LOCUS       PP457277                4228 bp    DNA     circular SYN 16-MAR-2024
DEFINITION  Cloning vector pMRS-GFP, complete sequence.
ACCESSION   PP457277
VERSION     PP457277.1
KEYWORDS    .
SOURCE      Cloning vector pMRS-GFP
  ORGANISM  Cloning vector pMRS-GFP
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4228)
  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  JOURNAL   Unpublished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 4228)
  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..4228
                     /organism="Cloning vector pMRS-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:3129121"
     misc_feature    82..162
                     /note="BCD2 bicistronic linker (Zobel et al., 2015); BCD2"
     regulatory      154..162
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
     gene            169..897
                     /gene="msfgfp"
     CDS             169..897
                     /gene="msfgfp"
                     /note="monomeric superfolder green fluorescent protein"
                     /codon_start=1
                     /transl_table=11
                     /product="msfGFP"
                     /protein_id="WWZ17821.1"
                     /translation="MIMGIHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNG
                     KLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI
                     SFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADK
                     QKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYK"
     regulatory      997..1091
                     /regulatory_class="terminator"
                     /note="transcription terminator t0 from phage lambda; t0"
     misc_feature    1096..1294
                     /note="Tn7R"
     CDS             1404..2219
                     /note="confers resistance to kanamycin in bacteria; KmR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="WWZ17822.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2386..2494
                     /note="incP origin of transfer"
     rep_origin      2515..2904
                     /note="replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication;
                     R6K"
     misc_feature    2913..3078
                     /note="Tn7L"
     regulatory      3092..3178
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     protein_bind    3213..3260
                     /note="from the Saccharomyces cerevisiae 2u plasmid;
                     FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011); FRT"
                     /bound_moiety="FLP recombinase"
     gene            complement(3390..3923)
                     /gene="aacC1"
     CDS             complement(3390..3923)
                     /gene="aacC1"
                     /note="confers resistance to gentamycin; GmR"
                     /codon_start=1
                     /transl_table=11
                     /product="gentamycin acetyltransferase"
                     /protein_id="WWZ17823.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAAL
                     DLFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQP
                     RSEIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKL
                     GIREEVMHFDIDPSTAT"
     regulatory      complement(4112..4140)
                     /regulatory_class="promoter"
                     /note="intI1 (promoter lies within the coding sequence);
                     class 1 integron promoter; Pc promoter"
     protein_bind    4181..4228
                     /note="from the Saccharomyces cerevisiae 2u plasmid;
                     FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011); FRT"
                     /bound_moiety="FLP recombinase"
ORIGIN      
        1 agagcgcttt tgaagctaat tcgatcatgc atgagcgacg tcttaattaa agcggataac
       61 aatttcacac aggaggccgc ctagggccca agttcactta aaaaggagat caacaatgaa
      121 agcaattttc gtactgaaac atcttaatca tgctaaggag gttttctaat gatcatggga
      181 attcataaag gtgaagaact gttcaccggt gttgttccga tcctggttga actggatggt
      241 gatgttaacg gccacaaatt ctctgttcgt ggtgaaggtg aaggtgatgc aaccaacggt
      301 aaactgaccc tgaaattcat ctgcactacc ggtaaactgc cggttccatg gccgactctg
      361 gtgactaccc tgacctatgg tgttcagtgt ttttctcgtt acccggatca catgaagcag
      421 catgatttct tcaaatctgc aatgccggaa ggttatgtac aggagcgcac catttctttc
      481 aaagacgatg gcacctacaa aacccgtgca gaggttaaat ttgaaggtga tactctggtg
      541 aaccgtattg aactgaaagg cattgatttc aaagaggacg gcaacatcct gggccacaaa
      601 ctggaatata acttcaactc ccataacgtt tacatcaccg cagacaaaca gaagaacggt
      661 atcaaagcta acttcaaaat tcgccataac gttgaagacg gtagcgtaca gctggcggac
      721 cactaccagc agaacactcc gatcggtgat ggtccggttc tgctgccgga taaccactac
      781 ctgtccaccc agtctaaact gtccaaagac ccgaacgaaa agcgcgacca catggtgctg
      841 ctggagttcg ttactgcagc aggtatcacg cacggcatgg atgaactcta caaataagaa
      901 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttgcggcc
      961 gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata gatccagtaa
     1021 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1081 ttggtgagaa tccagccacg cccctcttta atacgacggg caatttgcac ttcagaaaat
     1141 gaagagtttg ctttagccat aacaaaagtc cagtatgctt tttcacagca taactggact
     1201 gatttcagtt tacaactatt ctgtctagtt taagacttta ttgtcatagt ttagatctat
     1261 tttgttcagt ttaagacttt attgtccgcc cacaatttaa atttgtgtct caaaatctct
     1321 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1381 taaacagtaa tacaaggggt gttatgagcc atattcagcg tgaaacgagc tgtagccgtc
     1441 cgcgtctgaa cagcaacatg gatgcggatc tgtatggcta taaatgggcg cgtgataacg
     1501 tgggtcagag cggcgcgacc atttatcgtc tgtatggcaa accggatgcg ccggaactgt
     1561 ttctgaaaca tggcaaaggc agcgtggcga acgatgtgac cgatgaaatg gtgcgtctga
     1621 actggctgac cgaatttatg ccgctgccga ccattaaaca ttttattcgc accccggatg
     1681 atgcgtggct gctgaccacc gcgattccgg gcaaaaccgc gtttcaggtg ctggaagaat
     1741 atccggatag cggcgaaaac attgtggatg cgctggccgt gtttctgcgt cgtctgcata
     1801 gcattccggt gtgcaactgc ccgtttaaca gcgatcgtgt gtttcgtctg gcccaggcgc
     1861 agagccgtat gaacaacggc ctggtggatg cgagcgattt tgatgatgaa cgtaacggct
     1921 ggccggtgga acaggtgtgg aaagaaatgc ataaactgct gccgtttagc ccggatagcg
     1981 tggtgaccca cggcgatttt agcctggata acctgatttt cgatgaaggc aaactgattg
     2041 gctgcattga tgtgggccgt gtgggcattg cggatcgtta tcaggatctg gccattctgt
     2101 ggaactgcct gggcgaattt agcccgagcc tgcaaaaacg tctgtttcag aaatatggca
     2161 ttgataatcc ggatatgaac aaactgcaat ttcatctgat gctggatgaa tttttctaat
     2221 aattaattgg accgcggtcc gcgcgttgtc cttttccgct gcataaccct gcttcggggt
     2281 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     2341 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     2401 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     2461 gctcaacggg aatcctgctc tgcgaggctg gccgtaggcc ggccggcgcg ccccatgtca
     2521 gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt
     2581 gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaggcttt aaaagcctta
     2641 tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta
     2701 tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt
     2761 tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta
     2821 gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca
     2881 ggttgaactg ctgatcttca gatcggcgcg cctgtgggcg gacaaaatag ttgggaactg
     2941 ggaggggtgg aaatggagtt tttaaggatt atttagggaa gagtgacaaa atagatggga
     3001 actgggtgta gcgtcgtaag ctaatacgaa aattaaaaat gacaaaatag tttggaacta
     3061 gatttcactt atctggtttc tagggcggcg gatttgtcct actcaggaga gcgttcaccg
     3121 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     3181 gccgacgtcc ggacgatcga attggggatc ttgaagtacc tattccgaag ttcctattct
     3241 ctagaaagta taggaacttc agagcgcttt tgaagctgat gtgcttaaaa acttactcaa
     3301 tggaattcta gatcgaattg gccgcggcgt tgtgacaatt taccgaacaa ctccgcggcc
     3361 gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag
     3421 tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg
     3481 taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga
     3541 ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag
     3601 atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca
     3661 acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc
     3721 ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga
     3781 ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg
     3841 cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta
     3901 acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc
     3961 gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca
     4021 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     4081 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
     4141 tccgatccat tgctgttgac aaagggaatc aggggatctt gaagttccta ttccgaagtt
     4201 cctattctct agaaagtata ggaacttc
//

