LOCUS       PP457287                3269 bp    DNA     circular SYN 16-MAR-2024
DEFINITION  Cloning vector pSpyD, complete sequence.
ACCESSION   PP457287
VERSION     PP457287.1
KEYWORDS    .
SOURCE      Cloning vector pSpyD
  ORGANISM  Cloning vector pSpyD
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 3269)
  AUTHORS   Roghair Stroud,M.N., Vang,D.X. and Halverson,L.J.
  TITLE     Design and optimization of a robust CRISPR interference system for
            use in Pseudomonas putida KT2440
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3269)
  AUTHORS   Roghair Stroud,M.N., Vang,D.X. and Halverson,L.J.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAR-2024) Plant Pathology, Entomology, and
            Microbiology, Iowa State University, 2213 Pammel Dr, Ames, IA
            50011, USA
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3269
                     /organism="Cloning vector pSpyD"
                     /mol_type="other DNA"
                     /db_xref="taxon:3129135"
     regulatory      56..142
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      157..184
                     /regulatory_class="promoter"
                     /note="BG28 synthetic bacterial promoter (Zobel et al.,
                     2015); BG28"
     misc_RNA        236..311
                     /note="tracrRNA scaffold for sgRNA; gRNA scaffold"
     regulatory      313..356
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      449..543
                     /regulatory_class="terminator"
                     /note="transcription terminator t0 from phage lambda; t0"
     CDS             675..1490
                     /note="confers resistance to kanamycin in bacteria; KmR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="WWZ17850.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1657..1795
                     /note="incP origin of transfer"
     CDS             complement(1776..2438)
                     /note="replication protein for the broad host range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /codon_start=1
                     /transl_table=11
                     /product="pBBR1 Rep"
                     /protein_id="WWZ17851.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL
                     HHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAY
                     VVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGP
                     GEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     rep_origin      2439..3210
                     /note="replication origin for the broad host range plasmid
                     pBBR1 from Bordetella bronchiseptica, requires the pBBR1
                     Rep protein for replication"
ORIGIN      
        1 gaagcccgga gagcaagccc gtaggggggg cgcgcccagc tgtctagggc ggcggatttg
       61 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
      121 actgagcctt tcgttttatt tgatgccttt aattaactag gttgacatgg atataatgta
      181 tgtacctagg ccgcggccgc gcgaattcga gctcggtctc atcactaggg tctctgtttt
      241 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
      301 cgagtcggtg ctataaaacg aaaggcccag tctttcgact gagcctttcg ttttataact
      361 aggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttgcgg ccgcgtcgtg
      421 actgggaaaa ccctggcgac tagtcttgga ctcctgttga tagatccagt aatgacctca
      481 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      541 aatccagggg tccccaataa ttacgattta aatttgtgtc tcaaaatctc tgatgttaca
      601 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
      661 atacaagggg tgttatgagc catattcagc gtgaaacgag ctgtagccgt ccgcgtctga
      721 acagcaacat ggatgcggat ctgtatggct ataaatgggc gcgtgataac gtgggtcaga
      781 gcggcgcgac catttatcgt ctgtatggca aaccggatgc gccggaactg tttctgaaac
      841 atggcaaagg cagcgtggcg aacgatgtga ccgatgaaat ggtgcgtctg aactggctga
      901 ccgaatttat gccgctgccg accattaaac attttattcg caccccggat gatgcgtggc
      961 tgctgaccac cgcgattccg ggcaaaaccg cgtttcaggt gctggaagaa tatccggata
     1021 gcggcgaaaa cattgtggat gcgctggccg tgtttctgcg tcgtctgcat agcattccgg
     1081 tgtgcaactg cccgtttaac agcgatcgtg tgtttcgtct ggcccaggcg cagagccgta
     1141 tgaacaacgg cctggtggat gcgagcgatt ttgatgatga acgtaacggc tggccggtgg
     1201 aacaggtgtg gaaagaaatg cataaactgc tgccgtttag cccggatagc gtggtgaccc
     1261 acggcgattt tagcctggat aacctgattt tcgatgaagg caaactgatt ggctgcattg
     1321 atgtgggccg tgtgggcatt gcggatcgtt atcaggatct ggccattctg tggaactgcc
     1381 tgggcgaatt tagcccgagc ctgcaaaaac gtctgtttca gaaatatggc attgataatc
     1441 cggatatgaa caaactgcaa tttcatctga tgctggatga atttttctaa taattaattg
     1501 gaccgcggtc cgcgcgttgt ccttttccgc tgcataaccc tgcttcgggg tcattatagc
     1561 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     1621 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     1681 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     1741 gaatcctgct ctgcgaggct ggccgtaggc cggccctacc ggcgcggcag cgttacccgt
     1801 gtcggcggct ccaacggctc gccatcgtcc agaaaacacg gctcatcggg catcggcagg
     1861 cgctgctgcc cgcgccgttc ccattcctcc gtttcggtca aggctggcag gtctggttcc
     1921 atgcccggaa tgccgggctg gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc
     1981 tcgcccggat acagggtcgg gatgcggcgc aggtcgccat gccccaacag cgattcgtcc
     2041 tggtcgtcgt gatcaaccac cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg
     2101 ggctggcccc acgccacgcg gtcattgacc acgtaggccg acacggtgcc ggggccgttg
     2161 agcttcacga cggagatcca gcgctcggcc accaagtcct tgactgcgta ttggaccgtc
     2221 cgcaaagaac gtccgatgag cttggaaagt gtcttctggc tgaccaccac ggcgttctgg
     2281 tggcccatct gcgccacgag gtgatgcagc agcattgccg ccgtgggttt cctcgcaata
     2341 agcccggccc acgcctcatg cgctttgcgt tccgtttgca cccagtgacc gggcttgttc
     2401 ttggcttgaa tgccgatttc tctggactgc gtggccatgc ttatctccat gcggtagggg
     2461 tgccgcacgg ttgcggcacc atgcgcaatc agctgcaact tttcggcagc gcgacaacaa
     2521 ttatgcgttg cgtaaaagtg gcagtcaatt acagattttc tttaacctac gcaatgagct
     2581 attgcggggg gtgccgcaat gagctgttgc gtacccccct tttttaagtt gttgattttt
     2641 aagtctttcg catttcgccc tatatctagt tctttggtgc ccaaagaagg gcacccctgc
     2701 ggggttcccc cacgccttcg gcgcggctcc ccctccggca aaaagtggcc cctccggggc
     2761 ttgttgatcg actgcgcggc cttcggcctt gcccaaggtg gcgctgcccc cttggaaccc
     2821 ccgcactcgc cgccgtgagg ctcggggggc aggcgggcgg gcttcgccct tcgactgccc
     2881 ccactcgcat aggcttgggt cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct
     2941 gcgcgagcct tgacccgcct tccacttggt gtccaaccgg caagcgaagc gcgcaggccg
     3001 caggccggag gcttttcccc agagaaaatt aaaaaaattg atggggcaag gccgcaggcc
     3061 gcgcagttgg agccggtggg tatgtggtcg aaggctgggt agccggtggg caatccctgt
     3121 ggtcaagctc gtgggcaggc gcagcctgtc catcagcttg tccagcaggg ttgtccacgg
     3181 gccgagcgaa gcgagccagc cggtggccgc tcgcggccat cgtccacata tccacgggct
     3241 ggcaagggag cgcagcgacc gcgcagggc
//

