LOCUS       aldB-I_deletion_        4537 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 4537)
  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 4537)
  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 4537)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 20, 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..4537
                     /mol_type="other DNA"
                     /db_xref="taxon:2201328"
                     /organism="Cloning vector pK18sB"
     source          632..1404
                     /strain="KT2440"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160488"
                     /organism="Pseudomonas putida KT2440"
     source          1411..2175
                     /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..1404
                     /label=UsTR
     gene            complement(632..1167)
                     /locus_tag="PP_0544"
                     /label=PP_0544
     CDS             complement(632..1167)
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_0544"
                     /product="Ethanolamine transporter"
                     /label=PP_0544
                     /note="Evidence 1c : Function experimentally demonstratedin
                     the studied genus; PubMedId : 18833300; Product type t : 
                     transporter; Transport and binding proteins"
                     /protein_id="AAN66171.1"
                     /translation="MPSEHSAGSPAGSSVDFEKVGSDYFQQRELKKGAAGWVLLVGLGV
                     AYVISGDYAGWNFGLAQGGWGGMFLATLLMATMYLCMCFSLAELSSMIPTAGGGYGFAR
                     SAFGPWGGFLTGTAILIEYAIAPAAIAVFIGAYCQSLFGIGGWMIYLAFYIVFISIHIF
                     GVGEALKLMFIITAI"
     primer_bind     632..652
                     /label=oIAR001
                     /note="UpStream Targeting Region for AldB-I deletion"
     primer_bind     1379..1404
                     /label=overhang of oIAR003
                     /note="UsTR overhang of DsTR forward primer oIAR003"
     primer_bind     complement(1385..1404)
                     /label=oIAR002
                     /note="UpStream Targeting Region of aldB-I deletion"
     gene            complement(1395..1404)
                     /gene="aldB-I"
                     /locus_tag="PP_0545"
                     /label=aldB-I
     CDS             complement(1395..1404)
                     /codon_start=1
                     /transl_table=11
                     /gene="aldB-I"
                     /locus_tag="PP_0545"
                     /product="aldehyde dehydrogenase"
                     /EC_number="1.2.1.3"
                     /label=aldB-I
                     /note="Evidence 2a : Function of homologous 
                     geneexperimentally demonstrated in an other organism; 
                     PubMedId : 1732222; Product type e : enzyme; Energy 
                     metabolism"
                     /protein_id="AAN66172.1"
                     /translation="FF"
     misc_feature    1411..2175
                     /label=DsTR
     primer_bind     1411..1432
                     /label=oIAR003
                     /note="DownStream Targeting Region for AldB-I deletion"
     primer_bind     complement(1411..1432)
                     /label=overhang of oIAR002
                     /note="DsTR overhang of UsTR reverse primer oIAR002"
     gene            1664..2175
                     /locus_tag="PP_0546"
                     /label=PP_0546
     CDS             1664..2175
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_0546"
                     /product="sigma-54 dependent transcriptional regulator"
                     /label=PP_0546
                     /note="Evidence 2b : Function of strongly homologous gene; 
                     Product type r : regulator; Regulatory functions"
                     /protein_id="AAN66173.2"
                     /translation="MQSNHFTRHARQVHSVAHGGAGEGGSDPSIARSWLRCLEDYHLDP
                     AVIEAPVVLEHGRLLESRERLHQVLQIADHEMNSLHQQLSGAGHAVLLTDARGVILNCV
                     SAPAERRSFERAGLWLGADWSEAREGTNGIGTCLVERQALTIHQNEHFRGRHTGLTCSA
                     SPVFDPH"
     primer_bind     complement(2158..2175)
                     /label=oIAR004
                     /note="DownStream Targeting Region for AldB-I deletion"
     primer_bind     complement(2182..2205)
                     /label=DSTR primer overhang
                     /note="Where any DSTR constructed with a HindIII site can 
                     integrate into this plasmidPrimers that use this 
                     overhang:oIAR004; oIAR012"
     primer_bind     complement(2185..2202)
                     /label=M13F
                     /note="M13F"
     regulatory      2212..2261
                     /label=NEOKAN promoter
                     /note="NEOKAN promoter"
                     /regulatory_class="promoter"
     gene            2283..3077
                     /gene="nptII"
                     /label=nptII
     CDS             2283..3077
                     /codon_start=1
                     /transl_table=11
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="AWK60330.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3084..4505
                     /gene="sacB"
                     /label=sacB
     CDS             3084..4505
                     /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 cgcgatggcg gtgatgatga acatcagctt
      661 cagtgcctca cccacgccga aaatgtggat gctgatgaac acgatgtaga acgccaggta
      721 gatcatccag ccgccgatac cgaacagcga ttggcagtag gcgccgatga acaccgcgat
      781 ggcggctggg gcgatggcgt attcgatcag gatcgccgtg ccggtcaaaa agccgcccca
      841 cggtccgaac gcgctgcggg caaagccgta gccgccgcca gcggtgggga tcatcgacga
      901 cagttcggcc agcgagaagc acatgcacag gtacatggtt gccatcagca aggtggccag
      961 gaacatgccg ccccagccac cctgcgccag gccgaagttc cagccggcgt aatcgccgga
     1021 aatgacgtac gccacgccca ggccgaccag cagtacccag ccggcggcgc cttttttcag
     1081 ttcacgttgc tggaaatagt cagagccgac tttctcgaag tcgacggagg agcctgccgg
     1141 cgatccggcg gaatgttcgc ttggcataga ttcacctgtt ttttttcttg gtggccggaa
     1201 gggttccggg ccgatggtga tgggtgctgc aggaagcgag ccagagcggt tggtgcacgg
     1261 ttaccgccct gggttttgtg taaagcaagt ccggcctctt cgcgggcttg cccgctccca
     1321 caggtaccgc accggctctg gaaccaacgg tgaacctgta ggagcgggtt tacccgcgaa
     1381 gaaggcgacg cggtctagaa gaaggatatc gtgtgtctcc ttggtattgt gctttttagg
     1441 gagttgaaaa gcatagagca agggttgggc cagtgttggc ggtcccaggc aaatcaatga
     1501 cttgggtcat ttttacgggc ctgttcaagg accgctgtgc cggcgccggt acgcctgggg
     1561 tgacattttg tgccgtttgc gacacatcac aggcgtgtgc attgcaaagc ctaaccgggt
     1621 ggaggatgct gggctgacgc tctatctgcg gagaattaca acaatgcaga gcaatcactt
     1681 cactcgccat gcccggcagg tccatagcgt tgcccatggc ggggccgggg agggcggcag
     1741 tgatccgtcc atcgcccgtt cctggctgcg ctgcctggag gattaccacc tcgacccggc
     1801 ggtgatcgaa gcgccggtgg tgcttgagca tgggcgcctg ctggaaagcc gcgagcgcct
     1861 gcaccaggtg ctgcaaattg ccgaccatga aatgaacagc ctgcaccagc agctttccgg
     1921 cgcaggccac gcggtgctgc tgaccgacgc ccgcggggtg atcctcaatt gcgtcagtgc
     1981 ccccgccgag cggcgcagct tcgagcgtgc cgggctgtgg ctgggcgctg actggagcga
     2041 ggcgcgcgag ggcaccaatg gcatcggcac atgtctggtc gagcgccagg cgctgaccat
     2101 ccatcagaac gagcacttcc gtggccgtca caccgggctg acctgctcgg ccagcccggt
     2161 gttcgacccg catggaagct tggcactggc cgtcgtttta caacgtcgtg accggaattg
     2221 ccagctgggg cgccctctgg taaggttggg aagccctgca aacaggatga ggatcgtttc
     2281 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2341 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2401 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2461 tccaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2521 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2581 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2641 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2701 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2761 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg
     2821 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2881 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2941 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3001 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3061 ttgacgagtt cttctgagcg acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat
     3121 taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga
     3181 accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc
     3241 aaatccctga acagcaaaaa aatgaaaaat atcaagtttc tgaatttgat tcgtccacaa
     3301 ttaaaaatat ctcttctgca aaaggcctgg acgtttggga cagctggcca ttacaaaacg
     3361 ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc
     3421 ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc gaaacttcta
     3481 ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg
     3541 attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg
     3601 gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga
     3661 caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg
     3721 attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag cagttcatcg
     3781 atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag
     3841 ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag
     3901 gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag
     3961 aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc
     4021 tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg
     4081 catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat
     4141 ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg
     4201 atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca
     4261 aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac
     4321 acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca
     4381 gaggattcta cgcagacaaa caatcaacgt ttgcgccgag cttcctgctg aacatcaaag
     4441 gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta acagttaaca
     4501 aataatcaga ccccgtagaa aagatcaaag gatcttc
//
