LOCUS       pFH104                  2812 bp    DNA     circular UNA 18-NOV-2019
DEFINITION  Subcloning of protospacer into FnCas12a crRNA backbone, overhang to
            pFH35 AtU6-26p.
ACCESSION   urn.local...1w-ajo6si0
VERSION     urn.local...1w-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..32
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="FnCas12a crRNA backbone"
     overhang        29..32
                     /created_by="hahnf"
                     /label="AGAT"
     misc_feature    complement(35..40)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             41..618
                     /created_by="hahnf"
                     /label="lacZ"
     misc_feature    619..624
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     Stop_codon      627..635
                     /created_by="hahnf"
                     /label="STOP"
     overhang        627..630
                     /created_by="hahnf"
                     /label="TTTT"
     overhang        636..639
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="CGCT overhang to Level1 backbone"
     misc_feature    complement(641..646)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(877..1491)
                     /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(1616..2431)
                     /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     2502..2525
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="24557275296831d9f69955-a893-4e5c-ab3e-b
                     eab176b4711"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 gggtctcaat tgtaatttct actgttgtag atttgtcttc gcagctggca cgacaggttt
       61 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      121 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      181 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
      241 gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg
      301 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      361 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      421 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      481 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      541 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      601 ttacagacaa gctgtgacga agacaatttt tttttcgctt gagaccgagg ctaggtggag
      661 gctcagtgat gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg
      721 tgtgaaattg ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca
      781 agacattagg tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc
      841 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      901 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      961 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1021 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1081 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1141 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1201 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1261 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1321 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1381 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1441 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1501 tacggggtct gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt
     1561 agggggcttc aaatcgtccg ctctgccagt gttacaacca attaacaaat tctgattaga
     1621 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     1681 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     1741 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     1801 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     1861 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     1921 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     1981 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     2041 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     2101 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     2161 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     2221 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     2281 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     2341 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     2401 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     2461 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     2521 tttgagacac aacgtggctt tcccccgccg ctctagaact agtggatcca aataaaacga
     2581 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga
     2641 cgtccaggtt gggatacctg aaacaaaacc catcgtacgg ccaaggaagt ctccaataac
     2701 tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     2761 catgcataat ccgcacgcat ctggaataag gaagtgccat tccgcctgac ct
//
