LOCUS       PP457281                4240 bp    DNA     circular SYN 16-MAR-2024
DEFINITION  Cloning vector pMRS-BG35-GFP, complete sequence.
ACCESSION   PP457281
VERSION     PP457281.1
KEYWORDS    .
SOURCE      Cloning vector pMRS-BG35-GFP
  ORGANISM  Cloning vector pMRS-BG35-GFP
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4240)
  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  JOURNAL   Unpublished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 4240)
  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..4240
                     /organism="Cloning vector pMRS-BG35-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:3129119"
     regulatory      51..91
                     /regulatory_class="promoter"
                     /note="BG35 constitutive promoter (Zobel et al., 2015);
                     BG35"
     misc_feature    98..174
                     /note="BCD2 bicistronic linker (Zobel et al., 2015); BCD2"
     regulatory      166..174
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
     gene            181..909
                     /gene="msfgfp"
     CDS             181..909
                     /gene="msfgfp"
                     /note="monomeric superfolder green fluorescent protein"
                     /codon_start=1
                     /transl_table=11
                     /product="msfGFP"
                     /protein_id="WWZ17836.1"
                     /translation="MIMGIHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNG
                     KLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI
                     SFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADK
                     QKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYK"
     regulatory      1009..1103
                     /regulatory_class="terminator"
                     /note="transcription terminator t0 from phage lambda; t0"
     misc_feature    1108..1306
                     /note="Tn7R"
     CDS             1416..2231
                     /note="confers resistance to kanamycin in bacteria; KmR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="WWZ17837.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2398..2506
                     /note="incP origin of transfer"
     rep_origin      2528..2916
                     /note="replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication;
                     R6K"
     misc_feature    2925..3090
                     /note="Tn7L"
     regulatory      3104..3190
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     protein_bind    3225..3272
                     /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(3402..3935)
                     /gene="aacC1"
     CDS             complement(3402..3935)
                     /gene="aacC1"
                     /note="confers resistance to gentamycin; GmR"
                     /codon_start=1
                     /transl_table=11
                     /product="gentamycin acetyltransferase"
                     /protein_id="WWZ17838.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAAL
                     DLFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQP
                     RSEIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKL
                     GIREEVMHFDIDPSTAT"
     regulatory      complement(4124..4152)
                     /regulatory_class="promoter"
                     /note="intI1 (promoter lies within the coding sequence);
                     class 1 integron promoter; Pc promoter"
     protein_bind    4193..4240
                     /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 tttatttgac
       61 atgcgtgatg tttagaatta taatttgggg acctagggcc caagttcact taaaaaggag
      121 atcaacaatg aaagcaattt tcgtactgaa acatcttaat catgctaagg aggttttcta
      181 atgatcatgg gaattcataa aggtgaagaa ctgttcaccg gtgttgttcc gatcctggtt
      241 gaactggatg gtgatgttaa cggccacaaa ttctctgttc gtggtgaagg tgaaggtgat
      301 gcaaccaacg gtaaactgac cctgaaattc atctgcacta ccggtaaact gccggttcca
      361 tggccgactc tggtgactac cctgacctat ggtgttcagt gtttttctcg ttacccggat
      421 cacatgaagc agcatgattt cttcaaatct gcaatgccgg aaggttatgt acaggagcgc
      481 accatttctt tcaaagacga tggcacctac aaaacccgtg cagaggttaa atttgaaggt
      541 gatactctgg tgaaccgtat tgaactgaaa ggcattgatt tcaaagagga cggcaacatc
      601 ctgggccaca aactggaata taacttcaac tcccataacg tttacatcac cgcagacaaa
      661 cagaagaacg gtatcaaagc taacttcaaa attcgccata acgttgaaga cggtagcgta
      721 cagctggcgg accactacca gcagaacact ccgatcggtg atggtccggt tctgctgccg
      781 gataaccact acctgtccac ccagtctaaa ctgtccaaag acccgaacga aaagcgcgac
      841 cacatggtgc tgctggagtt cgttactgca gcaggtatca cgcacggcat ggatgaactc
      901 tacaaataag aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
      961 aagcttgcgg ccgcgtcgtg actgggaaaa ccctggcgac tagtcttgga ctcctgttga
     1021 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     1081 gggcgttttt tattggtgag aatccagcca cgcccctctt taatacgacg ggcaatttgc
     1141 acttcagaaa atgaagagtt tgctttagcc ataacaaaag tccagtatgc tttttcacag
     1201 cataactgga ctgatttcag tttacaacta ttctgtctag tttaagactt tattgtcata
     1261 gtttagatct attttgttca gtttaagact ttattgtccg cccacaattt aaatttgtgt
     1321 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1381 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcag cgtgaaacga
     1441 gctgtagccg tccgcgtctg aacagcaaca tggatgcgga tctgtatggc tataaatggg
     1501 cgcgtgataa cgtgggtcag agcggcgcga ccatttatcg tctgtatggc aaaccggatg
     1561 cgccggaact gtttctgaaa catggcaaag gcagcgtggc gaacgatgtg accgatgaaa
     1621 tggtgcgtct gaactggctg accgaattta tgccgctgcc gaccattaaa cattttattc
     1681 gcaccccgga tgatgcgtgg ctgctgacca ccgcgattcc gggcaaaacc gcgtttcagg
     1741 tgctggaaga atatccggat agcggcgaaa acattgtgga tgcgctggcc gtgtttctgc
     1801 gtcgtctgca tagcattccg gtgtgcaact gcccgtttaa cagcgatcgt gtgtttcgtc
     1861 tggcccaggc gcagagccgt atgaacaacg gcctggtgga tgcgagcgat tttgatgatg
     1921 aacgtaacgg ctggccggtg gaacaggtgt ggaaagaaat gcataaactg ctgccgttta
     1981 gcccggatag cgtggtgacc cacggcgatt ttagcctgga taacctgatt ttcgatgaag
     2041 gcaaactgat tggctgcatt gatgtgggcc gtgtgggcat tgcggatcgt tatcaggatc
     2101 tggccattct gtggaactgc ctgggcgaat ttagcccgag cctgcaaaaa cgtctgtttc
     2161 agaaatatgg cattgataat ccggatatga acaaactgca atttcatctg atgctggatg
     2221 aatttttcta ataattaatt ggaccgcggt ccgcgcgttg tccttttccg ctgcataacc
     2281 ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac
     2341 aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg
     2401 caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc
     2461 gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccggcg
     2521 cgccccatgt cagccgttaa gtgttcctgt gtcactcaaa attgctttga gaggctctaa
     2581 gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaggct
     2641 ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa
     2701 gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag
     2761 agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa
     2821 catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac
     2881 gtactatcaa caggttgaac tgctgatctt cagatcggcg cgcctgtggg cggacaaaat
     2941 agttgggaac tgggaggggt ggaaatggag tttttaagga ttatttaggg aagagtgaca
     3001 aaatagatgg gaactgggtg tagcgtcgta agctaatacg aaaattaaaa atgacaaaat
     3061 agtttggaac tagatttcac ttatctggtt tctagggcgg cggatttgtc ctactcagga
     3121 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     3181 gttttatttg atgccgacgt ccggacgatc gaattgggga tcttgaagta cctattccga
     3241 agttcctatt ctctagaaag tataggaact tcagagcgct tttgaagctg atgtgcttaa
     3301 aaacttactc aatggaattc tagatcgaat tggccgcggc gttgtgacaa tttaccgaac
     3361 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     3421 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     3481 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     3541 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3601 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3661 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3721 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3781 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3841 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3901 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3961 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     4021 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     4081 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     4141 ggcttatgtc aatccgatcc attgctgttg acaaagggaa tcaggggatc ttgaagttcc
     4201 tattccgaag ttcctattct ctagaaagta taggaacttc
//

