LOCUS       pAK006                  2630 bp    DNA     circular     30-MAY-2019
DEFINITION  Subcloning of sgRNA protospacer/ tRNA-gRNA system with improved
            gRNA backbone/ Position 6.
ACCESSION   urn.local...n-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..91
                     /created_by="hahnf"
                     /label="improved sgRNA backbone (part)"
     overhang        73..76
                     /created_by="koroleva"
                     /label="AGTG"
     tRNA            92..168
                     /created_by="hahnf"
                     /label="tRNA Glycine"
     overhang        165..168
                     /created_by="koroleva"
                     /label="TGCA"
     misc_feature    complement(171..176)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             177..754
                     /created_by="koroleva"
                     /label="LacZ"
     misc_feature    755..760
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     gRNA_Scaffold   763..848
                     /created_by="hahnf"
                     /label="improved sgRNA backbone"
     overhang        763..766
                     /created_by="koroleva"
                     /label="GTTT"
     terminator      849..860
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="Poly-T terminator"
     overhang        861..864
                     /created_by="koroleva"
                     /label="CGCT"
     misc_feature    complement(866..871)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(927..1541)
                     /vntifkey=21
                     /X-Transferred_From="pAK-EL-01; pAK-EL-02; pAK-EL-03;
                     pAK-EL-04; pAK-EL-05"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="pAK-EL-01, pAK-EL-02, pAK-EL-03,
                     pAK-EL-04, pAK-EL-05, pAK007 in pUC57-Kan-mini, pAK008 in
                     pUC57-Kan-mini, pAK009 in pUC57-Kan-mini, pAK010 in
                     pUC57-Kan-mini, pAK011 in pUC57-Kan-mini, pAK012 in
                     pUC57-Kan-mini"
                     /Transferred_Similarity="100.00%"
                     /label="rep(pMB1)"
     CDS             complement(1708..2517)
                     /vntifkey=21
                     /X-Transferred_From="pAK-EL-01; pAK-EL-02; pAK-EL-03;
                     pAK-EL-04; pAK-EL-05"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="pAK-EL-01, pAK-EL-02, pAK-EL-03,
                     pAK-EL-04, pAK-EL-05, pAK007 in pUC57-Kan-mini, pAK008 in
                     pUC57-Kan-mini, pAK009 in pUC57-Kan-mini, pAK010 in
                     pUC57-Kan-mini, pAK011 in pUC57-Kan-mini, pAK012 in
                     pUC57-Kan-mini"
                     /Transferred_Similarity="100.00%"
                     /label="KanR"
     primer_bind     2588..2611
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="1295679172649259b55bee-4166-4333-8e04-f
                     bdbde95cf12"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tcgatggtct caagtggcac cgagtcggtg caacaaagca ccagtggtct agtggtagaa
      121 tagtaccctg ccacggtaca gacccgggtt cgattcccgg ctggtgcatt gtcttcgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg aattcactgg
      421 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      481 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      541 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
      601 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
      661 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      721 tgctcccggc atccgcttac agacaagctg tgacgaagac aagtttcaga gctatgctgg
      781 aaacagcata gcaagttgaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
      841 gtcggtgctt tttttttttt cgcttgagac catcattgtc acatgtgagc aaaaggccag
      901 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      961 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1021 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1081 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1141 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1201 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1261 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1321 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1381 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1441 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1501 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1561 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1621 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaagccc aatctgaata
     1681 atgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     1741 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1801 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1861 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1921 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     1981 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     2041 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     2101 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     2161 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg
     2221 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     2281 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     2341 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg
     2401 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     2461 agcatccatg ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac
     2521 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     2581 atcttgtgca atgtaacatc agagattttg agacacgggc cagagctgca
//
