LOCUS       PP457289                4033 bp    DNA     circular SYN 16-MAR-2024
DEFINITION  Cloning vector pSpyB-GFP, complete sequence.
ACCESSION   PP457289
VERSION     PP457289.1
KEYWORDS    .
SOURCE      Cloning vector pSpyB-GFP
  ORGANISM  Cloning vector pSpyB-GFP
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4033)
  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  JOURNAL   Unpublished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 4033)
  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..4033
                     /organism="Cloning vector pSpyB-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:3129132"
     regulatory      56..142
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      157..197
                     /regulatory_class="promoter"
                     /note="BG35 synthetic bacterial promoter (Zobel et al.,
                     2015); BG35"
     gene            263..991
                     /gene="msfgfp"
     CDS             263..991
                     /gene="msfgfp"
                     /note="monomeric superfolder green fluorescent protein"
                     /codon_start=1
                     /transl_table=11
                     /product="msfGFP"
                     /protein_id="WWZ17855.1"
                     /translation="MIMGIHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNG
                     KLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI
                     SFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADK
                     QKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYK"
     misc_RNA        1000..1075
                     /note="tracrRNA scaffold for sgRNA; gRNA scaffold"
     regulatory      1077..1120
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      1213..1307
                     /regulatory_class="terminator"
                     /note="transcription terminator t0 from phage lambda; t0"
     CDS             1439..2254
                     /note="confers resistance to kanamycin in bacteria; KmR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="WWZ17856.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2421..2529
                     /note="incP origin of transfer"
     CDS             complement(2540..3202)
                     /note="replication protein for the broad host range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /codon_start=1
                     /transl_table=11
                     /product="pBBR1 Rep"
                     /protein_id="WWZ17857.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL
                     HHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAY
                     VVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGP
                     GEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     rep_origin      3203..3974
                     /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 aattaattta tttgacatgc gtgatgttta
      181 gaattataat ttggggacct aggccgcggc cgcgcgaatt cgagctcggt ctcatcacta
      241 gcttatagaa ggagatatac atatgatcat gggaattcat aaaggtgaag aactgttcac
      301 cggtgttgtt ccgatcctgg ttgaactgga tggtgatgtt aacggccaca aattctctgt
      361 tcgtggtgaa ggtgaaggtg atgcaaccaa cggtaaactg accctgaaat tcatctgcac
      421 taccggtaaa ctgccggttc catggccgac tctggtgact accctgacct atggtgttca
      481 gtgtttttct cgttacccgg atcacatgaa gcagcatgat ttcttcaaat ctgcaatgcc
      541 ggaaggttat gtacaggagc gcaccatttc tttcaaagac gatggcacct acaaaacccg
      601 tgcagaggtt aaatttgaag gtgatactct ggtgaaccgt attgaactga aaggcattga
      661 tttcaaagag gacggcaaca tcctgggcca caaactggaa tataacttca actcccataa
      721 cgtttacatc accgcagaca aacagaagaa cggtatcaaa gctaacttca aaattcgcca
      781 taacgttgaa gacggtagcg tacagctggc ggaccactac cagcagaaca ctccgatcgg
      841 tgatggtccg gttctgctgc cggataacca ctacctgtcc acccagtcta aactgtccaa
      901 agacccgaac gaaaagcgcg accacatggt gctgctggag ttcgttactg cagcaggtat
      961 cacgcacggc atggatgaac tctacaaata agggtctctg ttttagagct agaaatagca
     1021 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgctataa
     1081 aacgaaaggc ccagtctttc gactgagcct ttcgttttat aactaggtac ccggggatcc
     1141 tctagagtcg acctgcaggc atgcaagctt gcggccgcgt cgtgactggg aaaaccctgg
     1201 cgactagtct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     1261 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca ggggtcccca
     1321 ataattacga tttaaatttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     1381 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     1441 gagccatatt cagcgtgaaa cgagctgtag ccgtccgcgt ctgaacagca acatggatgc
     1501 ggatctgtat ggctataaat gggcgcgtga taacgtgggt cagagcggcg cgaccattta
     1561 tcgtctgtat ggcaaaccgg atgcgccgga actgtttctg aaacatggca aaggcagcgt
     1621 ggcgaacgat gtgaccgatg aaatggtgcg tctgaactgg ctgaccgaat ttatgccgct
     1681 gccgaccatt aaacatttta ttcgcacccc ggatgatgcg tggctgctga ccaccgcgat
     1741 tccgggcaaa accgcgtttc aggtgctgga agaatatccg gatagcggcg aaaacattgt
     1801 ggatgcgctg gccgtgtttc tgcgtcgtct gcatagcatt ccggtgtgca actgcccgtt
     1861 taacagcgat cgtgtgtttc gtctggccca ggcgcagagc cgtatgaaca acggcctggt
     1921 ggatgcgagc gattttgatg atgaacgtaa cggctggccg gtggaacagg tgtggaaaga
     1981 aatgcataaa ctgctgccgt ttagcccgga tagcgtggtg acccacggcg attttagcct
     2041 ggataacctg attttcgatg aaggcaaact gattggctgc attgatgtgg gccgtgtggg
     2101 cattgcggat cgttatcagg atctggccat tctgtggaac tgcctgggcg aatttagccc
     2161 gagcctgcaa aaacgtctgt ttcagaaata tggcattgat aatccggata tgaacaaact
     2221 gcaatttcat ctgatgctgg atgaattttt ctaataatta attggaccgc ggtccgcgcg
     2281 ttgtcctttt ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat
     2341 ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg
     2401 tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt
     2461 tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga
     2521 ggctggccgt aggccggccc taccggcgcg gcagcgttac ccgtgtcggc ggctccaacg
     2581 gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc
     2641 gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg
     2701 gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg
     2761 tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa
     2821 ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca
     2881 cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga
     2941 tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga
     3001 tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca
     3061 cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct
     3121 catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga
     3181 tttctctgga ctgcgtggcc atgcttatct ccatgcggta ggggtgccgc acggttgcgg
     3241 caccatgcgc aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa
     3301 agtggcagtc aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg
     3361 caatgagctg ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc
     3421 gccctatatc tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc
     3481 ttcggcgcgg ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg
     3541 cggccttcgg ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt
     3601 gaggctcggg gggcaggcgg gcgggcttcg cccttcgact gcccccactc gcataggctt
     3661 gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc
     3721 gccttccact tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt
     3781 ccccagagaa aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg
     3841 tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc
     3901 aggcgcagcc tgtccatcag cttgtccagc agggttgtcc acgggccgag cgaagcgagc
     3961 cagccggtgg ccgctcgcgg ccatcgtcca catatccacg ggctggcaag ggagcgcagc
     4021 gaccgcgcag ggc
//

