LOCUS       pIAR007_aldB-II_        4490 bp DNA     circular SYN 15-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 4490)
  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 4490)
  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 4490)
  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..4490
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          632..1377
                     /strain="KT2440"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160488"
                     /organism="Pseudomonas putida KT2440"
     source          1378..1383
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1378..1383
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          1384..2128
                     /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..1377
                     /label=UsTR
     gene            632..1225
                     /gene="qedH-II"
                     /locus_tag="PP_2679"
                     /label=qedH-II
     CDS             632..1225
                     /codon_start=1
                     /transl_table=11
                     /gene="qedH-II"
                     /locus_tag="PP_2679"
                     /product="quinoprotein ethanol dehydrogenase"
                     /EC_number="1.1.2.8"
                     /label=qedH-II
                     /note="Evidence 1b : Function experimentally demonstratedin
                     the studied species; PubMedId : 18218017, 7730276, 
                     19202108; Product type e : enzyme; Energy metabolism"
                     /protein_id="AAN68287.1"
                     /translation="SEAKGSSVFVAPAFLGAKNWMPMAYNKDTGLFYVPSNEWGMDIWN
                     EGIAYKKGAAFLGAGFTIKPLNEDYIGVLRAIDPVSGKEVWRHKNYAPLWGGVLTTKGN
                     LVFTGTPEGFLQAFNAKTGDKVWEFQTGSGVLGSPVTWEMDGEQYVSVVSGWGGAVPLW
                     GGEVAKRVKDFNQGGMLWTFKLPKQLQQTASVKP"
     primer_bind     632..649
                     /label=oIAR015
                     /note="UpStream Targeting Region, F"
     primer_bind     1357..1377
                     /label=overhang of oIAR017
                     /note="UsTR overhang of DsTR forward primer oIAR017"
     primer_bind     complement(1360..1377)
                     /label=oIAR016
                     /note="UpStream Targeting Region, R"
     misc_feature    1384..2128
                     /label=DsTR
     primer_bind     complement(1384..1404)
                     /label=overhang of oIAR016
                     /note="DsTR overhang of UsTR reverse primer oIAR016"
     primer_bind     1384..1401
                     /label=oIAR017
                     /note="DownStream Targeting Region, F"
     gene            1384
                     /gene="aldB-II"
                     /locus_tag="PP_2680"
                     /label=aldB-II
     CDS             1384
                     /codon_start=1
                     /transl_table=11
                     /gene="aldB-II"
                     /locus_tag="PP_2680"
                     /product="aldehyde dehydrogenase"
                     /EC_number="1.2.1.3"
                     /label=aldB-II (pedI in BioCyc)
                     /note="Evidence 2a : Function of homologous 
                     geneexperimentally demonstrated in an other organism; 
                     PubMedId : 7768815; Product type e : enzyme"
                     /protein_id="AAN68288.1"
                     /translation=""
     gene            1738..2010
                     /gene="pqqD-II"
                     /locus_tag="PP_2681"
                     /label=pqqD-II
     CDS             1738..2010
                     /codon_start=1
                     /transl_table=11
                     /gene="pqqD-II"
                     /locus_tag="PP_2681"
                     /product="coenzyme PQQ synthesis protein D"
                     /label=pqqD-II
                     /note="Evidence 2b : Function of strongly homologous gene; 
                     PubMedId : 2536663; Product type c : carrier"
                     /protein_id="AAN68289.1"
                     /translation="MNLIDRQQALALGRGLRLDWEPRKACHVLLYAGGIIELNASAGWV
                     LELLDGHSTVATVIDRLAQRFPNVPGLEEDVLAFLEVARAKSWIE"
     gene            2100..2128
                     /gene="yiaY"
                     /locus_tag="PP_2682"
                     /label=yiaY
     CDS             2100..2128
                     /codon_start=1
                     /transl_table=11
                     /gene="yiaY"
                     /locus_tag="PP_2682"
                     /product="Fe-containing alcohol dehydrogenase"
                     /EC_number="1.1.1.-"
                     /label=yiaY
                     /note="Evidence 3 : Function proposed based on presence of 
                     conserved amino acid motif, structural feature or limited 
                     homology; PubMedId : 2935393, 8185833; Product type e : 
                     enzyme; Central intermediary metabolism"
                     /protein_id="AAN68290.1"
                     /translation="MSQSFSPLR"
     primer_bind     complement(2108..2128)
                     /label=oIAR018
                     /note="DownStream Targeting Region, R"
     primer_bind     complement(2135..2158)
                     /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(2138..2155)
                     /label=M13F
                     /note="M13F"
     regulatory      2165..2214
                     /label=NEOKAN promoter
                     /note="NEOKAN promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(2188..2206)
                     /label=oIAR008
                     /note="Reverse sequencing primer for targeting regions 
                     inserted at the HindIII site"
     gene            2236..3030
                     /gene="nptII"
                     /label=nptII
     CDS             2236..3030
                     /codon_start=1
                     /transl_table=11
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="AWK60330.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3037..4458
                     /gene="sacB"
                     /label=sacB
     CDS             3037..4458
                     /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 cagcgaggcc aagggcagct cggtgttcgt
      661 cgcgccggcc ttcctcggcg ccaagaactg gatgccgatg gcctacaaca aggacacagg
      721 gctgttctac gtgccgtcca acgagtgggg catggacatc tggaacgaag gcatcgccta
      781 taagaaaggt gcggcgttcc tcggtgccgg cttcaccatc aagccgctca atgaagacta
      841 catcggcgtg ctgcgcgcca tcgacccggt cagcggcaag gaagtgtggc gccacaagaa
      901 ctatgcgccg ctgtggggcg gtgtgctgac caccaagggc aacctggtgt tcacgggcac
      961 gccagagggc ttcctgcagg cattcaacgc caagaccggc gacaaggtct gggaattcca
     1021 gaccggctcg ggcgtgctcg gctcgcccgt cacctgggaa atggacggcg agcaatacgt
     1081 ttcggtagtc tccggctggg gcggcgcggt gccgctgtgg ggcggcgaag tggccaaacg
     1141 ggtcaaggac ttcaaccagg gcggcatgct ctggaccttc aagttgccca agcagttgca
     1201 gcaaacggca agcgtcaagc cataacgcct gcctactgcc gcttgctgcc gctgatcggt
     1261 ggcaagcggc cctactacca aatgacgaga gccccgctgc caacgatgca ttcgcccggc
     1321 acgcggggct tcttaccatc ggtgcatcgc ctgtcgtggg acaggcaccg acccgcagat
     1381 atcaaccgcg tcgcggcctt cgcgggcttg cccgctccca caggttcacc gctggttcca
     1441 gaaccggtga agaacctgtg ggggcgggcg agcccgcaaa agggccggca catgcatggc
     1501 ctacccgtac caacgcaccc cgcaagctcg ttccaaagta gcattcggtc cagtccgccc
     1561 ccgtgcatta atgcttctac cggaatcctc cggcacaaga agaggacttg ccctatgtgg
     1621 actaaacccg cctataccga cctgcgcatt ggcttcgaag tgacgatgta tttcgccaac
     1681 cgttgatgac ggcccggcca tcgccacggt ggccggatcc tcctggagta tgcgcctgtg
     1741 aacctgattg atcgccagca ggcccttgcc ttggggcgag gcttgcgcct ggactgggag
     1801 ccacggaagg cttgccacgt gctgttgtac gccgggggca tcatcgagct caatgccagt
     1861 gccggctggg tcctcgagct gctggacggc cacagtaccg tggcaacggt catcgaccgc
     1921 ctggcacaac gcttccccaa tgtaccgggg ctcgaagagg acgtactggc gtttctggag
     1981 gtggcccgcg ccaaatcctg gatagaatga ccggtagccc cgctttggtc tggttgcttt
     2041 cgtggtggga tgctttacgc tggcggttat cttccagaac aataagaaca ggcttaccga
     2101 tgagccagag tttcagcccc cttcgcaaaa gcttggcact ggccgtcgtt ttacaacgtc
     2161 gtgaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaacagga
     2221 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2281 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2341 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2401 gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     2461 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2521 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     2581 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     2641 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     2701 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     2761 gcgcggatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     2821 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     2881 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     2941 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3001 ttctatcgcc ttcttgacga gttcttctga gcgacgatga acatcaaaaa gtttgcaaaa
     3061 caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt
     3121 gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc
     3181 catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt ttctgaattt
     3241 gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg
     3301 ccattacaaa acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca
     3361 ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc
     3421 ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa
     3481 ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca
     3541 tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc
     3601 aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc
     3661 aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta
     3721 cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct
     3781 cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa
     3841 gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca
     3901 ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta
     3961 gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg
     4021 aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa
     4081 atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc
     4141 attacgtcta acgatattta catgcttggt tatgtttcta attctttaac tggcccatac
     4201 aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc
     4261 tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc
     4321 tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc gagcttcctg
     4381 ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa
     4441 ttaacagtta acaaataatc agaccccgta gaaaagatca aaggatcttc
//
