LOCUS pDGB3_α2-pegRNA-K172R-T4 6958 bp DNA circular SYN 07-NOV-2024 DEFINITION a2_pegRNA-K172R-T4. ACCESSION VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid. REFERENCE 1 (bases 1 to 6958) AUTHORS Johan TITLE Direct Submission JOURNAL Exported Oct 21, 2024 from SnapGene 7.2.1 https://www.snapgene.com FEATURES Location/Qualifiers misc_feature 1..11 /created_by="mamee" /label /note="Geneious type: Golden Braid overhang" promoter 16..334 /modified_by="mamee" /label="StU6 promoter" gRNA 338..357 /modified_by="mamee" /label="20bp Target" Scaffold 358..443 /modified_by="mamee" /label="esgRNA" misc_feature 444..471 /label="Rt template" Change 453..454 /label="S172K" Change 462 /label="Silent SNP" Change 465 /label="Silent SNP" Change 468 /label="Silent SNP" misc_feature 472..482 /label="PBS" Linker 483..490 /modified_by="mamee" /label="Peg-Linker" misc_feature 491..576 /note="TYMV TLS" terminator 577..596 /note="transcription terminator for the S. cerevisiae SUP4 tRNA gene" /label="SUP4 terminator" misc_feature complement(597..607) /created_by="mamee" /label /note="Geneious type: Golden Braid overhang" T-DNA_border 614..638 /note="right border repeat from nopaline C58 T-DNA" /modified_by="mamee" /label="RB" regulatory 1939..2568 /note="Pseudomonas pVS1 plasmid stability protein" /product="plasmid stability protein StaA" /gene="staA" /regulatory_class="other" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pVS1 staA" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; pVS1 staA" /label="pVS1 staA" regulatory 2997..4069 /note="Pseudomonas pVS1 plasmid replication protein" /product="replication protein RepA" /gene="repA" /regulatory_class="other" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pVS1 repA" /Transferred_Similarity="99.91%" /Primary_Match="99.91%; Adjusted-Interval-0:0/0; pVS1 repA" /label="pVS1 repA" rep_origin 4135..4329 /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label="pVS1 oriV" regulatory 4703..4963 /%_Identity=100 /Motif="CGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAG AGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATC AGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAA CGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAG" /annotation_group="pBR322: 7,710 -> 7,970" /modified_by="User" /modified_by="mamee" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pBR322 bom" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; pBR322 bom" /Alternative_Match="99.29%; Adjusted-Interval-0:-30/-150; bom" /label="pBR322 bom" rep_origin complement(4999..5587) /created_by="thjac" /modified_by="mamee" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pColE1 ori" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; pColE1 ori" /Alternative_Match="99.83%; Adjusted-Interval-0:0/0; pUC ori" /label="pColE1 ori" misc_feature complement(5675..6490) /modified_by="mamee" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="KanR" /Transferred_Similarity="98.53%" /Primary_Match="98.53%; Adjusted-Interval-0:0/0; KanR" /label="KanR" /note="Geneious type: Bacterial Selection" T-DNA_border 6923..6947 /note="left border repeat from nopaline C58 T-DNA" /modified_by="mamee" /label="LB" ORIGIN 1 cgtctcagtc aggagtttca cttagttcag ttgcattatg tctttataca cccctaccta 61 gatgattaag ttttacttta gttggtgtga aatggataaa ttctaaaata tgaggtgtgg 121 aatgaaggat tgtcatcaat tagttggccc caaccaagta aaataagaag gccggcccat 181 tacaattaag tcgtcacaca agtgggcttc attgaaacaa gcgcaaaaag gagtccaggc 241 ccgtgttagc gtgaagactc aaccagcgat ttctccctca tcggttgcac agaaaagctg 301 tgtgttgttt atatggcgaa acctaacagt ctgacttgat catggagatg aaatttggtt 361 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg 421 aaaaagtggc accgagtcgg tgcagaccgg accttaacga ccgcgccgca aatttcatct 481 cctctctctc gttagctcgc cagttagcga ggtctgtccc cacacgacag ataatcgggt 541 gcaactcccg cccctcttcc gagggtcatc ggaacctttt tttgtttttt atgtctcgct 601 tgagacgaag ctttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt 661 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg 721 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct 781 atagtgcagt cggcttctga cgttcagtgc agccgtcatc tgaaaacgac atgtcgcaca 841 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt 901 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca 961 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga 1021 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca 1081 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg 1141 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca 1201 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg 1261 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg 1321 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg 1381 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga 1441 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga 1501 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg 1561 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac 1621 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac 1681 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacct 1741 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg 1801 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt 1861 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca 1921 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc 1981 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg 2041 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa 2101 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc 2161 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg 2221 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc 2281 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa 2341 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag 2401 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac 2461 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc 2521 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta 2581 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca 2641 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc 2701 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca 2761 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa 2821 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc 2881 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc 2941 tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga 3001 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga 3061 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg 3121 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc 3181 cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc 3241 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg 3301 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca 3361 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact 3421 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa 3481 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga 3541 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt 3601 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga 3661 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga 3721 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct 3781 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct 3841 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg 3901 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc 3961 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt 4021 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca 4081 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggactct ttcctgtgga 4141 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa 4201 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa 4261 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc 4321 ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg 4381 gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc 4441 aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc 4501 actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg 4561 aaaacctctg acacatgcag ctcccggtga cggtcacagc ttgtctgtaa gcggatgccg 4621 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca 4681 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca 4741 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 4801 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 5161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 5221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 5281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 5341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 5401 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 5461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 5521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5641 acgttaaggg attttggtca tgcattctag gtgattagaa aaactcatcg agcatcaaat 5701 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct 5761 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt 5821 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa 5881 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt 5941 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac 6001 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagtcga aatacgcgat 6061 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca 6121 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt 6181 tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga 6241 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat 6301 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat 6361 acaatcggta gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat 6421 ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt tcccgttgaa 6481 tatggctcat aacagaactt attatttcct tcctcttttc tacagtattt aaagataccc 6541 caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt 6601 aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg 6661 acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca tcgttacaat 6721 caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc 6781 ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga 6841 cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt 6901 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacata acaagctt //