LOCUS       pFH103                  2813 bp    DNA     circular UNA 18-NOV-2019
DEFINITION  Subcloning of protospacer into LbCas12a crRNA backbone, overhang to
            pFH35 AtU6-26p.
ACCESSION   urn.local...1v-ajo6si0
VERSION     urn.local...1v-ajo6si0
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    2..7
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     overhang        9..12
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="ATTG to AtU6-26p pFH35"
     gRNA_Scaffold   13..33
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="LbCas12a crRNA backbone"
     overhang        30..33
                     /created_by="hahnf"
                     /label="AGAT"
     misc_feature    complement(36..41)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             42..619
                     /created_by="hahnf"
                     /label="lacZ"
     misc_feature    620..625
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     Stop_codon      628..636
                     /created_by="hahnf"
                     /label="STOP"
     overhang        628..631
                     /created_by="hahnf"
                     /label="TTTT"
     overhang        637..640
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="CGCT overhang to Level1 backbone"
     misc_feature    complement(642..647)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(878..1492)
                     /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(1617..2432)
                     /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="98.90%"
                     /label="KanR"
     primer_bind     2503..2526
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="24556894963731b063b2e7-c364-41aa-8d5a-6
                     54813c50194"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 gggtctcaat tgtaatttct actaagtgta gatttgtctt cgcagctggc acgacaggtt
       61 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      121 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      181 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
      241 ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
      301 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      361 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      421 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      481 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      541 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      601 cttacagaca agctgtgacg aagacaattt ttttttcgct tgagaccgag gctaggtgga
      661 ggctcagtga tgataagtct gcgatggtgg atgcatgtgt catggtcata gctgtttcct
      721 gtgtgaaatt gttatccgct cagagggcac aatcctattc cgcgctatcc gacaatctcc
      781 aagacattag gtggagttca gttcggcgta tggcatatgt cgctggaaag aacatgtgag
      841 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      901 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      961 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1021 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1081 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1141 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1201 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1261 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1321 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1381 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1441 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1501 ctacggggtc tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag cgtaaatggg
     1561 tagggggctt caaatcgtcc gctctgccag tgttacaacc aattaacaaa ttctgattag
     1621 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     1681 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     1741 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     1801 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     1861 tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca
     1921 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     1981 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     2041 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     2101 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     2161 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     2221 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     2281 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     2341 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     2401 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     2461 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     2521 ttttgagaca caacgtggct ttcccccgcc gctctagaac tagtggatcc aaataaaacg
     2581 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgc attatacgag
     2641 acgtccaggt tgggatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
     2701 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     2761 tcatgcataa tccgcacgca tctggaataa ggaagtgcca ttccgcctga cct
//
