LOCUS       pAK005                  2604 bp    DNA     circular     30-MAY-2019
DEFINITION  Subcloning of sgRNA protospacer/ tRNA-gRNA system with improved
            gRNA backbone/ Position 5.
ACCESSION   urn.local...m-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..96
                     /created_by="hahnf"
                     /label="improved sgRNA backbone (part)"
     overhang        73..76
                     /created_by="koroleva"
                     /label="GAAA"
     tRNA            97..173
                     /created_by="hahnf"
                     /label="tRNA Glycine"
     overhang        170..173
                     /created_by="koroleva"
                     /label="TGCA"
     misc_feature    complement(176..181)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             182..759
                     /created_by="koroleva"
                     /label="LacZ"
     misc_feature    760..765
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     gRNA_Scaffold   768..838
                     /created_by="hahnf"
                     /label="improved sgRNA backbone (part)"
     overhang        768..771
                     /created_by="koroleva"
                     /label="GTTT"
     overhang        835..838
                     /created_by="koroleva"
                     /label="AGTG"
     misc_feature    complement(840..845)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(901..1515)
                     /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(1682..2491)
                     /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     2562..2585
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="1295624007652761dad34d-a9ce-4a67-87ed-9
                     e0cd62a53a8"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tcgatggtct cagaaaaagt ggcaccgagt cggtgcaaca aagcaccagt ggtctagtgg
      121 tagaatagta ccctgccacg gtacagaccc gggttcgatt cccggctggt gcattgtctt
      181 cgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      241 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      301 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      361 gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc
      421 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      481 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      541 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
      601 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
      661 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      721 ttgtctgctc ccggcatccg cttacagaca agctgtgacg aagacaagtt tcagagctat
      781 gctggaaaca gcatagcaag ttgaaataag gctagtccgt tatcaacttg aaaaagtgtg
      841 agaccatcat tgtcacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      901 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      961 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1021 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1081 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1141 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1201 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1261 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1321 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1381 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1441 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1501 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1561 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1621 aaaatgaagt tttaaatcaa gcccaatctg aataatgtta caaccaatta accaattctg
     1681 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     1741 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     1801 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     1861 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1921 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1981 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     2041 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     2101 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     2161 attcttctaa tacctggaat gctgtttttc cggggatcgc agtggtgagt aaccatgcat
     2221 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     2281 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     2341 acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga
     2401 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     2461 gcctcgacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     2521 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     2581 tttgagacac gggccagagc tgca
//
