LOCUS       pEN-L4-pZmUBI-Cas9PTA-G7T-R1        9165 bp    DNA     circular     23-FEB-2023
DEFINITION  .
ACCESSION   p00112
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     attL4           1..97
                     /created_by="mamee"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attL4"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:lr-eztz0t4"">attL4</a>"
     promoter        104..2090
                     /modified_by="thjac"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pZmUBIL"
                     /Transferred_Similarity="99.95%"
                     /Primary_Match="99.95%; Adjusted-Interval-0:0/0; ZmUBIL
                     (<a href=""urn:local:.:6u-dgycryx"">pZmUBIL</a>)"
                     /Alternative_Match="99.90%; Adjusted-Interval-0:0/1;
                     ZmUBIL.2 (<a
                     href=""urn:local:.:6s-dgycryx"">pZmUBIL.2</a>)"
                     /label="ZmUBIL"
     misc_feature    910
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     intron          1082..2091
                     /created_by="lapau"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pZmUBIL.2"
                     /Transferred_Similarity="99.90%"
                     /Primary_Match="99.90%; Adjusted-Interval-0:0/0; ZmUBI
                     intron (<a href=""urn:local:.:6s-dgycryx"">pZmUBIL.2</a>)"
                     /label="ZmUBI intron"
     CDS             2125..6240
                     /label="CAS9"
     NLS             6241..6261
                     /created_by="warddecaestecker"
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on protein similarity"
                     /Transferred_From="NLS SV40"
                     /Transferred_Translation_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SV40 (<a
                     href=""urn:local:.:9x-dgycryz"">NLS SV40</a>)"
                     /label="SV40"
     Linker          6269..6314
                     /created_by="wacae"
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Linker"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6m-dgycryx"">Linker</a>"
     terminator      6319..6590
                     /label="tG7"
     attR1           6620..6776
                     /created_by="mamee"
                     /Color="#00cccc"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attR1"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:y-f0o11e8"">attR1</a>"
     primer          complement(6836..6856)
                     /label="GWM13R"
     promoter        6917..6967
                     /created_by="chgai"
                     /X-Transferred_From="pCG034_pKan_Kan-CBE"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ll-ch8iv1b"">pCG034_pKan_Kan-CBE</a
                     >)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lo-ch8iv1b"">pCG040_ABE-Nuc-NLS_Kan
                     -reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lr-ch8iv1b"">pCG043_Kan-ABE-Dest_co
                     ntrol_ccdB</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lu-ch8iv1c"">pCG046_ABE-LbCas12a-NL
                     S_Kan_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:kx-ch8iv19"">pCG005_pGGD-pKan-KanR-
                     E</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ky-ch8iv19"">pCG006_pGGA-Kan_promot
                     er-B</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lm-ch8iv1b"">pCG035_pKan_Kan</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ln-ch8iv1b"">pCG039_ABE-N7-NLS_Kan-
                     reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lp-ch8iv1b"">pCG041_ABE-SV40-NLS_Ka
                     n-reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lw-ch8iv1c"">pCG054_Kan_CBE_control
                     </a>)"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KanR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (<a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>)"
                     /label="kan"
     misc_feature    6968..7777
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KanR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>"
                     /label="KanR"
                     /note="Geneious type: Bacterial Selection"
     rep_origin      7924..8512
                     /created_by="thjac"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of  replication"
                     /Transferred_Similarity="99.49%"
                     /Primary_Match="99.49%; Adjusted-Interval-0:0/0;
                     ColE1/pMB1/pBR322/pUC Ori (<a
                     href=""urn:local:.:8d-dk4mzop"">high-copy-number
                     ColE1/pMB1/pBR322/pUC origin of  replication</a>)"
                     /label="ColE1/pMB1/pBR322/pUC Ori"
     primer          9111..9131
                     /label="GWM13F"
ORIGIN      
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa
       61 tgctttttta taatgccaac tttgtataga aaagttgcta cctgtgcagc gtgacccggt
      121 cgtgcccctc tctagagata atgagcattg catgtctaag ttataaaaaa ttaccacata
      181 ttttttttgt cacacttgtt tgaagtgcag tttatctatc tttatacata tatttaaact
      241 ttactctacg aataatataa tctatagtac tacaataata tcagtgtttt agagaatcat
      301 ataaatgaac agttagacat ggtctaaagg acaattgagt attttgacaa caggactcta
      361 cagttttatc tttttagtgt gcatgtgttc tccttttttt ttgcaaatag cttcacctat
      421 ataatacttc atccatttta ttagtacatc catttagggt ttagggttaa tggtttttat
      481 agactaattt ttttagtaca tctattttat tctattttag cctctaaatt aagaaaacta
      541 aaactctatt ttagtttttt tatttaataa tttagatata aaatagaata aaataaagtg
      601 actaaaaatt aaacaaatac cctttaagaa attaaaaaaa ctaaggaaac atttttcttg
      661 tttcgagtag ataatgccag cctgttaaac gccgtcgacg agtctaacgg acaccaacca
      721 gcgaaccagc agcgtcgcgt cgggccaagc gaagcagacg gcacggcatc tctgtcgctg
      781 cctctggacc cctctcgaga gttccgctcc accgttggac ttgctccgct gtcggcatcc
      841 agaaattgcg tggcggagcg gcagacgtga gccggcacgg caggcggcct cctcctcctc
      901 tcacggcacc ggcagctacg ggggattcct ttcccaccgc tccttcgctt tcccttcctc
      961 gcccgccgta ataaatagac accccctcca caccctcttt ccccaacctc gtgttgttcg
     1021 gagcgcacac acacacaacc agatctcccc caaatccacc cgtcggcacc tccgcttcaa
     1081 ggtacgccgc tcgtcctccc cccccccccc tctctacctt ctctagatcg gcgttccggt
     1141 ccatggttag ggcccggtag ttctacttct gttcatgttt gtgttagatc cgtgtttgtg
     1201 ttagatccgt gctgctagcg ttcgtacacg gatgcgacct gtacgtcaga cacgttctga
     1261 ttgctaactt gccagtgttt ctctttgggg aatcctggga tggctctagc cgttccgcag
     1321 acgggatcga tttcatgatt ttttttgttt cgttgcatag ggtttggttt gcccttttcc
     1381 tttatttcaa tatatgccgt gcacttgttt gtcgggtcat cttttcatgc ttttttttgt
     1441 cttggttgtg atgatgtggt ctggttgggc ggtcgttcta gatcggagta gaattctgtt
     1501 tcaaactacc tggtggattt attaattttg gatctgtatg tgtgtgccat acatattcat
     1561 agttacgaat tgaagatgat ggatggaaat atcgatctag gataggtata catgttgatg
     1621 cgggttttac tgatgcatat acagagatgc tttttgttcg cttggttgtg atgatgtggt
     1681 gtggttgggc ggtcgttcat tcgttctaga tcggagtaga atactgtttc aaactacctg
     1741 gtgtatttat taattttgga actgtatgtg tgtgtcatac atcttcatag ttacgagttt
     1801 aagatggatg gaaatatcga tctaggatag gtatacatgt tgatgtgggt tttactgatg
     1861 catatacatg atggcatatg cagcatctat tcatatgctc taaccttgag tacctatcta
     1921 ttataataaa caagtatgtt ttataattat tttgatcttg atatacttgg atgatggcat
     1981 atgcagcagc tatatgtgga tttttttagc cctgccttca tacgctattt atttgcttgg
     2041 tactgtttct tttgtcgatg ctcaccctgt tgtttggtgt tacttctgca aacagtattc
     2101 agtcgactgg taccaacagg ctccatggat aagaagtact ctatcggact cgatatcgga
     2161 actaactctg tgggatgggc tgtgatcacc gatgagtaca aggtgccatc taagaagttc
     2221 aaggttctcg gaaacaccga taggcactct atcaagaaaa accttatcgg tgctctcctc
     2281 ttcgattctg gtgaaactgc tgaggctacc agactcaaga gaaccgctag aagaaggtac
     2341 accagaagaa agaacaggat ctgctacctc caagagatct tctctaacga gatggctaaa
     2401 gtggatgatt cattcttcca caggctcgaa gagtcattcc tcgtggaaga agataagaag
     2461 cacgagaggc accctatctt cggaaacatc gttgatgagg tggcatacca cgagaagtac
     2521 cctactatct accacctcag aaagaagctc gttgattcta ctgataaggc tgatctcagg
     2581 ctcatctacc tcgctctcgc tcacatgatc aagttcagag gacacttcct catcgagggt
     2641 gatctcaacc ctgataactc tgatgtggat aagttgttca tccagctcgt gcagacctac
     2701 aaccagcttt tcgaagagaa ccctatcaac gcttcaggtg tggatgctaa ggctatcctc
     2761 tctgctaggc tctctaagtc aagaaggctt gagaacctca ttgctcagct ccctggtgag
     2821 aagaagaacg gacttttcgg aaacttgatc gctctctctc tcggactcac ccctaacttc
     2881 aagtctaact tcgatctcgc tgaggatgca aagctccagc tctcaaagga tacctacgat
     2941 gatgatctcg ataacctcct cgctcagatc ggagatcagt acgctgattt gttcctcgct
     3001 gctaagaacc tctctgatgc tatcctcctc agtgatatcc tcagagtgaa caccgagatc
     3061 accaaggctc cactctcagc ttctatgatc aagagatacg atgagcacca ccaggatctc
     3121 acacttctca aggctcttgt tagacagcag ctcccagaga agtacaaaga gattttcttc
     3181 gatcagtcta agaacggata cgctggttac atcgatggtg gtgcatctca agaagagttc
     3241 tacaagttca tcaagcctat cctcgagaag atggatggaa ccgaggaact cctcgtgaag
     3301 ctcaatagag aggatcttct cagaaagcag aggaccttcg ataacggatc tatccctcat
     3361 cagatccacc tcggagagtt gcacgctatc cttagaaggc aagaggattt ctacccattc
     3421 ctcaaggata acagggaaaa gattgagaag attctcacct tcagaatccc ttactacgtg
     3481 ggacctctcg ctagaggaaa ctcaagattc gcttggatga ccagaaagtc tgaggaaacc
     3541 atcacccctt ggaacttcga agaggtggtg gataagggtg ctagtgctca gtctttcatc
     3601 gagaggatga ccaacttcga taagaacctt ccaaacgaga aggtgctccc taagcactct
     3661 ttgctctacg agtacttcac cgtgtacaac gagttgacca aggttaagta cgtgaccgag
     3721 ggaatgagga agcctgcttt tttgtcaggt gagcaaaaga aggctatcgt tgatctcttg
     3781 ttcaagacca acagaaaggt gaccgtgaag cagctcaaag aggattactt caagaaaatc
     3841 gagtgcttcg attcagttga gatttctggt gttgaggata ggttcaacgc atctctcgga
     3901 acctaccacg atctcctcaa gatcattaag gataaggatt tcttggataa cgaggaaaac
     3961 gaggatatct tggaggatat cgttcttacc ctcaccctct ttgaagatag agagatgatt
     4021 gaagaaaggc tcaagaccta cgctcatctc ttcgatgata aggtgatgaa gcagttgaag
     4081 agaagaagat acactggttg gggaaggctc tcaagaaagc tcattaacgg aatcagggat
     4141 aagcagtctg gaaagacaat ccttgatttc ctcaagtctg atggattcgc taacagaaac
     4201 ttcatgcagc tcatccacga tgattctctc acctttaaag aggatatcca gaaggctcag
     4261 gtttcaggac agggtgatag tctccatgag catatcgcta acctcgctgg atctcctgca
     4321 atcaagaagg gaatcctcca gactgtgaag gttgtggatg agttggtgaa ggtgatggga
     4381 aggcataagc ctgagaacat cgtgatcgaa atggctagag agaaccagac cactcagaag
     4441 ggacagaaga actctaggga aaggatgaag aggatcgagg aaggtatcaa agagcttgga
     4501 tctcagatcc tcaaagagca ccctgttgag aacactcagc tccagaatga gaagctctac
     4561 ctctactacc tccagaacgg aagggatatg tatgtggatc aagagttgga tatcaacagg
     4621 ctctctgatt acgatgttga tcatatcgtg ccacagtcat tcttgaagga tgattctatc
     4681 gataacaagg tgctcaccag gtctgataag aacaggggta agagtgataa cgtgccaagt
     4741 gaagaggttg tgaagaaaat gaagaactat tggaggcagc tcctcaacgc taagctcatc
     4801 actcagagaa agttcgataa cttgactaag gctgagaggg gaggactctc tgaattggat
     4861 aaggcaggat tcatcaagag gcagcttgtg gaaaccaggc agatcactaa gcacgttgca
     4921 cagatcctcg attctaggat gaacaccaag tacgatgaga acgataagtt gatcagggaa
     4981 gtgaaggtta tcaccctcaa gtcaaagctc gtgtctgatt tcagaaagga tttccaattc
     5041 tacaaggtga gggaaatcaa caactaccac cacgctcacg atgcttacct taacgctgtt
     5101 gttggaaccg ctctcatcaa gaagtatcct aagctcgagt cagagttcgt gtacggtgat
     5161 tacaaggtgt acgatgtgag gaagatgatc gctaagtctg agcaagagat cggaaaggct
     5221 accgctaagt atttcttcta ctctaacatc atgaatttct tcaagaccga gattaccctc
     5281 gctaacggtg agatcagaaa gaggccactc atcgagacaa acggtgaaac aggtgagatc
     5341 gtgtgggata agggaaggga tttcgctacc gttagaaagg tgctctctat gccacaggtg
     5401 aacatcgtta agaaaaccga ggtgcagacc ggtggattct ctaaagagtc tatcctccct
     5461 aagaggaact ctgataagct cattgctagg aagaaggatt gggaccctaa gaaatacggt
     5521 ggtttcgatt ctcctaccgt ggcttactct gttctcgttg tggctaaggt tgagaaggga
     5581 aagagtaaga agctcaagtc tgttaaggaa cttctcggaa tcactatcat ggaaaggtca
     5641 tctttcgaga agaacccaat cgatttcctc gaggctaagg gatacaaaga ggttaagaag
     5701 gatctcatca tcaagctccc aaagtactca ctcttcgaac tcgagaacgg tagaaagagg
     5761 atgctcgctt ctgctggtga gcttcaaaag ggaaacgagc ttgctctccc atctaagtac
     5821 gttaactttc tttacctcgc ttctcactac gagaagttga agggatctcc agaagataac
     5881 gagcagaagc aacttttcgt tgagcagcac aagcactact tggatgagat catcgagcag
     5941 atctctgagt tctctaaaag ggtgatcctc gctgatgcaa acctcgataa ggtgttgtct
     6001 gcttacaaca agcacagaga taagcctatc agggaacagg cagagaacat catccatctc
     6061 ttcaccctta ccaacctcgg tgctcctgct gctttcaagt acttcgatac aaccatcgat
     6121 aggaagagat acacctctac caaagaagtg ctcgatgcta ccctcatcca tcagtctatc
     6181 actggactct acgagactag gatcgatctc tcacagctcg gtggtgattc aagggctgat
     6241 cctaagaaga agaggaaggt ttgatcaggt ggatcctaga taacctttac cttcattccc
     6301 tgtaaattgg taccctgccc ccgatgagct aagctagcta tatcatcaat ttatgtatta
     6361 cacataatat cgcactcagt ctttcatcta cggcaatgta ccagctgata taatcagtta
     6421 ttgaaatatt tctgaattta aacttgcatc aataaattta tgtttttgct tggactataa
     6481 tacctgactt gttattttat caataaatat ttaaactata tttctttcaa gatgggaatt
     6541 aacatctaca aattgccttt tcttatcgac catgtactcg acggccgagt actaatagaa
     6601 ttcgatatta tttagtatgc aagtttgtac aaaaaagttg aacgagaaac gtaaaatgat
     6661 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     6721 acacaacata tgcagtcact atgaatcaac tacttagatg gtattagtga cctgtagaat
     6781 tcgagctcta gagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt
     6841 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     6901 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     6961 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     7021 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     7081 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     7141 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     7201 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     7261 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     7321 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     7381 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     7441 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     7501 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     7561 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     7621 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     7681 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     7741 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     7801 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     7861 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7921 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7981 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8041 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     8101 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8161 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8221 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8281 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     8341 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8401 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8461 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8521 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8581 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     8641 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     8701 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     8761 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     8821 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
     8881 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     8941 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     9001 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
     9061 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
     9121 ggccagtctt aagctcgggc ccgcgttaac gctaccatgg agctc
//
