LOCUS       pIAR015_shunt_de        4499 bp DNA     circular SYN 06-JUN-2022
DEFINITION  Cloning vector pK18sB, complete sequence.
ACCESSION   MH166772
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4499)
  AUTHORS   Jayakody LN, Johnson CW, Whitham JM, Giannone RJ, Black BA, 
            Cleveland NS, Klingeman DM, Michener WE, Olstad JL, Vardon DR, Brown
            RC, Brown SD, Hettich RL, Guss AM, Beckham GT.
  TITLE     Thermochemical wastewater valorization via enhanced microbial 
            toxicity tolerance
  JOURNAL   Energy Environ Sci (2018) In press
REFERENCE   2  (bases 1 to 4499)
  AUTHORS   Johnson CW.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2018) National Bioenergy Center, National 
            Renewable Energy Laboratory, 15301 Denver West Parkway, Golden, CO 
            80401, USA
REFERENCE   3  (bases 1 to 4499)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 22, 2024 from SnapGene 7.1.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Energy 
            Environ Sci (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-APR-2018) National Bioenergy Center, National Renewable Energy 
            Laboratory, 15301 Denver West Parkway, Golden, CO 80401, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          632..1381
                     /strain="KT2440"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160488"
                     /organism="Pseudomonas putida KT2440"
     source          1388..2137
                     /strain="KT2440"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160488"
                     /organism="Pseudomonas putida KT2440"
     rep_origin      1..595
                     /label=pMB1 replication origin
                     /note="pMB1 replication origin"
     primer_bind     509..528
                     /label=oIAR007
                     /note="Forward sequencing primer for targeting regions 
                     inserted at the EcoRI site"
     primer_bind     601..617
                     /note="special M13R binding site; M13R"
     primer_bind     602..625
                     /label=USTR primer overhang
                     /note="Where any USTR constructed with an EcoRI site can 
                     integrate into this plasmidPrimers that use this 
                     overhang:oIAR001"
     gene            632..1381
                     /locus_tag="PP_3353"
                     /label=PP_3353
     CDS             632..1381
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_3353"
                     /product="putative sulfatase-modifying factor 
                     (C-alpha-formyglycine- generating enzyme)"
                     /label=PP_3353
                     /note="Evidence 3 : Function proposed based on presence of 
                     conserved amino acid motif, structural feature or limited 
                     homology; Product type pe : putative enzyme; 
                     Unknownfunction : Enzymes of unknown specificity"
                     /protein_id="AAN68957.2"
                     /translation="FARFVKATGYVTHAERGTRVEDDPALPDALRIPGAMVFHQGADVL
                     GPGWQFVPGANWRHPQGPGSSLAGLDNHPVVQIALEDAQAYARWAGRELPSEAQLEYAM
                     RGGLTDADFSWGTTEQPKGKLMANTWQGQFPYRNAAKDGFTGTSPVGCFPANGFGLFDA
                     GGNVWELTRTGYRPGHDAQRDAKLDPSGPALSDSFDPADPGVPVAVIKGGSHLCSADRC
                     MRYRPSARQPQPVFMTTSHVGFRTIRQ"
     misc_feature    632..1381
                     /label=UsTR
     primer_bind     632..649
                     /label=oIAR121
                     /note="UsTR annealing portion F"
     primer_bind     1357..1381
                     /label=overhang of oIAR123
     primer_bind     complement(1359..1381)
                     /label=oIAR122
                     /note="UsTR annealing R"
     gene            complement(1362..1381)
                     /locus_tag="PP_3354"
                     /label=PP_3354
     CDS             complement(1362..1381)
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_3354"
                     /product="putative Acyl-CoA dehydrogenase, ferrulic acid 
                     biotransformation protein"
                     /label=PP_3354
                     /note="Evidence 3 : Function proposed based on presence of 
                     conserved amino acid motif, structural feature or limited 
                     homology"
                     /protein_id="AAN68958.1"
                     /translation="IAESF"
     misc_feature    1388..2137
                     /label=DsTR
     primer_bind     complement(1388..1411)
                     /label=overhang of oIAR122
     primer_bind     1388..1406
                     /label=oIAR123
     gene            complement(1435..2137)
                     /gene="fcs"
                     /locus_tag="PP_3356"
                     /label=fcs
     CDS             complement(1435..2137)
                     /codon_start=1
                     /transl_table=11
                     /gene="fcs"
                     /locus_tag="PP_3356"
                     /product="feruloyl-CoA-synthetase"
                     /EC_number="6.2.1.34"
                     /label=fcs
                     /note="Evidence 1b : Function experimentally demonstrated
                     in the studied species; PubMedId : 12534466, 10543794, 
                     9579070, 10222033; Product type e : enzyme; 
                     Biologicalprocesses : Scavenge (Catabolism)"
                     /protein_id="AAN68960.2"
                     /translation="TTGPLSMAGYVGLPAPGCEVKLVPVGDKLEARFRGPHIMPGYWRS
                     PQQTAEAFDEEGFYCSGDALKLADARQPELGLMFDGRIAEDFKLSSGVFVSVGPLRNRA
                     VLEGSPYVQDIVVTAPDRECLGLLVFPRLPECRRLAGLAEDASDARVLANDTVRSWFAD
                     WLERLNRDAQGNASRIEWLSLLAEPPSIDAGEITDKGSINQRAVLQRRAAQVEALYRGE
                     DPDALHAKVRP"
     primer_bind     complement(2119..2137)
                     /label=oIAR124
     primer_bind     complement(2144..2167)
                     /label=DSTR primer overhang
                     /note="Where any DSTR constructed with a HindIII site can 
                     integrate into this plasmidPrimers that use this 
                     overhang:oIAR004"
     primer_bind     complement(2147..2164)
                     /label=M13F
                     /note="M13F"
     regulatory      2174..2223
                     /label=NEOKAN promoter
                     /note="NEOKAN promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(2197..2215)
                     /label=oIAR008
                     /note="Reverse sequencing primer for targeting regions 
                     inserted at the HindIII site"
     gene            2245..3039
                     /gene="nptII"
                     /label=nptII
     CDS             2245..3039
                     /codon_start=1
                     /transl_table=11
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="AWK60330.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3046..4467
                     /gene="sacB"
                     /label=sacB
     CDS             3046..4467
                     /codon_start=1
                     /transl_table=11
                     /gene="sacB"
                     /product="levan sucrase"
                     /label=sacB
                     /protein_id="AWK60331.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgcctcaca
      601 caggaaacag ctatgacatg attacgaatt cttcgcgcgc ttcgtcaagg ccacggggta
      661 tgtcacccat gccgagcgcg gtacccgtgt cgaggacgac cctgccctgc ccgacgcgct
      721 gcggataccg ggtgcgatgg tgtttcatca gggtgcggac gtgctcggcc ccggctggca
      781 gttcgtgccc ggcgccaact ggcgacaccc gcaagggccg ggcagcagcc tggccgggct
      841 ggacaaccat ccggtggtgc agatcgccct ggaagatgcc caggcctatg cccgctgggc
      901 aggccgcgaa ctgcccagcg aggcgcagct ggaatacgcc atgcgcggcg gcctgaccga
      961 tgccgacttc agctggggta ccaccgagca gcccaagggc aagctcatgg ccaatacctg
     1021 gcagggtcag ttcccttatc gcaatgcggc gaaggatggt tttaccggta catcgcccgt
     1081 gggttgcttc ccggccaacg gctttggcct gttcgatgcc ggcggcaatg tctgggagct
     1141 gactcgcacg ggctatcggc caggccatga cgcacagcgc gacgccaagc tcgacccctc
     1201 aggcccggcc ctgagtgaca gcttcgaccc ggcagacccc ggcgtgccgg tggcggtaat
     1261 caaaggcggc tcgcacctgt gttcggcgga ccgctgcatg cgctaccgcc cctcggcacg
     1321 ccagccgcag ccggtgttca tgacgacctc gcacgtgggt ttcagaacga ttcggcaatg
     1381 agatatcctg gacgatggcc gtgtcctcga agacggtgca cagcccggcc gcactcaagg
     1441 ccgcaccttg gcgtgcaatg cgtcggggtc ttcgccacgg tacagcgcct cgacctgagc
     1501 ggcgcgccgc tgcagcacgg cgcgctgatt gatcgagccc ttgtcggtga tttcaccggc
     1561 gtcgatcgac ggcggctcgg ccagcagcga cagccattcg atacggctgg cgttgccttg
     1621 ggcatcgcgg ttcaagcgct ccagccagtc agcgaaccaa ctgcgcacgg tgtcgttggc
     1681 cagcacccgc gcatcgctgg catcctctgc cagcccggcc aggcgccgac actcgggcag
     1741 acgcgggaac accagcaggc ccaggcattc acggtccggc gcggtgacca cgatgtcctg
     1801 tacgtaaggc gagccctcca gcactgcgcg gttgcgcagc ggcccgacac tgacgaatac
     1861 cccggacgaa agtttgaagt cctcagcgat acggccatcg aacatcaggc caagctcggg
     1921 ctgcctggca tcggccagct tcaacgcgtc gcccgaacag tagaagccct cctcgtcgaa
     1981 cgcctcggcg gtctgctgcg gcgagcgcca gtagcccggc atgatatgcg ggccacggaa
     2041 gcgcgcctcg agcttgtcgc ccaccggcac cagcttcact tcgcagccag gtgccggcag
     2101 cccgacatag ccggccatcg acaaaggccc ggtggtgaag cttggcactg gccgtcgttt
     2161 tacaacgtcg tgaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     2221 caaacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2281 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2341 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2401 ctgtccggtg ccctgaatga actccaagac gaggcagcgc ggctatcgtg gctggccacg
     2461 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2521 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2581 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2641 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2701 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2761 aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2821 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2881 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2941 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3001 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgacgatgaa catcaaaaag
     3061 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
     3121 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
     3181 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt
     3241 tctgaatttg attcgtccac aattaaaaat atctcttctg caaaaggcct ggacgtttgg
     3301 gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg ctaccacatc
     3361 gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta catgttctat
     3421 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
     3481 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
     3541 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
     3601 cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc agacagctct
     3661 ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg aaaaacgtat
     3721 caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa ccatacgctg
     3781 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
     3841 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
     3901 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
     3961 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
     4021 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
     4081 gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc aaaaatgacg
     4141 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
     4201 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
     4261 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
     4321 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgccg
     4381 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
     4441 caaggacaat taacagttaa caaataatca gaccccgtag aaaagatcaa aggatcttc
//
