LOCUS       pIAR003_1948_del        4498 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 4498)
  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 4498)
  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 4498)
  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..4498
                     /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          1388..2136
                     /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            632..1371
                     /locus_tag="PP_1947"
                     /label=PP_1947
     CDS             632..1371
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_1947"
                     /product="conserved protein of unknown function"
                     /label=PP_1947
                     /note="Evidence 4 : Homologs of previously reported genesof
                     unknown function"
                     /protein_id="AAN67563.1"
                     /translation="LTSTALAHAPITDLMIEHERRFLLMEEVTTAMLDKDGPAGADYHA
                     LNILGRKIADLFTNGKQVTVTCPNGTHLTASIEGRVGRAITGLPVVMRPGGGSGCAFPD
                     GETHVCPVEGTGEGTIVFDMTAHSVGALAEPITVTVKNGMAVSIEGGKQAAMWREIFAK
                     HGDPESFNCPAEIAIGLNPQVKQTGSMRTDKKMYATSHIGLGDTTALGGTCKAKLRLEG
                     VIREPCISVDGVVVTRGGKILID"
     primer_bind     632..651
                     /label=oIAR009
                     /note="UpStream Targeting Region F"
     primer_bind     complement(1351..1381)
                     /label=oIAR010
                     /note="UpStream Targeting Region R for 1948"
     primer_bind     1357..1381
                     /label=overhang of oIAR011
                     /note="UsTR overhang of DsTR forward primer oIAR011"
     misc_feature    1388..2136
                     /label=DsTR
     primer_bind     complement(1388..1411)
                     /label=overhang of oIAR010
                     /note="DsTR overhang of UsTR reverse primer oIAR010"
     primer_bind     1388..1407
                     /label=oIAR011
                     /note="DownStream Targeting Region F for 1948 deletion"
     gene            1409..2136
                     /locus_tag="PP_1949"
                     /label=PP_1949
     CDS             1409..2136
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="PP_1949"
                     /product="Oxidoreductase, GMC family"
                     /label=PP_1949
                     /protein_id="AAN67565.1"
                     /translation="MSEEQPVYDYIIIGAGSAGCVLANRLSANPEHSVLLLEAGSRPKG
                     LWASMPAGVSRVILPGPTNWAYQSEPDPSLAGRRIYVPRGKALGGSSAINGMAYLRGHR
                     EDYDHWVSLGCAGWGWDDVLPFYKKFEHREEGDEAFRGRDGELWVTDPVFKHPSSQAFI
                     ESCVEAGIPRLDDLNAPSPEGTGFLQFTIKGGRRHSAATAFLQPVLKRPNLHVLTGALV
                     QKIVIEAERATGVEYSLGNQ"
     primer_bind     complement(2116..2136)
                     /label=oIAR12
                     /note="DownStream Targeting Region R"
     primer_bind     complement(2143..2166)
                     /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(2146..2163)
                     /label=M13F
                     /note="M13F"
     regulatory      2173..2222
                     /label=NEOKAN promoter
                     /note="NEOKAN promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(2196..2214)
                     /label=oIAR008
                     /note="Reverse sequencing primer for targeting regions 
                     inserted at the HindIII site"
     gene            2244..3038
                     /gene="nptII"
                     /label=nptII
     CDS             2244..3038
                     /codon_start=1
                     /transl_table=11
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="AWK60330.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3045..4466
                     /gene="sacB"
                     /label=sacB
     CDS             3045..4466
                     /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 cacctgacga gtacagcact ggcacatgcc
      661 cccattaccg atctgatgat tgagcatgag cgccgtttct tgctcatgga ggaagtcacc
      721 acggccatgt tggataagga tggtccagcg ggtgctgatt accatgcgct gaatatcctc
      781 ggcaggaaga ttgctgatct tttcacaaac ggcaagcagg tcaccgttac ttgtccaaac
      841 ggtactcatt tgaccgccag tattgaaggt cgtgttgggc gtgccatcac tgggctacct
      901 gtcgttatgc ggccaggtgg cggtagtggc tgcgcatttc cggatggcga aacacacgtg
      961 tgcccggtcg aaggaaccgg cgagggcacg attgtttttg acatgaccgc tcactcggtg
     1021 ggtgccctag cggagcctat tacggtaacc gttaagaatg gcatggccgt gtctattgaa
     1081 ggtggcaagc aagcagctat gtggcgcgag attttcgcta agcacggcga ccctgaaagc
     1141 tttaattgtc cggctgaaat cgcgattggg ctcaatccgc aagttaaaca aacgggctcc
     1201 atgcgtaccg ataaaaaaat gtacgcaact tcacacattg gcctcggtga tacaacagcc
     1261 ctgggtggta cttgtaaggc gaagcttcga cttgaaggtg taattcggga gccctgcatc
     1321 agcgtcgatg gtgttgtggt aacccgtggt ggaaaaattt taatcgatta atcaaaagtg
     1381 cgatatcttc gggtgcgagc tgttgtgagt gagcgaggag cagccggtgt acgactacat
     1441 aatcattgga gcgggatctg ccggctgtgt tttggcaaac cggctttctg ccaacccaga
     1501 gcattcagtg ctactgctgg aagcgggttc acggccaaag ggactgtggg cttcgatgcc
     1561 agcaggggtg tcacgggtca tcctgcctgg gccgacgaat tgggcgtacc aatctgaacc
     1621 cgatcctagc ttggcagggc ggcgtatcta tgtgcctcgt ggaaaagcac tgggtggatc
     1681 gagtgcgatc aatgggatgg cttatcttcg aggccataga gaggactacg atcattgggt
     1741 atccctagga tgcgctgggt ggggttggga cgatgtcctg cctttctaca agaaatttga
     1801 acatcgggag gagggtgatg aagcattccg aggccgcgat ggcgagcttt gggtcacaga
     1861 cccggtattc aaacatccgt cctctcaggc gttcattgag tcttgtgtcg aagcgggaat
     1921 tcctcgtctg gacgatctca acgctccaag ccctgaaggc accggcttcc tgcaattcac
     1981 cattaaaggg gggcggcggc actcggcggc cactgcattc cttcagcctg tactgaagcg
     2041 cccaaacctg catgtgttga cgggcgcgct cgtccaaaaa atcgttattg aggctgagcg
     2101 tgccaccggt gtcgagtact ccctgggtaa tcagtcaagc ttggcactgg ccgtcgtttt
     2161 acaacgtcgt gaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     2221 aaacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2281 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2341 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2401 tgtccggtgc cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg ctggccacga
     2461 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2521 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2581 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2641 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2701 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2761 ggctcaaggc gcggatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     2821 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2881 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2941 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3001 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gacgatgaac atcaaaaagt
     3061 ttgcaaaaca agcaacagta ttaaccttta ctaccgcact gctggcagga ggcgcaactc
     3121 aagcgtttgc gaaagaaacg aaccaaaagc catataagga aacatacggc atttcccata
     3181 ttacacgcca tgatatgctg caaatccctg aacagcaaaa aaatgaaaaa tatcaagttt
     3241 ctgaatttga ttcgtccaca attaaaaata tctcttctgc aaaaggcctg gacgtttggg
     3301 acagctggcc attacaaaac gctgacggca ctgtcgcaaa ctatcacggc taccacatcg
     3361 tctttgcatt agccggagat cctaaaaatg cggatgacac atcgatttac atgttctatc
     3421 aaaaagtcgg cgaaacttct attgacagct ggaaaaacgc tggccgcgtc tttaaagaca
     3481 gcgacaaatt cgatgcaaat gattctatcc taaaagacca aacacaagaa tggtcaggtt
     3541 cagccacatt tacatctgac ggaaaaatcc gtttattcta cactgatttc tccggtaaac
     3601 attacggcaa acaaacactg acaactgcac aagttaacgt atcagcatca gacagctctt
     3661 tgaacatcaa cggtgtagag gattataaat caatctttga cggtgacgga aaaacgtatc
     3721 aaaatgtaca gcagttcatc gatgaaggca actacagctc aggcgacaac catacgctga
     3781 gagatcctca ctacgtagaa gataaaggcc acaaatactt agtatttgaa gcaaacactg
     3841 gaactgaaga tggctaccaa ggcgaagaat ctttatttaa caaagcatac tatggcaaaa
     3901 gcacatcatt cttccgtcaa gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg
     3961 ctgagttagc aaacggcgct ctcggtatga ttgagctaaa cgatgattac acactgaaaa
     4021 aagtgatgaa accgctgatt gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg
     4081 tctttaaaat gaacggcaaa tggtacctgt tcactgactc ccgcggatca aaaatgacga
     4141 ttgacggcat tacgtctaac gatatttaca tgcttggtta tgtttctaat tctttaactg
     4201 gcccatacaa gccgctgaac aaaactggcc ttgtgttaaa aatggatctt gatcctaacg
     4261 atgtaacctt tacttactca cacttcgctg tacctcaagc gaaaggaaac aatgtcgtga
     4321 ttacaagcta tatgacaaac agaggattct acgcagacaa acaatcaacg tttgcgccga
     4381 gcttcctgct gaacatcaaa ggcaagaaaa catctgttgt caaagacagc atccttgaac
     4441 aaggacaatt aacagttaac aaataatcag accccgtaga aaagatcaaa ggatcttc
//
