LOCUS       pFH71                   2553 bp    DNA     circular UNA 30-MAY-2019
DEFINITION  Subcloning of sgRNA protospacer/ tRNA-gRNA system with improved
            gRNA backbone/ Position 1.
ACCESSION   urn.local...25-aj37n96
VERSION     urn.local...25-aj37n96
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"
     overhang        73..76
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="CTTG overhang to pICSL90003"
     Start_codon     76
                     /created_by="hahnf"
                     /label="Transcription Start Signal (G)"
     tRNA            77..153
                     /created_by="hahnf"
                     /Color="#ff7400"
                     /Transferred_From="pFH26, pFH27, pFH28"
                     /Transferred_Similarity="100.00%"
                     /label="tRNA Glycine"
     overhang        150..153
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="TGCA"
     misc_feature    complement(156..161)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     CDS             162..739
                     /created_by="koroleva"
                     /Transferred_From="AK001 in pUC57-Kan-mini, AK002 in
                     pUC57-Kan-mini, pAK001 in pUC57-Kan-mini, pAK002 in
                     pUC57-Kan-mini"
                     /Transferred_Similarity="100.00%"
                     /label="LacZ"
     misc_feature    740..745
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     gRNA_Scaffold   748..787
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="improved sgRNA backbone (part)"
     overhang        748..751
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="GTTT"
     overhang        784..787
                     /created_by="hahnf"
                     /label="AAAT"
     misc_feature    complement(789..794)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(850..1464)
                     /vntifkey=21
                     /Transferred_From="pAK-EL-01, pAK-EL-02, pAK-EL-03,
                     pAK-EL-04, pAK-EL-05"
                     /Transferred_Similarity="100.00%"
                     /label="rep(pMB1)"
     CDS             complement(1631..2440)
                     /vntifkey=21
                     /Transferred_From="pAK-EL-01, pAK-EL-02, pAK-EL-03,
                     pAK-EL-04, pAK-EL-05"
                     /Transferred_Similarity="100.00%"
                     /label="KanR"
     primer_bind     2511..2534
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="2492998740745d7af307b-ad34-4a87-87a8-2b
                     dffb2c6aba"
                     /Sequence="GCAATGTAACATCAGAGATTTTGA"
                     /created_by="primer3"
                     /Transferred_From="AK001 in pUC57-Kan-mini"
                     /Transferred_Similarity="100.00%"
                     /label="FH32"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tcgatggtct cacttgaaca aagcaccagt ggtctagtgg tagaatagta ccctgccacg
      121 gtacagaccc gggttcgatt cccggctggt gcattgtctt cgcagctggc acgacaggtt
      181 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      241 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      301 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
      361 ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
      421 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      481 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      541 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      601 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      661 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      721 cttacagaca agctgtgacg aagacaagtt tcagagctat gctggaaaca gcatagcaag
      781 ttgaaattga gaccatcatt gtcacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1561 tttaaattaa aaatgaagtt ttaaatcaag cccaatctga ataatgttac aaccaattaa
     1621 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1681 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1741 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1801 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1861 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     1921 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1981 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     2041 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     2101 aatcaggata ttcttctaat acctggaatg ctgtttttcc ggggatcgca gtggtgagta
     2161 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     2221 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     2281 gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg
     2341 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     2401 ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2461 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2521 atcagagatt ttgagacacg ggccagagct gca
//
