LOCUS       pKO024.306-107          3081 bp ds-DNA     circular     09-MAY-2025
DEFINITION  .
KEYWORDS    "Construction Method" "Origin" "Parental Vector" "Promoter"
            "Resistance" "Terminator" "accession:addgene_35279_42946" "gRNA"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
FEATURES             Location/Qualifiers
     terminator      90..184
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(215..1009)
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     complement(326..345)
                     /label="Neo-F"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     936..955
                     /label="Neo-R"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     promoter        1271..1300
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        1301..1305
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    1306..1325
                     /label="J306"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_fwdcolor="#800080"
     misc_feature    1326..1337
                     /label="crRNA"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1342..1401
                     /label="tracR"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1375..1399
                     /label="MS2"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1407..1423
                     /label="FWD"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     terminator      1407..1764
                     /label="TrrnB"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      1700..1764
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     misc_feature    1756..1787
                     /label="problematic overhang"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     promoter        1784..1818
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    1819..1838
                     /label="J107"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1839..1880
                     /label="JAD designed chimera RNA 3'UTR"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1885..1921
                     /label="Streptococcus pyogenes tracrRNA terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      1922..2289
                     /label="TrrnB"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1929..1934
                     /label="Bsp120I"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     terminator      2215..2282
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     rep_origin      complement(2475..76)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagAGT ACTgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
      121 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
      181 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
      241 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
      301 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
      361 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
      421 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
      481 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
      541 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
      601 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
      661 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
      721 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
      781 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
      841 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
      901 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
      961 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     1021 ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga
     1081 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     1141 attccgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1201 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggAATT
     1261 CTAAAGATCT TTGACAGCTA GCTCAGTCCT AGGTATAATA CTAGTTTGTG TCCAGAACGC
     1321 TCCGTGTTTT TGTACTCGAA AGAAGCTACA AAGATAAGGC TTCATGCCGA AATCGCGCAC
     1381 ATGAGGATCA CCCATGTGCT TTTTTTGAAG CTTGGGCCCG AACAAAAACT Catctcagaa
     1441 gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc
     1501 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     1561 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     1621 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     1681 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactGG
     1741 GCCTTTCGTT TTATCTGTTG TTTGTCGGTG AACTGGATCT TACTTGACAG CTAGCTCAGT
     1801 CCTAGGTATA ATACTAGTCG GTGTCCTGCG GTTACCAAGT TTTAGAGCTA GAAATAGCAA
     1861 GTTAAAATAA GGCTAGTCCG TTATCAACTT GAAAAAGTGG CACCGAGTCG GTGCTTTTTT
     1921 TGAAGCTTGG GCCCGAACAA AAACTCatct cagaagagga tctgaatagc gccgtcgacc
     1981 atcatcatca tcatcattga gtttaaacgg tctccagctt ggctgttttg gcggatgaga
     2041 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     2101 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     2161 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     2221 atcaaataaa acgaaaggct cagtcgaaag actGGGCCTT TCGTTTTATC TGTTGTTTGT
     2281 CGGTGAACTG GATCCTTACT CGAGTCTAGA CTGCAGgctt cctcgctcac tgactcgctg
     2341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3061 cgcagaaaaa aaggatctca a
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