LOCUS       pAK010                  2595 bp    DNA     circular     30-MAY-2019
DEFINITION  Subcloning of sgRNA protospacer/ tRNA-gRNA system with classic gRNA
            backbone/ Position 4.
ACCESSION   urn.local...r-aj37jvy
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"
     gRNA_Scaffold   73..102
                     /created_by="hahnf"
                     /label="classic sgRNA backbone (part)"
     overhang        73..76
                     /created_by="koroleva"
                     /label="CAAC"
     tRNA            103..179
                     /created_by="hahnf"
                     /label="tRNA Glycine"
     overhang        176..179
                     /created_by="koroleva"
                     /label="TGCA"
     CDS             188..765
                     /created_by="koroleva"
                     /label="LacZ"
     gRNA_Scaffold   774..829
                     /created_by="hahnf"
                     /label="classic sgRNA backbone (part)"
     overhang        774..777
                     /created_by="koroleva"
                     /label="GTTT"
     overhang        826..829
                     /created_by="koroleva"
                     /label="GAAA"
     misc_feature    complement(831..836)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(892..1506)
                     /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(1673..2482)
                     /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     2553..2576
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="12969662732637b5989c7c-aa64-4b90-9d25-8
                     293f3549894"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tcgatggtct cacaacttga aaaagtggca ccgagtcggt gcaacaaagc accagtggtc
      121 tagtggtaga atagtaccct gccacggtac agacccgggt tcgattcccg gctggtgcat
      181 tgtcttcgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      241 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      301 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      361 gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc
      421 gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      481 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      541 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
      601 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
      661 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
      721 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacgaaga caagttttag
      781 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaat gagaccatca
      841 ttgtcacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1321 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1621 ttttaaatca agcccaatct gaataatgtt acaaccaatt aaccaattct gattagaaaa
     1681 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1741 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1801 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1861 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1921 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1981 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     2041 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     2101 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     2161 atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2221 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2281 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2341 gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg acattatcgc
     2401 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgacg
     2461 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     2521 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     2581 cgggccagag ctgca
//
