LOCUS       40924_26586        5364 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKD182, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5364)
  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 5364)
  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 5364)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5364)
  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..5364
                     /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"
     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..3396
                     /label=sRBS (1051.22)
                     /note="sRBS (1051.22)"
     CDS             3397..5181
                     /codon_start=1
                     /product="Shal_3128"
                     /label=Shal_3128
                     /protein_id="AWU50963.1"
                     /translation="MSRLLLCICVLLFSSVAWSKPQQFYVGVLANWGHQQAVERWTPMM
                     EYLNEHVPDAEFHVYPGNFKALNLAMELGQIQFIITNPGQYLYLSNQYPLSWLATMRSK
                     RHDGTTSAIGSAIIVRADSDYRTLYDLKGKVVAASDPHALGGYQATVGLMHSLGMDPDT
                     FFGETKFLGFPLDPLLYQVRDGNVDAAITPLCTLEDMVARGVLKSSDFRVLNPSRPDGV
                     ECQCSTTLYPNWSFAATESVSTELSKEITQALLELPSDSPAAIKAQLTGWTSPISQLAV
                     IKLFKELHVKTPDSSRWEAVKKWLEENRHWGILSVLVFIIATLYHLWIEYRFHQKSSSL
                     IESERQLKQQAVALERLQSASIVGEIGAGLAHEINQPIAAITSYSEGGIMRLQGKEQAD
                     TDSCIELLEKIHKQSTRAGEVVHRIRGLLKRREAVMVDVNILTLVEESISLLRLELARR
                     EIQINTQIKGEPFFITADRVGLLQVLINLIKNSLDAIAESDNARSGKINIELDFKEYQV
                     NVSIIDNGPGLAMDSDTLMATFYTTKMDGLGLGLAICREVISNHDGHFLLSNRDDGVLG
                     CVATLNLKKRGSEVPIEV"
     misc_feature    4237
                     /label=Mutation
                     /note="Mutation"
     misc_feature    4483..4539
                     /label=H-Box
                     /note="H-Box"
     misc_feature    4510..4512
                     /label=Catalytic His
                     /note="Catalytic His"
     misc_feature    4531..4536
                     /label=SDR
                     /note="SDR"
     misc_feature    4543..4548
                     /label=SDR
                     /note="SDR"
     misc_feature    4552..4557
                     /label=SDR
                     /note="SDR"
     misc_feature    4621..4623
                     /label=SDR
                     /note="SDR"
     misc_feature    4630..4635
                     /label=SDR
                     /note="SDR"
     misc_feature    4822..4881
                     /label=N-Box
                     /note="N-Box"
     misc_feature    4951..4980
                     /label=D-Box
                     /note="D-Box"
     misc_feature    complement(5006..5034)
                     /label=Priming Region
                     /note="Priming Region"
     misc_feature    5032..5145
                     /label=G-Box
                     /note="G-Box"
     misc_feature    5032..5046
                     /label=GxGxG Motif
                     /note="GxGxG Motif"
     misc_feature    5035..5037
                     /label=Mutantation Point
                     /note="Mutantation Point"
     misc_feature    5038..5058
                     /label=Priming Region
                     /note="Priming Region"
     terminator      complement(5202..5245)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    5271..5278
                     /label=BB Scar
                     /note="BB Scar"
     terminator      5286..5313
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    5320..5364
                     /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 acaaacgacc atcgacgtcc gaccctaacg tatactatgt cccgcctgct gctgtgtatc
     3421 tgtgttctgc tgttctcttc tgtggcgtgg tctaaaccgc agcagtttta tgtgggcgta
     3481 ctggctaact ggggtcatca gcaagccgtt gaacgttgga ccccgatgat ggagtatctg
     3541 aacgaacatg tgccggacgc ggaatttcac gtctacccgg gcaacttcaa agcactgaac
     3601 ctggcaatgg aactgggcca gattcagttc attatcacta acccgggcca atatctgtac
     3661 ctgagcaatc agtacccgct gtcttggctg gcgaccatgc gttctaagcg tcacgatggt
     3721 accacttctg cgatcggttc cgccattatt gtccgcgcgg acagcgacta ccgcaccctg
     3781 tacgacctga aaggtaaagt ggtggctgcg tccgacccgc atgctctggg tggctaccaa
     3841 gcgaccgtcg gtctgatgca ttccctgggc atggatccgg acaccttctt cggtgaaacc
     3901 aagtttctgg gctttccact ggatccgctg ctgtaccaag ttcgtgatgg caacgttgac
     3961 gcggccatta ccccactgtg cactctggag gacatggttg cacgcggcgt actgaaatct
     4021 tccgattttc gtgtgctgaa ccctagccgc ccggatggtg tagaatgcca gtgctctacc
     4081 accctgtacc cgaactggtc tttcgctgcg actgagtctg tatccaccga actgtctaaa
     4141 gaaatcacgc aggcactgct ggaactgcca tccgacagcc cggcagctat caaagcgcaa
     4201 ctgaccggct ggaccagccc gatctcccaa ctggcggtaa tcaaactgtt caaagagctg
     4261 cacgtaaaaa ccccggactc tagccgttgg gaagccgtta agaagtggct ggaagaaaac
     4321 cgtcactggg gtatcctgtc tgttctggtg ttcatcattg caacgctgta tcacctgtgg
     4381 attgaatacc gcttccacca aaaaagctct tctctgatcg aatctgaacg tcagctgaaa
     4441 cagcaagctg ttgccctgga acgtctgcaa tctgctagca tcgttggtga aattggtgcg
     4501 ggtctggccc acgagattaa tcagccgatc gctgcaatta cctcttattc tgaaggtggc
     4561 atcatgcgcc tgcaaggtaa agaacaggcg gatacggata gctgcatcga actgctggaa
     4621 aaaatccaca aacagagcac tcgcgcaggc gaagtggtgc accgcatccg tggtctgctg
     4681 aaacgtcgtg aagcggtgat ggtagatgtt aacatcctga ccctggtgga agaatccatc
     4741 agcctgctgc gtctggagct ggcacgtcgc gaaatccaga tcaacactca gatcaaaggt
     4801 gaaccgttct tcattactgc cgaccgcgtt ggcctgctgc aagttctgat taacctgatc
     4861 aaaaactccc tggacgcgat cgctgaatct gataatgccc gttctggtaa aatcaacatc
     4921 gaactggact ttaaagagta ccaggtaaac gtctccatca tcgataacgg tccgggcctg
     4981 gcgatggatt ctgacactct gatggctacg ttttacacta ccaaaatgga tggcctgggc
     5041 ctgggtctgg caatctgccg cgaagttatc agcaaccacg acggccactt cctgctgtcc
     5101 aaccgtgacg acggcgttct gggctgtgtg gcaaccctga atctgaaaaa acgcggttct
     5161 gaagtgccga tcgaagtcta ataatctaga ccaggcatca aataaaacga aaggctcagt
     5221 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     5281 acactggctc accttcgggt gggcctttct gcgtttatag gtacccgggg atcctctaga
     5341 gtcgacctgc aggcatgcaa gctt
//
