LOCUS       pFH49                   2552 bp    DNA     circular UNA 30-MAY-2019
DEFINITION  Subcloning of sgRNA protospacer/ tRNA-gRNA system with improved
            gRNA backbone/ Position 1.
ACCESSION   urn.local...22-aj37n96
VERSION     urn.local...22-aj37n96
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    64..69
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     overhang        71..74
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="GATT overhang to pFH34"
     Start_codon     75
                     /created_by="hahnf"
                     /label="Transcription Start Signal (G)"
     tRNA            76..152
                     /created_by="hahnf"
                     /Color="#ff7400"
                     /Transferred_From="pFH26, pFH27, pFH28"
                     /Transferred_Similarity="100.00%"
                     /label="tRNA Glycine"
     overhang        149..152
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="TGCA"
     misc_feature    complement(155..160)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             161..738
                     /created_by="koroleva"
                     /Transferred_From="AK001 in pUC57-Kan-mini, AK002 in
                     pUC57-Kan-mini, pAK001 in pUC57-Kan-mini, pAK002 in
                     pUC57-Kan-mini"
                     /Transferred_Similarity="100.00%"
                     /label="LacZ"
     misc_feature    739..744
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     overhang        747..750
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="GTTT"
     overhang        783..786
                     /created_by="hahnf"
                     /label="AAAT"
     misc_feature    complement(788..793)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(849..1463)
                     /vntifkey=21
                     /Transferred_From="pAK-EL-01, pAK-EL-02, pAK-EL-03,
                     pAK-EL-04, pAK-EL-05"
                     /Transferred_Similarity="100.00%"
                     /label="rep(pMB1)"
     CDS             complement(1630..2439)
                     /vntifkey=21
                     /Transferred_From="pAK-EL-01, pAK-EL-02, pAK-EL-03,
                     pAK-EL-04, pAK-EL-05"
                     /Transferred_Similarity="100.00%"
                     /label="KanR"
     primer_bind     2510..2533
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="2492998740745d7af307b-ad34-4a87-87a8-2b
                     dffb2c6aba"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /Transferred_From="AK001 in pUC57-Kan-mini"
                     /Transferred_Similarity="100.00%"
                     /label="FH32"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tctggtctca gattgaacaa agcaccagtg gtctagtggt agaatagtac cctgccacgg
      121 tacagacccg ggttcgattc ccggctggtg cattgtcttc gcagctggca cgacaggttt
      181 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      241 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      301 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
      361 gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg
      421 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      481 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      541 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      601 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      661 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      721 ttacagacaa gctgtgacga agacaagttt cagagctatg ctggaaacag catagcaagt
      781 tgaaattgag accatcattg tcacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1561 ttaaattaaa aatgaagttt taaatcaagc ccaatctgaa taatgttaca accaattaac
     1621 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     1681 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     1741 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     1801 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     1861 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     1921 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     1981 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2041 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2101 atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa
     2161 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     2221 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     2281 tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga
     2341 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     2401 taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     2461 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     2521 tcagagattt tgagacacgg gccagagctg ca
//
