LOCUS       40924_6611        5531 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBL119-2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5531)
  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 5531)
  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 5531)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5531)
  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..5531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..6)
                     /label=AvrII Scar
                     /note="AvrII Scar"
     rep_origin      complement(151..739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(763..815)
                     /label=Tn3 Artifact
                     /note="Tn3 Artifact"
     misc_feature    complement(815..820)
                     /label=SpeI Scar
                     /note="SpeI Scar"
     terminator      complement(827..921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    complement(927..932)
                     /label=SacI Scar
                     /note="SacI Scar"
     CDS             complement(945..1601)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(1602..1704)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     misc_feature    1725..1730
                     /label=ClaI
                     /note="ClaI"
     regulatory      1730..1764
                     /label=J23114
                     /note="J23114"
                     /regulatory_class="promoter"
     regulatory      1774..1785
                     /label=B0034
                     /note="B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             1774..1785
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1793..2500
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    2504..2507
                     /label=Scar
                     /note="Scar"
     regulatory      2508..2559
                     /label=L3S1P52
                     /note="L3S1P52"
                     /regulatory_class="terminator"
     misc_feature    2560..2563
                     /label=Scar
                     /note="Scar"
     misc_feature    2585..2635
                     /label=E. coli LacZ Upstream
                     /note="E. coli LacZ Upstream"
     misc_feature    2637..2642
                     /label=XhoI Scar
                     /note="XhoI Scar"
     promoter        2643..2716
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     regulatory      2717..2747
                     /label=sRBS 3e2
                     /note="sRBS 3e2"
                     /regulatory_class="ribosome_binding_site"
     CDS             2748..3359
                     /codon_start=1
                     /product="ThsR"
                     /label=ThsR
                     /note="thsR"
                     /protein_id="AWU50953.1"
                     /translation="MQQQINGPVYLVDDDEAIIDSIDFLMEGYGYKLNSFNCGDRFLAE
                     VDLTQAGCVILDARMPGLTGPQVQQLLSDAKSPLAVIFLTGHGDVPMAVDAFKNGAFDF
                     FQKPVPGSLLSQSIAKGLTYSIDQHLKRTNQALIDTLSEREAQIFQLVIAGNTNKQMAN
                     ELCVAIRTIEVHRSKLMTKLGVNNLAELVKLAPLLAHKSE"
     misc_feature    2916..2918
                     /label=Phospho-asp
                     /note="Phospho-asp"
     terminator      3377..3448
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    3449..3456
                     /label=BB Scar
                     /note="BB Scar"
     terminator      3464..3491
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3498..3602
                     /label=Ptrp Fragment
                     /note="Ptrp Fragment"
     regulatory      3615..3806
                     /label=Promoter (Shal_3127)
                     /note="Promoter (Shal_3127)"
                     /regulatory_class="promoter"
     misc_feature    3667..3672
                     /label=half site
                     /note="half site"
     misc_feature    complement(3675..3680)
                     /label=half site
                     /note="half site"
     regulatory      3722..3727
                     /regulatory_class="minus_35_signal"
     regulatory      3742..3747
                     /regulatory_class="minus_10_signal"
     misc_feature    3754
                     /label=+1
                     /note="+1"
     regulatory      3807..3818
                     /label=B0034
                     /note="B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             3807..3818
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    3819..3824
                     /label=Scar
                     /note="Scar"
     CDS             3825..4538
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     regulatory      4557..4685
                     /label=B0015
                     /note="B0015"
                     /regulatory_class="terminator"
     regulatory      4557..4636
                     /label=B0010
                     /note="B0010"
                     /regulatory_class="terminator"
     terminator      4565..4636
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    4637..4644
                     /label=BB Scar
                     /note="BB Scar"
     regulatory      4645..4685
                     /label=B0012
                     /note="B0012"
                     /regulatory_class="terminator"
     terminator      4652..4679
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4686..4730
                     /label=pUC MCS
                     /note="pUC MCS"
     regulatory      4731..4765
                     /label=J23106
                     /note="J23106"
                     /regulatory_class="promoter"
     regulatory      4766..4799
                     /label=sRBS (50,000)
                     /note="sRBS (50,000)"
                     /regulatory_class="ribosome_binding_site"
     CDS             4800..5423
                     /label=TetR
                     /note="tetracycline repressor TetR"
     terminator      5432..5518
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 cctaggcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
       61 cacagaatca ggggataacg catgcaagaa catgtgagca aaaggccagc aaaaggccag
      121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
      601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      781 gaacgaaaac tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa
      841 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
      901 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
      961 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     1021 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1081 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1141 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     1201 aaataggcca ggttttcacc ataacacgcc acatcttgcg aatatatgtg tagaaactgc
     1261 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     1321 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     1381 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     1441 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     1501 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     1561 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     1621 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     1681 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgat ttatggctag
     1741 ctcagtccta ggtacaatgc tagcgaattc attaaagagg agaaaggtac ccatggtgag
     1801 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     1861 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1921 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     1981 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     2041 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2101 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     2161 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     2221 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2281 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     2341 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     2401 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     2461 gggccgccac tccaccggcg gcatggacga gctgtacaag taagctttct aactaaaaag
     2521 gcctcccaaa tcggggggcc ttttttattg ataacaaaac gctcgtctaa gaaaccatta
     2581 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg
     2641 agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacatca
     2701 gcaggacgca ctgaccttcc atatagcgta atctaaaaga ttacattatg cagcagcaaa
     2761 tcaacggccc ggtctacctg gtggatgatg atgaagccat tatcgactcc atcgattttt
     2821 tgatggaggg ctacggttac aaactgaact cgtttaactg cggcgatcgc tttttggcag
     2881 aagtcgatct cacccaggca ggatgtgtaa ttctggatgc gcgtatgcca ggcttaactg
     2941 gtcctcaggt gcaacagctg ctgagcgacg cgaaaagccc gcttgcggtc atcttcctga
     3001 ccggccatgg cgatgttccg atggcggttg atgcgttcaa aaatggcgcg ttcgatttct
     3061 ttcaaaaacc tgtgccgggt agcttgctca gtcagtcaat tgccaaaggc ttgacttatt
     3121 caatcgatca acatctgaaa cgtactaacc aagcgttaat cgacacgctc tcggaacgcg
     3181 aagctcaaat ttttcaactg gtgattgcag gcaacaccaa caaacagatg gctaacgagc
     3241 tttgcgtggc tattcgtacc attgaggttc accgtagcaa actgatgacc aaactgggtg
     3301 ttaacaacct ggctgaactg gttaaactgg cgccgctgct ggcacataaa tccgaataat
     3361 aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3421 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     3481 gcctttctgc gtttatagta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     3541 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     3601 tggttagcta gtcagcaaat ttgaactgtg aattgcttca caaacgccgc tatcgcaatg
     3661 tcagtatgtg gtttaccaca atatctaata tcactctgct caataacaat gatgaaaacc
     3721 ttaggaagaa gttaattgtg ttaaacagtt aactaggggc tttatctaac gctctcctaa
     3781 ggacaactgt cattgggaga tttaacaaag aggagaaata ctagatgcgt aaaggcgaag
     3841 agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata
     3901 agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt
     3961 tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt
     4021 atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt
     4081 ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt
     4141 acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga
     4201 aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa tacaatttta
     4261 acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta
     4321 aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca
     4381 ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg
     4441 ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg
     4501 cagcgggcat cacgcatggt atggatgaac tgtacaaatg atgataataa tctagaccag
     4561 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     4621 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     4681 ttataggtac ccgtggatcc tctagagtcg acctgcaggc atgcaagctt tttacggcta
     4741 gctcagtcct aggtatagtg ctagcccagc cagagaaaca ctctttaaca gggggtagta
     4801 tgatgtctcg tttagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg
     4861 tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta
     4921 cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt
     4981 tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac
     5041 gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac
     5101 atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt
     5161 tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgca gtggggcatt
     5221 ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa
     5281 cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc
     5341 aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac
     5401 aacttaaatg tgaaagtggg tcttaaggca tcaaataaaa cgaaaggctc agtcgaaaga
     5461 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     5521 gccgccctag a
//
