LOCUS       pIAR008_amaB_del        4506 bp DNA     circular SYN 22-JUN-2020
DEFINITION  Cloning vector pK18sB, complete sequence.
ACCESSION   MH166772
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  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 4506)
  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 4506)
  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..4506
                     /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          1382..1387
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1382..1387
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1382..1387
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1382..1387
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1382..1387
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1382..1387
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1388..2144
                     /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"
     misc_feature    632..1381
                     /label=UsTR
     gene            complement(632..1236)
                     /gene="amaA"
                     /locus_tag="PP_5257"
                     /label=amaA
     CDS             complement(632..1236)
                     /codon_start=1
                     /transl_table=11
                     /gene="amaA"
                     /locus_tag="PP_5257"
                     /product="L-pipecolate oxidase"
                     /EC_number="1.5.3.7"
                     /label=amaA
                     /note="Evidence 2a : Function of homologous 
                     geneexperimentally demonstrated in an other organism; 
                     PubMedId : 16237033, 18974823; Product type e : enzyme; 
                     Energymetabolism : Amino acids and amines"
                     /protein_id="AAN70822.2"
                     /translation="MEPAMSELRQQCLWEHVSKPTVAAQALAGEHKADVCVIGGGITGL
                     SAAIHLLEQGKSVIVLEAWKIGHGGSGRNVGLVNAGTWIRPDDVEATLGQKQGSRLNKV
                     LGEAPAEVFAMIERLGIDCQAQHKGTLHMAHNATGIADLEARHEQWRRRGADVELLTGA
                     QCQEYCGTDKISAALLDRRAGTINPMGYTQGLAAAVTR"
     primer_bind     632..650
                     /label=oIAR033
                     /note="UpStream Targeting Region, F"
     primer_bind     1357..1381
                     /label=overhang of oIAR035
                     /note="UsTR overhang of DsTR forward primer"
     primer_bind     complement(1361..1381)
                     /label=oIAR034
                     /note="UpStream Targeting Region, R"
     misc_feature    1388..2144
                     /label=DsTR
     primer_bind     complement(1388..1414)
                     /label=overhang of oIAR034
                     /note="DsTR overhang of UsTR reverse primer"
     primer_bind     1388..1409
                     /label=oIAR035
                     /note="DownStream Targeting Region, F"
     gene            1523..2144
                     /locus_tag="PP_5259"
                     /label=PP_5259
     CDS             1523..2144
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_5259"
                     /product="Transcriptional regulator, LysR family"
                     /label=PP_5259
                     /protein_id="AAN70824.1"
                     /translation="MSKRLVPSMTALQCFEAAARHLSFTRAAEELHLTQSAVSKQVAQL
                     EDMLRHHLFLRIRRRLQLTPAGSLYLAEVNKILTQVDMSSRYVLTYGEQTEILKVATQP
                     SFGVRWLIPHLKGFGKRHGNIHLDIRNEMEPFALLQGSADVVFFYGQGTWPGATCVELF
                     REEVVPVCAPELLAGRKLAGAEELAELVLLQSTSRPEAWHEWFL"
     primer_bind     complement(2125..2144)
                     /label=oIAR036
                     /note="DownStream Targeting Region, R"
     primer_bind     complement(2151..2174)
                     /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(2154..2171)
                     /label=M13F
                     /note="M13F"
     regulatory      2181..2230
                     /label=NEOKAN promoter
                     /note="NEOKAN promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(2204..2222)
                     /label=oIAR008
                     /note="Reverse sequencing primer for targeting regions 
                     inserted at the HindIII site"
     gene            2252..3046
                     /gene="nptII"
                     /label=nptII
     CDS             2252..3046
                     /codon_start=1
                     /transl_table=11
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="AWK60330.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3053..4474
                     /gene="sacB"
                     /label=sacB
     CDS             3053..4474
                     /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 caggcgcgtt acagcagccg ccaggccctg
      661 ggtgtagccc atggggttga tggtgccagc acggcgatca agcagcgcag cggaaatctt
      721 gtcggtaccg cagtattcct ggcactgggc accggtcagc agctcgacgt cggcacctcg
      781 gcggcgccat tgttcgtgac gcgcttcgag gtcggcgata ccggtggcgt tgtgcgccat
      841 gtgcagcgtg cctttgtgct gggcctggca gtcgatgccg aggcgctcga tcatggcgaa
      901 cacttcagcc ggggcttcgc caagcacctt gttcaggcgg ctaccctgct tctggccaag
      961 ggtggcctcg acgtcgtcgg ggcggatcca cgtgccggcg ttgaccaggc cgacgttgcg
     1021 cccggagcca ccgtggccga tcttccaggc ctccagcacg atcaccgact tgccctgttc
     1081 gagcaggtgg atggccgccg acaggccggt aataccgccg ccgatcacac agacatcggc
     1141 cttgtgctca ccggccaggg cctgggcggc gacggtcggt ttgctgacgt gttcccacaa
     1201 gcattgttga cgcagttcgg acatggccgg ctccagcaat ttgttcttgt tggggccgca
     1261 gggcggccct ttcgcgggta aacccgctcc tacaggggtc ggtgtcgctt ggacgaaccc
     1321 tgtaggagcg ggtctacccg cgaaccgggg gccaggcccc cggccatgca gcatcaatac
     1381 agatatcttg tgcatctcct acatcggtga tggggtggaa aaatgatgtg gatcagtatt
     1441 aatccgatca cataggtaca acaaacgacc tttattccgc atatcattcc gtcaggtaat
     1501 gatttgagcc gcagagaccg ctatgtcaaa acgcctggta ccctccatga ccgccctgca
     1561 gtgctttgaa gctgcagccc gccatctgag ttttacccgt gccgccgaag aactgcacct
     1621 gacccagagc gcggtcagca agcaggtggc gcagctggaa gacatgctgc gccaccacct
     1681 gttcctgcgc atccgccggc gcttgcagct gaccccggcc ggcagcctgt acctggccga
     1741 ggtcaacaaa atcctgaccc aggtggatat gtccagccgc tatgtgctca cctacggcga
     1801 gcagaccgag atattgaagg tagccactca accgagtttt ggcgtgcgct ggttgatccc
     1861 gcacctcaag gggtttggca aacggcacgg caatatccac ctggacattc gcaacgagat
     1921 ggagccgttc gccctgctgc aaggctcggc tgacgtggtg ttcttctatg gccagggcac
     1981 ctggccgggg gccacttgcg tggagttgtt tcgcgaggag gtggtgccgg tgtgtgcgcc
     2041 agagctgctg gccgggcgca agctggccgg tgccgaggag ctggccgagc tggtgctgct
     2101 gcaaagcacc tcgcgcccgg aggcctggca tgagtggttc ctggaagctt ggcactggcc
     2161 gtcgttttac aacgtcgtga ccggaattgc cagctggggc gccctctggt aaggttggga
     2221 agccctgcaa acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2281 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2341 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2401 gaccgacctg tccggtgccc tgaatgaact ccaagacgag gcagcgcggc tatcgtggct
     2461 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2521 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2581 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2641 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2701 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2761 gttcgccagg ctcaaggcgc ggatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2821 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2881 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2941 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3001 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcga cgatgaacat
     3061 caaaaagttt gcaaaacaag caacagtatt aacctttact accgcactgc tggcaggagg
     3121 cgcaactcaa gcgtttgcga aagaaacgaa ccaaaagcca tataaggaaa catacggcat
     3181 ttcccatatt acacgccatg atatgctgca aatccctgaa cagcaaaaaa atgaaaaata
     3241 tcaagtttct gaatttgatt cgtccacaat taaaaatatc tcttctgcaa aaggcctgga
     3301 cgtttgggac agctggccat tacaaaacgc tgacggcact gtcgcaaact atcacggcta
     3361 ccacatcgtc tttgcattag ccggagatcc taaaaatgcg gatgacacat cgatttacat
     3421 gttctatcaa aaagtcggcg aaacttctat tgacagctgg aaaaacgctg gccgcgtctt
     3481 taaagacagc gacaaattcg atgcaaatga ttctatccta aaagaccaaa cacaagaatg
     3541 gtcaggttca gccacattta catctgacgg aaaaatccgt ttattctaca ctgatttctc
     3601 cggtaaacat tacggcaaac aaacactgac aactgcacaa gttaacgtat cagcatcaga
     3661 cagctctttg aacatcaacg gtgtagagga ttataaatca atctttgacg gtgacggaaa
     3721 aacgtatcaa aatgtacagc agttcatcga tgaaggcaac tacagctcag gcgacaacca
     3781 tacgctgaga gatcctcact acgtagaaga taaaggccac aaatacttag tatttgaagc
     3841 aaacactgga actgaagatg gctaccaagg cgaagaatct ttatttaaca aagcatacta
     3901 tggcaaaagc acatcattct tccgtcaaga aagtcaaaaa cttctgcaaa gcgataaaaa
     3961 acgcacggct gagttagcaa acggcgctct cggtatgatt gagctaaacg atgattacac
     4021 actgaaaaaa gtgatgaaac cgctgattgc atctaacaca gtaacagatg aaattgaacg
     4081 cgcgaacgtc tttaaaatga acggcaaatg gtacctgttc actgactccc gcggatcaaa
     4141 aatgacgatt gacggcatta cgtctaacga tatttacatg cttggttatg tttctaattc
     4201 tttaactggc ccatacaagc cgctgaacaa aactggcctt gtgttaaaaa tggatcttga
     4261 tcctaacgat gtaaccttta cttactcaca cttcgctgta cctcaagcga aaggaaacaa
     4321 tgtcgtgatt acaagctata tgacaaacag aggattctac gcagacaaac aatcaacgtt
     4381 tgcgccgagc ttcctgctga acatcaaagg caagaaaaca tctgttgtca aagacagcat
     4441 ccttgaacaa ggacaattaa cagttaacaa ataatcagac cccgtagaaa agatcaaagg
     4501 atcttc
//
