LOCUS       PP457288                4045 bp    DNA     circular SYN 16-MAR-2024
DEFINITION  Cloning vector pSpyA-GFP, complete sequence.
ACCESSION   PP457288
VERSION     PP457288.1
KEYWORDS    .
SOURCE      Cloning vector pSpyA-GFP
  ORGANISM  Cloning vector pSpyA-GFP
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4045)
  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  JOURNAL   Unpublished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 4045)
  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..4045
                     /organism="Cloning vector pSpyA-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:3129130"
     regulatory      56..142
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      160..207
                     /regulatory_class="promoter"
                     /note="EM7 synthetic bacterial promoter; EM7"
     gene            275..1003
                     /gene="msfgfp"
     CDS             275..1003
                     /gene="msfgfp"
                     /note="monomeric superfolder green fluorescent protein"
                     /codon_start=1
                     /transl_table=11
                     /product="msfGFP"
                     /protein_id="WWZ17852.1"
                     /translation="MIMGIHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNG
                     KLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI
                     SFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADK
                     QKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYK"
     misc_RNA        1012..1087
                     /note="tracrRNA scaffold for sgRNA; gRNA scaffold"
     regulatory      1089..1132
                     /regulatory_class="terminator"
                     /note="transcription terminator T1 from the E coli rrnB
                     gene; T1"
     regulatory      1225..1319
                     /regulatory_class="terminator"
                     /note="transcription terminator t0 from phage lambda; t0"
     CDS             1451..2266
                     /note="confers resistance to kanamycin in bacteria; KmR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="WWZ17853.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2433..2541
                     /note="incP origin of transfer"
     CDS             complement(2552..3214)
                     /note="replication protein for the broad host range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /codon_start=1
                     /transl_table=11
                     /product="pBBR1 Rep"
                     /protein_id="WWZ17854.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL
                     HHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAY
                     VVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGP
                     GEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     rep_origin      3215..3986
                     /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 aattaacttg ttgacaatta atcatcggca
      181 tagtatatcg gcatagtata atacgacacc ctaggccgcg gccgcgcgaa ttcgagctcg
      241 gtctcatcac tagcttatag aaggagatat acatatgatc atgggaattc ataaaggtga
      301 agaactgttc accggtgttg ttccgatcct ggttgaactg gatggtgatg ttaacggcca
      361 caaattctct gttcgtggtg aaggtgaagg tgatgcaacc aacggtaaac tgaccctgaa
      421 attcatctgc actaccggta aactgccggt tccatggccg actctggtga ctaccctgac
      481 ctatggtgtt cagtgttttt ctcgttaccc ggatcacatg aagcagcatg atttcttcaa
      541 atctgcaatg ccggaaggtt atgtacagga gcgcaccatt tctttcaaag acgatggcac
      601 ctacaaaacc cgtgcagagg ttaaatttga aggtgatact ctggtgaacc gtattgaact
      661 gaaaggcatt gatttcaaag aggacggcaa catcctgggc cacaaactgg aatataactt
      721 caactcccat aacgtttaca tcaccgcaga caaacagaag aacggtatca aagctaactt
      781 caaaattcgc cataacgttg aagacggtag cgtacagctg gcggaccact accagcagaa
      841 cactccgatc ggtgatggtc cggttctgct gccggataac cactacctgt ccacccagtc
      901 taaactgtcc aaagacccga acgaaaagcg cgaccacatg gtgctgctgg agttcgttac
      961 tgcagcaggt atcacgcacg gcatggatga actctacaaa taagggtctc tgttttagag
     1021 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     1081 tcggtgctat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt ataactaggt
     1141 acccggggat cctctagagt cgacctgcag gcatgcaagc ttgcggccgc gtcgtgactg
     1201 ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg acctcagaac
     1261 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     1321 caggggtccc caataattac gatttaaatt tgtgtctcaa aatctctgat gttacattgc
     1381 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1441 aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc gtctgaacag
     1501 caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg gtcagagcgg
     1561 cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg
     1621 caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact ggctgaccga
     1681 atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg cgtggctgct
     1741 gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg
     1801 cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg
     1861 caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa
     1921 caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc cggtggaaca
     1981 ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg tgacccacgg
     2041 cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct gcattgatgt
     2101 gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga actgcctggg
     2161 cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg ataatccgga
     2221 tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat taattggacc
     2281 gcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     2341 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     2401 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     2461 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     2521 cctgctctgc gaggctggcc gtaggccggc cctaccggcg cggcagcgtt acccgtgtcg
     2581 gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct
     2641 gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc
     2701 ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc
     2761 ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt
     2821 cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct
     2881 ggccccacgc cacgcggtca ttgaccacgt aggccgacac ggtgccgggg ccgttgagct
     2941 tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca
     3001 aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc
     3061 ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc
     3121 cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg
     3181 cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc
     3241 gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga caacaattat
     3301 gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa tgagctattg
     3361 cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg atttttaagt
     3421 ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac ccctgcgggg
     3481 ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc cggggcttgt
     3541 tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg gaacccccgc
     3601 actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga ctgcccccac
     3661 tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc
     3721 gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg
     3781 ccggaggctt ttccccagag aaaattaaaa aaattgatgg ggcaaggccg caggccgcgc
     3841 agttggagcc ggtgggtatg tggtcgaagg ctgggtagcc ggtgggcaat ccctgtggtc
     3901 aagctcgtgg gcaggcgcag cctgtccatc agcttgtcca gcagggttgt ccacgggccg
     3961 agcgaagcga gccagccggt ggccgctcgc ggccatcgtc cacatatcca cgggctggca
     4021 agggagcgca gcgaccgcgc agggc
//

