LOCUS pCDB-Cas9 8957 bp DNA circular UNA 23-FEB-2023 DEFINITION . ACCESSION urn.local...b-cixsr3m VERSION urn.local...b-cixsr3m KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers attL1 1..100 /Color="#00cccc" /created_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attL1" /Transferred_Similarity="99.00%" /Primary_Match="99.00%; Adjusted-Interval-0:0/0; attL1" /label="attL1" promoter 107..970 /regulatory_class="promoter" /gene="CaMV 35S" /note="internal BsaI recognition site removed" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCaMV 35S - Golden Gate" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; p35S (pCaMV 35S - Golden Gate)" /label="p35S" Linker 975..998 /gene="B-dummy" /note="default module for GreenGate" /modified_by="mamee" /label="B-dummy 5'UTR" CDS 1005..5108 /modified_by="mamee" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="AtCas9" /Transferred_Similarity="100.00%" /Adjusted_Transferred_Similarity="99.13%" /Primary_Match="100.00%; Adjusted-Interval-0:0/-36 (adjusted to 99.13%; Adjusted-Interval-0:0/0); AtCas9" /label="AtCAS9" NLS 5121..5141 /created_by="warddecaestecker" /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.2 '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 (NLS SV40)" /label="SV40" Linker 5149..5194 /gene="D-dummy" /note="default module for GreenGate" /modified_by="mamee" /label="D-dummy 3'UTR" terminator 5199..5845 /regulatory_class="terminator" /note="RBCS terminator (from Pisum sativum)" /modified_by="mamee" /label="tRbcS" promoter 5852..6239 /vntifkey=29 /modified_by="thjac" /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="AtU6-26" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; AtU6-26" /label="AtU6-26" Scaffold 6258..6340 /%_Identity=100 /Motif="GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC TTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTT" /annotation_group="Scaffold: 451 -> 533" /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="SpCas9 Scaffold" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; Scaffold (SpCas9 Scaffold)" /label="Scaffold" attL2 complement(6346..6445) /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="attL2" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL2" /label="attL2" primer complement(6483..6503) /label="GWM13R" promoter 6564..6614 /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 (pCG034_pKan_Kan-CBE)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG040_ABE-Nuc-NLS_Kan -reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG043_Kan-ABE-Dest_co ntrol_ccdB)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG046_ABE-LbCas12a-NL S_Kan_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG005_pGGD-pKan-KanR- E)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG006_pGGA-Kan_promot er-B)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG035_pKan_Kan)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG039_ABE-N7-NLS_Kan- reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG041_ABE-SV40-NLS_Ka n-reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG054_Kan_CBE_control )" /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 (KanR)" /label="kan" misc_feature 6615..7424 /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; KanR" /label="KanR" /note="Geneious type: Bacterial Selection" rep_origin 7571..8159 /created_by="thjac" /modified_by="mamee" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ColE1/pMB1/pBR322/pUC Ori (high-copy-number ColE1/pMB1/pBR322/pUC origin of replication)" /label="ColE1/pMB1/pBR322/pUC Ori" misc_feature 8708..8875 /Original_Bases /label /note="Geneious type: Editing History Insertion" primer 8926..8946 /label="GWM13F" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa 61 tgctttttta taatgccaac tttgtacaaa aaagcaggct taacctccga tccattcggt 121 ccccagatta gccttttcaa tttcagaaag aatgctaacc cacagatggt tagagaggct 181 tacgcagcac gtctcatcaa gacgatctac ccgagcaata atctccagga aatcaaatac 241 cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaactgcat caagaacaca 301 gagaaagata tatttctcaa gatcagaagt actattccag tatggacgat tcaaggcttg 361 cttcacaaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt tcccactgaa 421 tcaaaggcca tggagtcaaa gattcaaata gaggacctaa cagaactcgc cgtaaagact 481 ggcgaacagt tcatacagag tctcttacga ctcaatgaca agaagaaaat cttcgtcaac 541 atggtggagc acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac 601 caaagggcaa ttgagacttt tcaacaaagg gtaatatccg gaaacctcct cggattccat 661 tgcccagcta tctgtcactt tattgtgaag atagtggaaa aggaaggtgg ctcctacaaa 721 tgccatcatt gcgataaagg aaaggccatc gttgaagatg cctctgccga cagtggtccc 781 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct 841 tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac 901 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gagaacacgg 961 gggacgagct aacagtattc agtcgactgg taccaacagg ctccatggat aagaagtact 1021 ctatcggact cgatatcgga actaactctg tgggatgggc tgtgatcacc gatgagtaca 1081 aggtgccatc taagaagttc aaggttctcg gaaacaccga taggcactct atcaagaaaa 1141 accttatcgg tgctctcctc ttcgattctg gtgaaactgc tgaggctacc agactcaaga 1201 gaaccgctag aagaaggtac accagaagaa agaacaggat ctgctacctc caagagatct 1261 tctctaacga gatggctaaa gtggatgatt cattcttcca caggctcgaa gagtcattcc 1321 tcgtggaaga agataagaag cacgagaggc accctatctt cggaaacatc gttgatgagg 1381 tggcatacca cgagaagtac cctactatct accacctcag aaagaagctc gttgattcta 1441 ctgataaggc tgatctcagg ctcatctacc tcgctctcgc tcacatgatc aagttcagag 1501 gacacttcct catcgagggt gatctcaacc ctgataactc tgatgtggat aagttgttca 1561 tccagctcgt gcagacctac aaccagcttt tcgaagagaa ccctatcaac gcttcaggtg 1621 tggatgctaa ggctatcctc tctgctaggc tctctaagtc aagaaggctt gagaacctca 1681 ttgctcagct ccctggtgag aagaagaacg gacttttcgg aaacttgatc gctctctctc 1741 tcggactcac ccctaacttc aagtctaact tcgatctcgc tgaggatgca aagctccagc 1801 tctcaaagga tacctacgat gatgatctcg ataacctcct cgctcagatc ggagatcagt 1861 acgctgattt gttcctcgct gctaagaacc tctctgatgc tatcctcctc agtgatatcc 1921 tcagagtgaa caccgagatc accaaggctc cactctcagc ttctatgatc aagagatacg 1981 atgagcacca ccaggatctc acacttctca aggctcttgt tagacagcag ctcccagaga 2041 agtacaaaga gattttcttc gatcagtcta agaacggata cgctggttac atcgatggtg 2101 gtgcatctca agaagagttc tacaagttca tcaagcctat cctcgagaag atggatggaa 2161 ccgaggaact cctcgtgaag ctcaatagag aggatcttct cagaaagcag aggaccttcg 2221 ataacggatc tatccctcat cagatccacc tcggagagtt gcacgctatc cttagaaggc 2281 aagaggattt ctacccattc ctcaaggata acagggaaaa gattgagaag attctcacct 2341 tcagaatccc ttactacgtg ggacctctcg ctagaggaaa ctcaagattc gcttggatga 2401 ccagaaagtc tgaggaaacc atcacccctt ggaacttcga agaggtggtg gataagggtg 2461 ctagtgctca gtctttcatc gagaggatga ccaacttcga taagaacctt ccaaacgaga 2521 aggtgctccc taagcactct ttgctctacg agtacttcac cgtgtacaac gagttgacca 2581 aggttaagta cgtgaccgag ggaatgagga agcctgcttt tttgtcaggt gagcaaaaga 2641 aggctatcgt tgatctcttg ttcaagacca acagaaaggt gaccgtgaag cagctcaaag 2701 aggattactt caagaaaatc gagtgcttcg attcagttga gatttctggt gttgaggata 2761 ggttcaacgc atctctcgga acctaccacg atctcctcaa gatcattaag gataaggatt 2821 tcttggataa cgaggaaaac gaggatatct tggaggatat cgttcttacc ctcaccctct 2881 ttgaagatag agagatgatt gaagaaaggc tcaagaccta cgctcatctc ttcgatgata 2941 aggtgatgaa gcagttgaag agaagaagat acactggttg gggaaggctc tcaagaaagc 3001 tcattaacgg aatcagggat aagcagtctg gaaagacaat ccttgatttc ctcaagtctg 3061 atggattcgc taacagaaac ttcatgcagc tcatccacga tgattctctc acctttaaag 3121 aggatatcca gaaggctcag gtttcaggac agggtgatag tctccatgag catatcgcta 3181 acctcgctgg atctcctgca atcaagaagg gaatcctcca gactgtgaag gttgtggatg 3241 agttggtgaa ggtgatggga aggcataagc ctgagaacat cgtgatcgaa atggctagag 3301 agaaccagac cactcagaag ggacagaaga actctaggga aaggatgaag aggatcgagg 3361 aaggtatcaa agagcttgga tctcagatcc tcaaagagca ccctgttgag aacactcagc 3421 tccagaatga gaagctctac ctctactacc tccagaacgg aagggatatg tatgtggatc 3481 aagagttgga tatcaacagg ctctctgatt acgatgttga tcatatcgtg ccacagtcat 3541 tcttgaagga tgattctatc gataacaagg tgctcaccag gtctgataag aacaggggta 3601 agagtgataa cgtgccaagt gaagaggttg tgaagaaaat gaagaactat tggaggcagc 3661 tcctcaacgc taagctcatc actcagagaa agttcgataa cttgactaag gctgagaggg 3721 gaggactctc tgaattggat aaggcaggat tcatcaagag gcagcttgtg gaaaccaggc 3781 agatcactaa gcacgttgca cagatcctcg attctaggat gaacaccaag tacgatgaga 3841 acgataagtt gatcagggaa gtgaaggtta tcaccctcaa gtcaaagctc gtgtctgatt 3901 tcagaaagga tttccaattc tacaaggtga gggaaatcaa caactaccac cacgctcacg 3961 atgcttacct taacgctgtt gttggaaccg ctctcatcaa gaagtatcct aagctcgagt 4021 cagagttcgt gtacggtgat tacaaggtgt acgatgtgag gaagatgatc gctaagtctg 4081 agcaagagat cggaaaggct accgctaagt atttcttcta ctctaacatc atgaatttct 4141 tcaagaccga gattaccctc gctaacggtg agatcagaaa gaggccactc atcgagacaa 4201 acggtgaaac aggtgagatc gtgtgggata agggaaggga tttcgctacc gttagaaagg 4261 tgctctctat gccacaggtg aacatcgtta agaaaaccga ggtgcagacc ggtggattct 4321 ctaaagagtc tatcctccct aagaggaact ctgataagct cattgctagg aagaaggatt 4381 gggaccctaa gaaatacggt ggtttcgatt ctcctaccgt ggcttactct gttctcgttg 4441 tggctaaggt tgagaaggga aagagtaaga agctcaagtc tgttaaggaa cttctcggaa 4501 tcactatcat ggaaaggtca tctttcgaga agaacccaat cgatttcctc gaggctaagg 4561 gatacaaaga ggttaagaag gatctcatca tcaagctccc aaagtactca ctcttcgaac 4621 tcgagaacgg tagaaagagg atgctcgctt ctgctggtga gcttcaaaag ggaaacgagc 4681 ttgctctccc atctaagtac gttaactttc tttacctcgc ttctcactac gagaagttga 4741 agggatctcc agaagataac gagcagaagc aacttttcgt tgagcagcac aagcactact 4801 tggatgagat catcgagcag atctctgagt tctctaaaag ggtgatcctc gctgatgcaa 4861 acctcgataa ggtgttgtct gcttacaaca agcacagaga taagcctatc agggaacagg 4921 cagagaacat catccatctc ttcaccctta ccaacctcgg tgctcctgct gctttcaagt 4981 acttcgatac aaccatcgat aggaagagat acacctctac caaagaagtg ctcgatgcta 5041 ccctcatcca tcagtctatc actggactct acgagactag gatcgatctc tcacagctcg 5101 gtggtgattc aagggctgat cctaagaaga agaggaaggt ttgatcaggt ggatcctaga 5161 taacctttac cttcattccc tgtaaattgg taccctgcaa agctttcgtt cgtatcatcg 5221 gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca ccagaatcct 5281 actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt tgttgtgctt 5341 gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg 5401 agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat tatttgtttt 5461 ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac attttgtttt 5521 gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag gagtaaaaca 5581 cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca gacctagaaa 5641 agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt tatgaacttt 5701 cctttatgta attttccaga atccttgtca gattctaatc attgctttat aattatagtt 5761 atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta tgacttgcca 5821 attgattgac aacatgcatc aatcgactac tctttttttc ttcttcttcg ttcatacagt 5881 ttttttttgt ttatcagctt acattttctt gaaccgtagc tttcgttttc ttctttttaa 5941 ctttccattc ggagtttttg tatcttgttt catagtttgt cccaggatta gaatgattag 6001 gcatcgaacc ttcaagaatt tgattgaata aaacatcttc attcttaaga tatgaagata 6061 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag 6121 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca 6181 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattgg 6241 gagacccgag ggtctctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt 6301 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gtattaccca gctttcttgt 6361 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta 6421 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt 6481 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca 6541 ttgcacaaga taaaataata tcatcatgaa caataaaact gtctgcttac ataaacagta 6601 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca 6661 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg 6721 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca 6781 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat 6841 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca 6901 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg 6961 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta 7021 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata 7081 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag 7141 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg 7201 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg 7261 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg 7321 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata 7381 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt 7441 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa 7501 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca 7561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 7621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 7681 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 7741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 7801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 7861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 7921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 7981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 8041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 8101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 8161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 8221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 8281 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 8341 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 8401 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg 8461 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt 8521 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt 8581 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa 8641 acaacagata aaacgaaagg cccagacttc cgactgagcc tttcgtttta tttgatgcct 8701 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa 8761 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa 8821 acgaaaggcc cagacttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta 8881 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca 8941 gtcttaagct cgggccc //