LOCUS      pSEVA-gRic6T                4521 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  E.coli-Pseudomonas putida KT2440 shuttle plasmid containing sgRNA 
            targeting the KT2440 genome, and homologous arms..
ACCESSION   .
VERSION     .
KEYWORDS    pSEVA-gRic6T
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4521)
  AUTHORS   Sun J, Wang Q, Jiang Y, Wen Z, Yang L, Wu J, Yang S
  TITLE     Genome editing and transcriptional repression in Pseudomonas putida 
            KT2440 via the type II CRISPR system.
  JOURNAL   Microb Cell Fact. 2018 Mar 13;17(1):41. doi: 
            10.1186/s12934-018-0887-x.
  PUBMED    29534717
REFERENCE   2  (bases 1 to 4521)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..4521
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        25..59
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        80..155
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      233..242
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     terminator      1247..1341
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1372..1400
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            2314..2422
                     /note="incP origin of transfer"
     rep_origin      2516..3104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3005..3024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3158..3991)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      4005..4356
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      4429..4515
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaatt gacaccatga attcttgaca gctagctcag tcctaggtat aatgctagca
       61 ggcagaaaat cgcaatcgtg ttttagagct agaaatagca agttaaaata aggctagtcc
      121 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaggagct cggtacccgg
      181 ggatccggca gctctaccgc atacaccttg gccttcaaac gctgggcggc ctccatggcg
      241 gcgttgaccg tgcgcgggtc ggagtagtgg ctcttgagca cggccacgct ctgcgtttcg
      301 tccactcgag acagcttcag tacgtgctcg aacatctggg tcagttgtgc attgctcacc
      361 ggcatcgggg ttctcctggg tggcgttgac aggggcgcac catggcagtg cctggtgctg
      421 catctcggtg caaaataata gtgtgtacac cctacttgtg cagatgaaaa cgcatcctgc
      481 cagcgagcgc aagtgatctg ttcaaacgtt acctatccac acattgacca gtatttacgg
      541 taacaatcaa ccggatttcg tttcgatcag ctgtttgaaa aatattttca gctggctctt
      601 ttcaaaggtc atttgtagcg tatacgctct aaatcgtcag gcggttgagc cgctggccac
      661 ctatcagaac aagaggagac atacccgcac cttcgtcgct ggtggtaacg tcagcgccaa
      721 tggcatccgc cagcattacc tgcgctatgg cggcaagggc catgcgctga tcctggtgcc
      781 gggcatcacc agcccagcga tcacctgggg ctttgtcgcc gagcgccttg ggcattattt
      841 cgacacctat gtgctggatg tgcgcgggcg tggcttgtct tcaagcggcc ccgacctgga
      901 ctacggcacc gacgcttgtg ctgcggatat tcccgccttt gccgccgcgc tgggcctgga
      961 tagctaccat ctgctcggtc actcgatggg ggcgcgtttt gccatccgcg cagccgccca
     1021 aggggcaccg ggactgcagc ggctggtgct ggtcgaccca ccggtgtccg ggccgggccg
     1081 ccgcgcctac ccgagcaagt tgccgtggta cgtcgattcg atccgccaag ccacggttgg
     1141 catgagcggc gatgacatgc gcgccttctg tgccacctgg agcgacctgc aggcatgcaa
     1201 gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata
     1261 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     1321 gcgtttttta ttggtgagaa tccaggggtc cccaataatt acgatttaaa tttgacataa
     1381 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     1441 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     1501 tgtttttttg tacagcctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1561 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     1621 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc
     1681 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac
     1741 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc
     1801 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc
     1861 aagtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggagagcac
     1921 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt
     1981 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat
     2041 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc
     2101 taacaattcg ttcaagccga gatcgacttc ccggccgcgg agttgttcgg taaattggac
     2161 aacggtccgc gcgttgtcct tttccgctgc ataaccctgc ttcggggtca ttatagcgat
     2221 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
     2281 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
     2341 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
     2401 tcctgctctg cgaggctggc cgtaggccgg ccgataatct catgaccaaa atcccttaac
     2461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2641 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2881 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2941 aggcggacag gcatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3121 ccgtgaaagg caggccggtc cgtggtggcc acggcctcta ggccagatcc agcggcatct
     3181 gggttagtcg agcgcgggcc gcttcccatg tctcaccagg gcgagcctgt ttcgcgatct
     3241 cagcatctga aatcttcccg gccttgcgct tcgctggggc cttacccacc gccttggcgg
     3301 gcttcttcgg tccaaaactg aacaacagat gtgtgacctt gcgcccggtc tttcgctgcg
     3361 cccactccac ctgtagcggg ctgtgctcgt tgatctgcgt cacggctgga tcaagcactc
     3421 gcaacttgaa gtccttgatc gagggatacc ggccttccag ttgaaaccac tttcgcagct
     3481 ggtcaatttc tatttcgcgc tggccgatgc tgtcccattg catgagcagc tcgtaaagcc
     3541 tgatcgcgtg ggtgctgtcc atcttggcca cgtcagccaa ggcgtatttg gtgaactgtt
     3601 tggtgagttc cgtcaggtac ggcagcatgt ctttggtgaa cctgagttct acacggccct
     3661 caccctcccg gtagatgatt gtttgcaccc agccggtaat catcacactc ggtcttttcc
     3721 ccttgccatt gggctcttgg gttaaccgga cttcccgccg tttcaggcgc agggccgctt
     3781 ctttgagctg gttgtaggaa gattcgatag ggacacccgc catcgtcgct atgtcctccg
     3841 ccgtcactga atacatcact tcatcggtga caggctcgct cctcttcacc tggctaatac
     3901 aggccagaac gatccgctgt tcctgaacac tgaggcgata cgcggcctcg accagggcat
     3961 tgcttttgta aaccattggg ggtgaggcca cgttcgacat tccttgtgta taaggggaca
     4021 ctgtatctgc gtcccacaat acaacaaatc cgtcccttta caacaacaaa tccgtccctt
     4081 cttaacaaca aatccgtccc ttaatggcaa caaatccgtc cctttttaaa ctctacaggc
     4141 cacggattac gtggcctgta gacgtcctaa aaggtttaaa agggaaaagg aagaaaaggg
     4201 tggaaacgca aaaaacgcac cactacgtgg ccccgttggg gccgcatttg tgcccctgaa
     4261 ggggcggggg aggcgtctgg gcaatccccg ttttaccagt cccctatcgc cgcctgagag
     4321 ggcgcaggaa gcgagtaatc agggtatcga ggcggattca cccttggcgt ccaaccagcg
     4381 gcaccagcgg cgcctgagag gggcgcgccc agctgtctag ggcggcggat ttgtcctact
     4441 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
     4501 ctttcgtttt atttgatgcc t
//