LOCUS       PP457286                3282 bp    DNA     circular SYN 16-MAR-2024
DEFINITION  Cloning vector pSpyC, complete sequence.
ACCESSION   PP457286
VERSION     PP457286.1
KEYWORDS    .
SOURCE      Cloning vector pSpyC
  ORGANISM  Cloning vector pSpyC
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 3282)
  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  JOURNAL   Unpublished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 3282)
  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..3282
                     /organism="Cloning vector pSpyC"
                     /mol_type="other DNA"
                     /db_xref="taxon:3129133"
     regulatory      56..142
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      157..197
                     /regulatory_class="promoter"
                     /note="BG42 synthetic bacterial promoter (Zobel et al.,
                     2015); BG42"
     misc_RNA        249..324
                     /note="tracrRNA scaffold for sgRNA; gRNA scaffold"
     regulatory      326..369
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      462..556
                     /regulatory_class="terminator"
                     /note="transcription terminator t0 from phage lambda; t0"
     CDS             688..1503
                     /note="confers resistance to kanamycin in bacteria; KmR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="WWZ17848.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1670..1778
                     /note="incP origin of transfer"
     CDS             complement(1789..2451)
                     /note="replication protein for the broad host range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /codon_start=1
                     /transl_table=11
                     /product="pBBR1 Rep"
                     /protein_id="WWZ17849.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL
                     HHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAY
                     VVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGP
                     GEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     rep_origin      2452..3223
                     /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 aattaagccc attgacaagg ctctcgcggc
      181 caggtataat tgcacgacct aggccgcggc cgcgcgaatt cgagctcggt ctcatcacta
      241 gggtctctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      301 gaaaaagtgg caccgagtcg gtgctataaa acgaaaggcc cagtctttcg actgagcctt
      361 tcgttttata actaggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg
      421 cggccgcgtc gtgactggga aaaccctggc gactagtctt ggactcctgt tgatagatcc
      481 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
      541 ttttattggt gagaatccag gggtccccaa taattacgat ttaaatttgt gtctcaaaat
      601 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
      661 tacataaaca gtaatacaag gggtgttatg agccatattc agcgtgaaac gagctgtagc
      721 cgtccgcgtc tgaacagcaa catggatgcg gatctgtatg gctataaatg ggcgcgtgat
      781 aacgtgggtc agagcggcgc gaccatttat cgtctgtatg gcaaaccgga tgcgccggaa
      841 ctgtttctga aacatggcaa aggcagcgtg gcgaacgatg tgaccgatga aatggtgcgt
      901 ctgaactggc tgaccgaatt tatgccgctg ccgaccatta aacattttat tcgcaccccg
      961 gatgatgcgt ggctgctgac caccgcgatt ccgggcaaaa ccgcgtttca ggtgctggaa
     1021 gaatatccgg atagcggcga aaacattgtg gatgcgctgg ccgtgtttct gcgtcgtctg
     1081 catagcattc cggtgtgcaa ctgcccgttt aacagcgatc gtgtgtttcg tctggcccag
     1141 gcgcagagcc gtatgaacaa cggcctggtg gatgcgagcg attttgatga tgaacgtaac
     1201 ggctggccgg tggaacaggt gtggaaagaa atgcataaac tgctgccgtt tagcccggat
     1261 agcgtggtga cccacggcga ttttagcctg gataacctga ttttcgatga aggcaaactg
     1321 attggctgca ttgatgtggg ccgtgtgggc attgcggatc gttatcagga tctggccatt
     1381 ctgtggaact gcctgggcga atttagcccg agcctgcaaa aacgtctgtt tcagaaatat
     1441 ggcattgata atccggatat gaacaaactg caatttcatc tgatgctgga tgaatttttc
     1501 taataattaa ttggaccgcg gtccgcgcgt tgtccttttc cgctgcataa ccctgcttcg
     1561 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
     1621 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
     1681 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
     1741 cggtgctcaa cgggaatcct gctctgcgag gctggccgta ggccggccct accggcgcgg
     1801 cagcgttacc cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac acggctcatc
     1861 gggcatcggc aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg tcaaggctgg
     1921 caggtctggt tccatgcccg gaatgccggg ctggctgggc ggctcctcgc cggggccggt
     1981 cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc catgccccaa
     2041 cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca ccgacaggcg
     2101 caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg ccgacacggt
     2161 gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt ccttgactgc
     2221 gtattggacc gtccgcaaag aacgtccgat gagcttggaa agtgtcttct ggctgaccac
     2281 cacggcgttc tggtggccca tctgcgccac gaggtgatgc agcagcattg ccgccgtggg
     2341 tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt gcacccagtg
     2401 accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca tgcttatctc
     2461 catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca acttttcggc
     2521 agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt ttctttaacc
     2581 tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa
     2641 gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg tgcccaaaga
     2701 agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg
     2761 gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc
     2821 ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc
     2881 ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg
     2941 ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac cggcaagcga
     3001 agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa ttgatggggc
     3061 aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg ggtagccggt
     3121 gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc ttgtccagca
     3181 gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc catcgtccac
     3241 atatccacgg gctggcaagg gagcgcagcg accgcgcagg gc
//

