LOCUS       pFH87                   2917 bp    DNA     circular UNA 03-JUN-2019
DEFINITION  Subcloning of protospacer into StCas9 sgRNA backbone, overhang to
            pFH33 TaU3p.
ACCESSION   urn.local...1n-ajo6shz
VERSION     urn.local...1n-ajo6shz
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"
                     /label="AGCA to TaU3p pFH33"
     misc_feature    complement(15..20)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             21..598
                     /created_by="hahnf"
                     /label="lacZ"
     misc_feature    599..604
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     gRNA_Scaffold   607..731
                     /created_by="hahnf"
                     /label="StCas9 sgRNA scaffold"
     overhang        607..610
                     /created_by="hahnf"
                     /label="GTTT"
     Stop_codon      732..740
                     /created_by="hahnf"
                     /label="STOP"
     overhang        741..744
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="CGCT overhang to Level1 backbone"
     misc_feature    complement(746..751)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(982..1596)
                     /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(1721..2536)
                     /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     2607..2630
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="24556360840306f5a93fb6-290f-458c-9945-1
                     088b4bea747"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /label="FH32"
ORIGIN      
        1 gggtctcaag cattgtcttc gcagctggca cgacaggttt cccgactgga aagcgggcag
       61 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      121 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      181 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc
      241 gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
      301 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
      361 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
      421 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
      481 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
      541 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgacga
      601 agacaagttt ttgtactctc aagatttaag taactgtaca acgaaactta cacagttact
      661 taaatcttgc agaagctaca aagataaggc ttcatgccga aatcaacacc ctgtcatttt
      721 atggcagggt gttttttttt cgcttgagac cgaggctagg tggaggctca gtgatgataa
      781 gtctgcgatg gtggatgcat gtgtcatggt catagctgtt tcctgtgtga aattgttatc
      841 cgctcagagg gcacaatcct attccgcgct atccgacaat ctccaagaca ttaggtggag
      901 ttcagttcgg cgtatggcat atgtcgctgg aaagaacatg tgagcaaaag gccagcaaaa
      961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1621 tctattcaac aaagccgccg tcccgtcaag tcagcgtaaa tgggtagggg gcttcaaatc
     1681 gtccgctctg ccagtgttac aaccaattaa caaattctga ttagaaaaac tcatcgagca
     1741 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1801 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1861 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1921 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1981 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     2041 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     2101 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     2161 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2221 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2281 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2341 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2401 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     2461 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
     2521 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     2581 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     2641 ggctttcccc cgccgctcta gaactagtgg atccaaataa aacgaaaggc tcagtcgaaa
     2701 gactgggcct ttcgttttat ctgttgtttg tcgcattata cgagacgtcc aggttgggat
     2761 acctgaaaca aaacccatcg tacggccaag gaagtctcca ataactgtga tccaccacaa
     2821 gcgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtcatgc ataatccgca
     2881 cgcatctgga ataaggaagt gccattccgc ctgacct
//
