LOCUS       pFH72                   2544 bp    DNA     circular     30-MAY-2019
DEFINITION  Subcloning of sgRNA protospacer/ tRNA-gRNA system with classic gRNA
            backbone/ Position 1.
ACCESSION   urn.local...26-aj37n96
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    66..71
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     overhang        73..76
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="GATT overhang to pFH34"
     Start_codon     77
                     /created_by="hahnf"
                     /label="Transcription Start Signal (G)"
     tRNA            78..154
                     /created_by="hahnf"
                     /Color="#ff7400"
                     /modified_by="hahnf"
                     /label="tRNA Glycine"
     overhang        151..154
                     /created_by="koroleva"
                     /label="TGCA"
     misc_feature    complement(157..162)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             163..740
                     /created_by="koroleva"
                     /label="LacZ"
     misc_feature    741..746
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     gRNA_Scaffold   749..778
                     /created_by="hahnf"
                     /label="classic sgRNA backbone (part)"
     overhang        749..752
                     /created_by="koroleva"
                     /label="GTTT"
     overhang        775..778
                     /created_by="koroleva"
                     /label="AAAT"
     misc_feature    complement(780..785)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(841..1455)
                     /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(1622..2431)
                     /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     2502..2525
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="20849290118205cfb63c94-7f10-4043-946f-4
                     d4bdc12e1e7"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tcgatggtct cagattgaac aaagcaccag tggtctagtg gtagaatagt accctgccac
      121 ggtacagacc cgggttcgat tcccggctgg tgcattgtct tcgcagctgg cacgacaggt
      181 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      241 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      301 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc
      361 aggtcgactc tagaggatcc ccgggtaccg agctcgaatt cactggccgt cgttttacaa
      421 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
      481 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      541 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
      601 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
      661 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
      721 gcttacagac aagctgtgac gaagacaagt tttagagcta gaaatagcaa gttaaaattg
      781 agaccatcat tgtcacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      841 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      901 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      961 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1021 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1081 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1141 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1201 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1261 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1321 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1381 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1441 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1501 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1561 aaaatgaagt tttaaatcaa gcccaatctg aataatgtta caaccaatta accaattctg
     1621 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     1681 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     1741 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     1801 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1861 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1921 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     1981 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     2041 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     2101 attcttctaa tacctggaat gctgtttttc cggggatcgc agtggtgagt aaccatgcat
     2161 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     2221 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     2281 acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga
     2341 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     2401 gcctcgacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     2461 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     2521 tttgagacac gggccagagc tgca
//
