LOCUS       pSEM254___sgRNA_        2522 bp    DNA        circular     14-AUG-2020
DEFINITION  .
COMMENT     Unpublished reagents. Sonia el Mouridi and Christian
            Froekjaer-Jensen. Synthetic Genome Biology lab. KAUST.
COMMENT     www.wormbuilder.org
COMMENT     MOdular Safe harbor Transgene Insertion (mosTI). 
COMMENT     Split neoR selection
COMMENT     sgRNA for single-copy or targeted extrachromosomal insertion at
            landing site.
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_RNA        190..286
                     /locus_tag="sgRNA scaffold w terminator"
                     /label="sgRNA scaffold w terminator"
                     /ApEinfo_label="sgRNA scaffold w terminator"
                     /ApEinfo_fwdcolor="#9437ff"
                     /ApEinfo_revcolor="#9437ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     48..69
                     /locus_tag="UniF"
                     /label="UniF"
                     /ApEinfo_label="UniF"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1..17
                     /locus_tag="M13F"
                     /label="M13F"
                     /ApEinfo_label="M13F"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(427..443)
                     /locus_tag="M13R"
                     /label="M13R"
                     /ApEinfo_label="M13R"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(371..391)
                     /locus_tag="UniR"
                     /label="UniR"
                     /ApEinfo_label="UniR"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     promoter        70..170
                     /locus_tag="Prpr-1 (short, w 1st G) (Aljohani unpub)"
                     /label="Prpr-1 (short, w 1st G) (Aljohani unpub)"
                     /ApEinfo_label="Prpr-1 (short, w 1st G) (Aljohani unpub)"
                     /ApEinfo_fwdcolor="#72fcd5"
                     /ApEinfo_revcolor="#72fcd5"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1444..2409
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#929292"
                     /ApEinfo_revcolor="#929292"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    615..1203
                     /locus_tag="Ori"
                     /label="Ori"
                     /ApEinfo_label="Ori"
                     /ApEinfo_fwdcolor="#d5d5d5"
                     /ApEinfo_revcolor="#d5d5d5"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    171..189
                     /locus_tag="MosTI protospacer NeoR"
                     /label="MosTI protospacer NeoR"
                     /ApEinfo_label="MosTI protospacer NeoR"
                     /ApEinfo_fwdcolor="#7980ff"
                     /ApEinfo_revcolor="#7980ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
ORIGIN
        1 gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc
       61 gcctgacctc gcctccccgc acactcaaca cagcgagcgg cggaacccgc ggggtgtcgg
      121 ccgctcggac acgctcccgt atataacgac gcggcgcgcg tcaagttgtG CTTCAGACTC
      181 GCAATGTAGG TTTAAGAGCT ATGCTGGAAA CAGCATAGCA AGTTTAAATA AGGCTAGTCC
      241 GTTATCAACT TGAAAAAGTG GCACCGAGTC GGTGCTtttt gttttttgcc cattttttgt
      301 agtttgagac tggaaaaatc gtaaatcaac catagtttta gggcaaaatt ctcgaaattt
      361 ttctaaaaat aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt
      421 gtcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat
      481 tccgcgctat ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat
      541 gtcgctggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1201 caagaagatc ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc
     1261 ccgtcaagtc agcgtaaatg ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga
     1321 gcctgctttt ttgtacaaac ttgttgataa tggcaattca aggatcttca cctagatcct
     1381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1441 cagTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT TTCGTTCATC
     1501 CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA CGGGAGGGCT TACCATCTGG
     1561 CCCCAGTGCT GCAATGATAC CGCGAGAGCC ACGCTCACCG GCTCCAGATT TATCAGCAAT
     1621 AAACCAGCCA GCCGGAAGGG CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT
     1681 CCAGTCTATT AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG
     1741 CAACGTTGTT GCCATTGCTA CAGGCATCGT GGTGTCACGC TCGTCGTTTG GTATGGCTTC
     1801 ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA TCCCCCATGT TGTGCAAAAA
     1861 AGCGGTTAGC TCCTTCGGTC CTCCGATCGT TGTCAGAAGT AAGTTGGCCG CAGTGTTATC
     1921 ACTCATGGTT ATGGCAGCAC TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT
     1981 TTCTGTGACT GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG
     2041 TTGCTCTTGC CCGGCGTCAA TACGGGATAA TACCGCGCCA CATAGCAGAA CTTTAAAAGT
     2101 GCTCATCATT GGAAAACGTT CTTCGGGGCG AAAACTCTCA AGGATCTTAC CGCTGTTGAG
     2161 ATCCAGTTCG ATGTAACCCA CTCGTGCACC CAACTGATCT TCAGCATCTT TTACTTTCAC
     2221 CAGCGTTTCT GGGTGAGCAA AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC
     2281 GACACGGAAA TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA
     2341 GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG
     2401 GGTTCCGCGc acatttcccc gaaaagtgcc agatacctga aacaaaaccc atcgtacggc
     2461 caaggaagtc tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg
     2521 tt     
//
