LOCUS       40924_26601        5437 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKD233.7-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5437)
  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 5437)
  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 5437)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5437)
  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..5437
                     /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"
     regulatory      1730..1764
                     /label=constitutive promoter J23114
                     /note="constitutive promoter J23114"
                     /regulatory_class="promoter"
     regulatory      1772..1785
                     /label=RBS pprotet
                     /note="RBS pprotet"
                     /regulatory_class="ribosome_binding_site"
     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..2751
                     /label=sRBS (1087.31)
                     /note="sRBS (1087.31)"
                     /regulatory_class="ribosome_binding_site"
     CDS             2752..3372
                     /codon_start=1
                     /product="Sbal195_3858 (TtrR)"
                     /label=Sbal195_3858
                     /protein_id="AWU50974.1"
                     /translation="MSLALPVYLIDDDDSVRRSLRFMLESYGLKITDFDSAEAFFTAVD
                     LTLPGCALVDVRMPGLSGPQLHLELVSKNSPLAVIYLTGHGDVPMAVEALKLGAVDFFQ
                     KPADGAKLAEAVVKALEHAKTHYQDNQYLETYQALTPREREILNLIAQGLKNQEIADNL
                     CIAMRTVEIHRANLMKGMQVGSLAEMMLIYSRIAERLRLEDNR"
     misc_feature    2914..2916
                     /label=D55
                     /note="D55"
     terminator      3390..3461
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    3462..3469
                     /label=BB Scar
                     /note="BB Scar"
     terminator      3477..3504
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3511..3615
                     /label=Ptrp Fragment
                     /note="Ptrp Fragment"
     regulatory      3628..3712
                     /label=Psbal195_3860 (185-269)
                     /note="Psbal195_3860 (185-269)"
                     /regulatory_class="promoter"
     misc_feature    3630..3653
                     /label=Actual repeat
                     /note="Actual repeat"
     misc_feature    3630..3634
                     /label=Repeat
                     /note="Repeat"
     misc_feature    3635..3648
                     /label=Fnr
                     /note="Fnr"
     misc_feature    3642..3653
                     /label=Repeat
                     /note="Repeat"
     misc_feature    complement(3646..3650)
                     /label=Repeat
                     /note="Repeat"
     regulatory      3650..3655
                     /regulatory_class="minus_35_signal"
     regulatory      3676..3681
                     /regulatory_class="minus_10_signal"
     misc_feature    3688
                     /label=+1
                     /note="+1"
     misc_feature    3697..3703
                     /label=Repeat
                     /note="Repeat"
     misc_feature    complement(3709..3715)
                     /label=Repeat
                     /note="Repeat"
     regulatory      3713..3730
                     /label=BBa_B0034
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             3713..3724
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3731..4444
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     regulatory      4463..4591
                     /label=B0015
                     /note="B0015"
                     /regulatory_class="terminator"
     regulatory      4463..4542
                     /label=B0010
                     /note="B0010"
                     /regulatory_class="terminator"
     terminator      4471..4542
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    4543..4550
                     /label=BB Scar
                     /note="BB Scar"
     regulatory      4551..4591
                     /label=B0012
                     /note="B0012"
                     /regulatory_class="terminator"
     terminator      4558..4585
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4592..4636
                     /label=pUC MCS
                     /note="pUC MCS"
     regulatory      4637..4671
                     /label=constitutive promoter J23106
                     /note="constitutive promoter J23106"
                     /regulatory_class="promoter"
     regulatory      4672..4705
                     /label=sRBS (50,000)
                     /note="sRBS (50,000)"
                     /regulatory_class="ribosome_binding_site"
     CDS             4706..5329
                     /label=TetR
                     /note="tetracycline repressor TetR"
     terminator      5338..5424
                     /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 ctgaccactt taattcatta ccacacttaa taggagcgat aatgagcctg
     2761 gcgctgccgg tgtacctgat tgacgacgat gactccgttc gccgttccct gcgtttcatg
     2821 ctggaaagct acggcctgaa aatcactgac ttcgattctg ctgaagcgtt cttcaccgcg
     2881 gtagacctga ccctgccggg ttgcgcactg gtggacgtac gtatgccggg cctgagcggc
     2941 ccacagctgc acctggaact ggtgtctaag aacagcccgc tggccgtgat ctatctgacc
     3001 ggtcacggcg acgttccgat ggcggttgaa gcgctgaaac tgggtgcggt agatttcttt
     3061 cagaaacctg cagacggcgc taaactggct gaagctgtgg tcaaagccct ggaacacgcg
     3121 aaaacccact accaagacaa ccagtacctg gaaacctatc aggctctgac cccacgtgag
     3181 cgcgaaatcc tgaacctgat tgcgcaaggt ctgaaaaacc aggaaatcgc ggacaacctg
     3241 tgcattgcga tgcgcaccgt agaaatccac cgtgcgaacc tgatgaaagg tatgcaggtg
     3301 ggcagcctgg cagagatgat gctgatttac tctcgtatcg cggagcgtct gcgcctggaa
     3361 gataaccgtt aataatctag accaggcatc aaataaaacg aaaggctcag tcgaaagact
     3421 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     3481 caccttcggg tgggcctttc tgcgtttata gtaaatcact gcataattcg tgtcgctcaa
     3541 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
     3601 ttctgaaatg agctggttag ctagtcaata tttgttgcgc tagatcaaat ccacgcctga
     3661 tatgtggaaa accactatag ttatgccgcc tcgcctttta cagaatgcct gtaaagagga
     3721 gaaatactag atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga
     3781 actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc
     3841 aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg
     3901 gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca
     3961 tatgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac
     4021 gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga
     4081 taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct
     4141 gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca
     4201 aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca
     4261 gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga
     4321 caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca
     4381 tatggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta
     4441 caaatgatga taataatcta gaccaggcat caaataaaac gaaaggctca gtcgaaagac
     4501 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     4561 tcaccttcgg gtgggccttt ctgcgtttat aggtacccgt ggatcctcta gagtcgacct
     4621 gcaggcatgc aagcttttta cggctagctc agtcctaggt atagtgctag cccagccaga
     4681 gaaacactct ttaacagggg gtagtatgat gtctcgttta gataaaagta aagtgattaa
     4741 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
     4801 ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt
     4861 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
     4921 aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact
     4981 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
     5041 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt
     5101 atatgcactc agcgcagtgg ggcattttac tttaggttgc gtattggaag atcaagagca
     5161 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
     5221 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
     5281 attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtctt aaggcatcaa
     5341 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     5401 aacgctctcc tgagtaggac aaatccgccg ccctaga
//
