                     /label=alpha 4
                     /note="alpha 4"
     misc_feature    4494..4817
                     /label=HATPase
                     /note="HATPase"
     misc_feature    4527..4544
                     /label=N-box
                     /note="N-box"
     misc_feature    4566..4586
                     /label=beta 4
                     /note="beta 4"
     misc_feature    4596..4616
                     /label=beta 5
                     /note="beta 5"
     misc_feature    4605..4634
                     /label=D-Box
                     /note="D-Box"
     misc_feature    4659..4670
                     /label=F-Box
                     /note="F-Box"
     misc_feature    complement(4686..4703)
                     /label=Priming Region
                     /note="Priming Region"
     misc_feature    4701..4718
                     /label=G2-box
                     /note="G2-box"
     misc_feature    4701..4715
                     /label=GXGXG Motif
                     /note="GXGXG Motif"
     misc_feature    4710..4745
                     /label=alpha 5
                     /note="alpha 5"
     misc_feature    4758..4781
                     /label=beta 6
                     /note="beta 6"
     misc_feature    4788..4808
                     /label=beta 7
                     /note="beta 7"
     terminator      complement(4871..4914)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    4940..4947
                     /label=BB Scar
                     /note="BB Scar"
     terminator      4955..4982
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4989..5033
                     /label=pUC19 MCS
                     /note="pUC19 MCS"
ORIGIN
        1 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
       61 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
      121 tcaagagaca ggatgaggat cgtttcgcat 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 tcaaccacca
     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 agacctaggc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     3241 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     3301 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
     3361 acaagaacac ttagagaaca aacgggagag tatctaaaat gattaaccgt atccgcgtag
     3421 tcacgctgtt ggtaatggtg ctgggggtat tcgcactgtt acagcttatt tccggcagtc
     3481 tgtttttttc ttcccttcac catagccaga agagctttgt ggtttccaat caattacggg
     3541 aacagcaggg cgagctgacg tcaacctggg atttaatgct gcaaacgcgc attaacctga
     3601 gtcgttcagc ggtacggatg atgatggatt cctccaatca acaaagtaac gccaaagttg
     3661 aattgctcga tagcgccagg aaaacattgg cgcaggcagc gacgcattat aaaaaattca
     3721 aaagcatggc accgttacct gaaatggtcg ctaccagtcg taatattgat gaaaaatata
     3781 aaaactatta cacagcgtta actgaactga ttgattatct agattatggc aatactggag
     3841 cttatttcgc tcagccaacc cagggaatgc aaaatgcaat gggcgaagcg tttgctcagt
     3901 acgccctcag cagtgaaaaa ctgtatcgcg atatcgtcac tgacaacgca gatgattacc
     3961 gatttgccca gtggcaactg gcggttatcg cgctggtggt ggtattgatt ctgctggtgg
     4021 cgtggtacgg cattcgccgt atgttgctta ctccgctggc aaaaattatt gctcacattc
     4081 gcgaaatcgc cggtggtaac ctggcgaata ccctgaccat tgacgggcgc agtgaaatgg
     4141 gcgacctggc gcagagcgtt tcacatatgg cggctggtgt taagcaactg gcggatgacc
     4201 gcacgctgct gatggcgggg gtaagtcacg acttgcgcac gccgctgacg cgtattcgcc
     4261 tggcgactga gatgatgagc gagcaggatg gctatctggc agaatcgatc aataaagata
     4321 tcgaagagtg caacgccatc attgagcagt ttatcgacta cctgcgcacc gggcaggaga
     4381 tgccgatgga aatggcggat cttaatgcag tactcggtga ggtgattgct gccgaaagtg
     4441 gctatgagcg ggaaattgaa accgcgcttt accccggcag cattgaagtg aaaatgcacc
     4501 cgctgtcgat caaacgcgcg gtggcgaata tggtggtcaa cgccgcccgt tatggcaatg
     4561 gctggatcaa agtcagcagc ggaacggagc cgaatcgcgc ctggttccag gtggaagatg
     4621 acggtccggg aattgcgccg gaacaacgta agcacctgtt ccagccgttt gtccgcggcg
     4681 acagtgcgcg caccattagc ggcacgggat tagggctggc aattgtgcag cgtatcgtgg
     4741 ataaccataa cgggatgctg gagcttggca ccagcgagcg gggcgggctt tccattcgcg
     4801 cctggctgcc agtgccggta acgcgggcgc agggcacgac aaaagaaggg taatctagac
     4861 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4921 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     4981 cgtttatagg tacccgggga tcctctagag tcgacctgca ggcatgcaag ctt
//
