LOCUS       pTBL3428_MTK234_        2118 bp    DNA        circular     09-AUG-2024
DEFINITION  .
ACCESSION   <unknown id>                                                       
VERSION     <unknown id>                                                       
KEYWORDS    "sgRNA Dropout"                                                    
COMMENT     
COMMENT     see https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6765135/
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    986..986
                     /locus_tag="human U6 promoter"
                     /label="human U6 promoter"
                     /ApEinfo_label="human U6 promoter"
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="#00ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    733..736
                     /locus_tag="BsaI"
                     /label="BsaI"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    844..863
                     /locus_tag="lacO1"
                     /label="lacO1"
                     /ApEinfo_label="lacO1"
                     /ApEinfo_fwdcolor="#fb0106"
                     /ApEinfo_revcolor="#fb0106"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1020..1031
                     /locus_tag="modified stem loop"
                     /label="modified stem loop"
                     /ApEinfo_label="modified stem loop"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    938..956
                     /locus_tag="LacO?"
                     /label="LacO?"
                     /ApEinfo_label="LacO?"
                     /ApEinfo_fwdcolor="#fefe0a"
                     /ApEinfo_revcolor="#fefe0a"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(1311..1970)
                     /locus_tag="CamR"
                     /label="CamR"
                     /ApEinfo_label="CamR"
                     /ApEinfo_fwdcolor="#0000ff"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1127..1146
                     /locus_tag="HIS6-insertion"
                     /label="HIS6-insertion"
                     /ApEinfo_label="HIS6-insertion"
                     /ApEinfo_fwdcolor="#72fcd5"
                     /ApEinfo_revcolor="#72fcd5"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(1971..2075)
                     /locus_tag="CamR Promoter"
                     /label="CamR Promoter"
                     /ApEinfo_label="CamR Promoter"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     terminator      complement(1202..1310)
                     /locus_tag="CamR Terminator"
                     /label="CamR Terminator"
                     /ApEinfo_label="CamR Terminator"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     rep_origin      complement(join(2076..2118,1..721))
                     /locus_tag="ColE1"
                     /label="ColE1"
                     /ApEinfo_label="ColE1"
                     /ApEinfo_fwdcolor="#7f7f7f"
                     /ApEinfo_revcolor="#7f7f7f"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     protein_bind    complement(1194..1199)
                     /locus_tag="BsaI(1)"
                     /label="BsaI(1)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     protein_bind    complement(1189..1192)
                     /locus_tag="BsaI(2)"
                     /label="BsaI(2)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     protein_bind    726..731
                     /locus_tag="BsaI(3)"
                     /label="BsaI(3)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1008..1018
                     /locus_tag="modified stem loop(1)"
                     /label="modified stem loop(1)"
                     /ApEinfo_label="modified stem loop"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1127..1146
                     /locus_tag="longpeg"
                     /label="longpeg"
                     /ApEinfo_label="longpeg"
                     /ApEinfo_fwdcolor="#ff84ff"
                     /ApEinfo_revcolor="#ff84ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1093..1126
                     /locus_tag="longpeg(1)"
                     /label="longpeg(1)"
                     /ApEinfo_label="longpeg"
                     /ApEinfo_fwdcolor="#ff84ff"
                     /ApEinfo_revcolor="#ff84ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1033..1092
                     /locus_tag="modified stem loop(2)"
                     /label="modified stem loop(2)"
                     /ApEinfo_label="modified stem loop"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1161..1165
                     /locus_tag="modified stem loop(3)"
                     /label="modified stem loop(3)"
                     /ApEinfo_label="modified stem loop"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1004..1006
                     /locus_tag="site3sgRNA"
                     /label="site3sgRNA"
                     /ApEinfo_label="site3sgRNA"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="cyan"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1147..1160
                     /locus_tag="PBS"
                     /label="PBS"
                     /ApEinfo_label="PBS"
                     /ApEinfo_fwdcolor="#9c5aff"
                     /ApEinfo_revcolor="#9c5aff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
ORIGIN
        1 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
       61 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      721 tagtcggtct cAaacgGAGG GCCTATTTCC CATGATTCCT TCATATTTGC ATATACGATA
      781 CAAGGCTGTT AGAGAGATAA TTAGAATTAA TTTGACTGTA AACACAAAGA TATTAGTACA
      841 AAAAATTGTG AGCGcTcACA ATTATTTCTT GGGTAGTTTG CAGTTTTAAA ATTATGTTTT
      901 AAAATGGACT ATCATATGCT TACCGTAACT TGAAAGTAAT TGTGAGCGCT CACAATTATA
      961 TATCTTGTGG AAAGGACGAA ACACCgACTG GGCCATCTAT CATTGAGTTT AAGAGCTAAG
     1021 CTGGAAACAG CATAGCAAGT TTAAATAAGG CTAGTCCGTT ATCAACTTGA AAAAGTGGCA
     1081 CCGAGTCGGT GCTGGAGGAA GCAGGGCTTC CTTTCCTCTG CCATCAATGA TGGTGATGAT
     1141 GGCctaATGA TAGATGGCCC TTTTTccatc ttctagaaag actggagtgc tgtgagacca
     1201 gaccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     1261 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
     1321 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     1381 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     1441 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     1501 tcaaaactgg tgaaactcac ccagggattg gctgaaacga aaaacatatt ctcaataaac
     1561 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     1621 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     1681 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     1741 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     1801 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     1861 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     1921 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     1981 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     2041 tggtgaaagt tggaacctct tacgtgcccg atcaatcatg accaaaatcc cttaacgtga
     2101 gttttcgttc cactgagc    
//
