LOCUS       40924_26596        5596 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKD227, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5596)
  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 5596)
  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 5596)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5596)
  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..5596
                     /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..3397
                     /label=sRBS (1096.98)
                     /note="sRBS (1096.98)"
     CDS             3398..5413
                     /codon_start=1
                     /product="Sbal195_3859 (TtrS)"
                     /label=Sbal195_3859
                     /protein_id="AWU50966.1"
                     /translation="MFSSGMSLKWPLSIQGSIEPKMTPRIIFRMALRTKVFITLLACLF
                     ASANVFAVEPQNQSAPSDVMDAEVVAPVATEEPGYQVVDVGVLAIRGYKATINRWQPLM
                     GWLETQIPNSYFRLHPLSLDELAKGVETQGLDFVITNPGQSVLLARQYSLTWLATLRSP
                     LNNGAAMQVGSALVVRADSPYQTLTDLKGRRLGIVSKNAFGGYLTLVYEAQLKGIDLPR
                     FVGEIIPLGFPLDNLLYQLDDQKSGNDAAKDNRLDATVVPVCQLEQMQAEGLINIGHYR
                     VLDNQTPVGFHCQVSTRLYPNWSMAKTNRASQSLAKSVTQALLALPEDHLAAKSSDSAG
                     WTTAVSQLAIDQLLKDLDMHPLQTPWWQRAWQWVKLHQQWAWFILGILVLLNAYHFWLE
                     YRFSRRGRELINTQRQLNENRALLEHAQRIAIAGELGASLSHELNQPLAAIGHYCHGAE
                     VRLQRGTSPEELQSVLTLIQQEVTRADSIISRLRNLLKKRPVSKQALYLHELVNETVPL
                     LAYEFEQHQINLAVNVSGEPYLQSLDEVGMQQLLLNLLKNAFDACVQRLELESSGTEQS
                     GMRKPYVPTIDIDLRYQERTLLLTVTDNGTGLTEETSLLMQAFYSTKSEGLGLGLVICR
                     DIAESHGGTFSLESAMGGGCQAQVAIPRKPEPNGVL"
     misc_feature    3503..3557
                     /label=TM
                     /note="TM"
     regulatory      4490..4499
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4532..4588
                     /label=TM
                     /note="TM"
     misc_feature    4718..4720
                     /label=Catalytic His
                     /note="Catalytic His"
     terminator      complement(5434..5477)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    5503..5510
                     /label=BB Scar
                     /note="BB Scar"
     terminator      5518..5545
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    5552..5596
                     /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 acatatacct cttacactac tatataaatt ttcttccatg ttttcttccg gcatgagcct
     3421 gaaatggcct ctgagcatcc agggttccat cgaaccgaaa atgactccgc gcatcatctt
     3481 ccgcatggcc ctgcgcacta aagtgttcat caccctgctg gcttgcctgt ttgcttctgc
     3541 caacgtattc gcggtggaac cgcagaacca gtccgcgcct tccgatgtga tggacgctga
     3601 ggtggttgct ccggttgcga cggaggaacc gggttaccag gtcgttgatg ttggtgtgct
     3661 ggcgattcgc ggttataaag ctaccatcaa ccgttggcag ccactgatgg gttggctgga
     3721 gactcagatc ccgaactcct acttccgcct gcacccgctg tctctggacg agctggcaaa
     3781 gggtgtagaa acccagggtc tggacttcgt aatcaccaat ccaggccagt ccgtactgct
     3841 ggcgcgtcag tattctctga cgtggctggc gaccctgcgc tctccgctga acaacggtgc
     3901 tgcgatgcag gtgggttccg cgctggtagt tcgtgccgac agcccgtacc agaccctgac
     3961 cgatctgaaa ggccgtcgtc tgggcatcgt cagcaaaaac gcattcggtg gttatctgac
     4021 gctggtatat gaggcacagc tgaaaggcat cgatctgccg cgctttgttg gtgaaatcat
     4081 cccactgggc ttcccgctgg ataacctgct gtatcagctg gatgatcaga aatctggtaa
     4141 tgacgccgca aaagacaatc gtctggatgc caccgttgta ccggtatgcc aactggaaca
     4201 gatgcaggcc gaaggtctga tcaacatcgg ccactatcgc gttctggaca accagacccc
     4261 ggtaggtttc cactgtcaag taagcactcg tctgtatccg aactggtcta tggctaaaac
     4321 taaccgtgcc agccagtccc tggcaaaatc tgtgacccag gctctgctgg ctctgccgga
     4381 agatcacctg gcagcgaaaa gctctgactc tgcaggttgg accactgcag tgtcccagct
     4441 ggcgatcgac cagctgctga aagacctgga catgcatcca ctgcagaccc catggtggca
     4501 gcgcgcttgg cagtgggtaa aactgcacca gcagtgggcg tggttcattc tgggtatcct
     4561 ggtcctgctg aacgcatacc atttctggct ggaataccgt ttctcccgtc gtggtcgtga
     4621 actgatcaac acccagcgcc agctgaacga aaaccgtgct ctgctggagc acgcacaacg
     4681 tatcgccatc gctggcgaac tgggtgctag cctgtcccat gaactgaacc agccgctggc
     4741 tgcaattggt cactattgcc atggtgcgga agtgcgtctg cagcgtggta cttctcctga
     4801 agaactgcag tctgttctga ccctgatcca gcaggaagtt acgcgcgccg actctatcat
     4861 cagccgtctg cgtaacctgc tgaaaaaacg cccggttagc aaacaggctc tgtatctgca
     4921 cgaactggta aacgaaacgg tgccgctgct ggcgtacgaa ttcgaacagc accagatcaa
     4981 cctggcggta aacgtcagcg gtgaacctta cctgcaatct ctggacgagg tcggtatgca
     5041 gcagctgctg ctgaacctgc tgaaaaacgc gtttgacgcg tgcgttcagc gcctggaact
     5101 ggaatcctcc ggcactgaac agagcggtat gcgtaaaccg tatgttccta cgatcgatat
     5161 tgatctgcgc taccaggagc gtactctgct gctgactgtg acggataacg gcactggtct
     5221 gaccgaggag actagcctgc tgatgcaggc cttctattcc accaaatctg aaggcctggg
     5281 cctgggtctg gttatttgcc gtgatatcgc tgagtcccac ggtggcacct tctctctgga
     5341 atccgccatg ggtggtggct gccaggctca ggtggccatc ccgcgcaagc cggaaccgaa
     5401 tggcgttctg taataatcta gaccaggcat caaataaaac gaaaggctca gtcgaaagac
     5461 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     5521 tcaccttcgg gtgggccttt ctgcgtttat aggtacccgg ggatcctcta gagtcgacct
     5581 gcaggcatgc aagctt
//
