LOCUS pGG-UmpRbcS-BleR-E2A-B-ccdB-CmR-G-UmtRbcS 5733 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.p-cn2ko84 VERSION urn.local.jonasblomme.p-cn2ko84 KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/. FEATURES Location/Qualifiers promoter 1..546 /regulatory_class="promoter" /note="Ulva RbcS promoter" /label="UmpRbcS" CDS join(551..558,1056..1419) /gene="ble" /note="derived from Streptoalloteichus hindustanus, adapted to Chlamydomonas reinhardtii and Ulva mutabilis codon usage" /codon_start=1 /transl_table=11 /product="bleomycin resistance protein" /protein_id="ABY50194.1" /db_xref="GI:163962062" /translation="MAKITSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW GREFALRDPAGNCVHFVAEEQD" intron 559..1055 /label="RbcsI" CDS 1420..1488 /label="E2A" GG complement(1494..1504) /created_by="thjac" /modified_by="thjac" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="B" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; B" /label="B" CDS complement(1511..1816) /vntifkey=4 /label="ccdB" UTR complement(1817..2135) /Mismatches=1 /%_Identity="99.68652037617555" /Motif="AAGCCAGATAACAGTATGCGTATTTGCGCGCTGATTTTTGCGGTATAAGA ATATATACTGATATGTATACCCGAAGTATGTCAAAAGGAGGTATGCTATGAAGCAGCG TATTACAGTGACAGTTGACAGCGACAGCTATCAGTTGCTCAAGGCATATATGATGTCA ATATCTCCGGTCTGGTAAGCACAACCATGCAGAATGAAGCCCGTCGTCTGCGTGCCGA ACGCTGGAAAGCGGAAAATCAGGAAGGGATGGCTGAGGTCGCCCGGTTTATTGAAATG AACGGCTCTTTTGCTGACGAGAACAGGGGCTGGTGAA" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="ccdB" /Transferred_Similarity="99.37%" /Primary_Match="99.37%; Adjusted-Interval-0:0/0; ccdB 5'UTR (ccdB)" /label="ccdB 5'UTR" misc_feature complement(2158..2817) /Transferred_From="cmR" /Transferred_Similarity="99.85%" /modified_by="mamee" /note="Derived using Geneious Prime 2023.0.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="14 CmR" /Transferred_Similarity="99.85%" /Primary_Match="99.85%; Adjusted-Interval-0:0/0; CmR (14 CmR)" /Alternative_Match="99.85%; Adjusted-Interval-0:0/0; CmR" /label="CmR" /note="Geneious type: Bacterial Selection" promoter complement(2871..2901) /note="Lac promoter of Escherichia coli containing 2 base pair mutations" /regulatory_class="promoter" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /X-Transferred_From="lacUV5" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacUV5" /note="Derived using Geneious Prime 2023.0.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="14 CmR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacUV5 (14 CmR)" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; lacUV5 (CmR)" /label="lacUV5" GG 2926..2936 /%_Identity=100 /Motif="GGTCTCAGTAT" /modified_by="thjac" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="G" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; G overlap (G)" /modified_by="mamee" /label="G" terminator 2945..3333 /regulatory_class="terminator" /note="Ulva RbcS terminator" /modified_by="jonasblomme" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="UmtRbcS" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; UmtRbcS" /label="UmtRbcS" Linker 3338..3386 /created_by="wacae" /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="Linker II" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; Linker II" /label="Linker II" primer complement(3423..3443) /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCDB-Cas9 - GWM13R" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13R (pCDB-Cas9 - GWM13R)" /label="GWM13R" promoter 3504..3554 /created_by="chgai" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /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 2023.0.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="KanR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (KanR)" /label="kan" misc_feature 3555..4364 /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.2 '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 4511..5099 /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" primer 5698..5718 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCDB-Cas9 - GWM13F" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13F (pCDB-Cas9 - GWM13F)" /label="GWM13F" ORIGIN 1 cgttgttcgt acacaaggat gtcgcttgtg gctttttcgc tcggctttcc aggagggcag 61 tccagatacc atgcggccgc cgtgtccatc actcgcgctg cagctgatgt gggcagatac 121 tgcactaatc tgccgcacga cgcgtcggcc tgagtcggcg gacaccggag gcttgctgga 181 gtactggcac aagtgtgcgt gtgattgcca gaaacagcaa cccactcact agatataccg 241 ctccaataat gggttcatgt gcaagctcac tcaataacga aacgcttcac tattgtgcac 301 gtgccggggt tgcttgggta tggcgtgcca tacgggttct gcacatgtca cccacgtggg 361 tgtggcacga gatatcgagg tggccatgta aaaggcattt gctagagggc agctcgtcca 421 tgccgctcga gcacaccgag tgcgcaccat aaatgcagca catgcctgtg cagcctatct 481 cctgtctctg ttacatccca ccacctcctc acatatcccc ttccaccctt aaaccgcaca 541 gcagacaaca atggccaagt gcgcatcgcg tgtcaatgct ggcgctgctg acgtccgctg 601 cctaaactgc atgaaagctc ctgcagtcgc aagtagtcct tttaacttgt tgtatcggaa 661 caaatggttg tagtctgtgc cgtgtgacga tcttgagccg cctggtcggc aggtccaatg 721 tcggcctgga ccctgcacac cctgcccacc ctgccgctgc tcacccgttc acgaccccac 781 aggtgctcgc agcagtaatt gtcgatagat agcaactgtg ttctgactat tctttgacct 841 tccgcatttg ggactgcggc ggcacaccta ctatccgcaa tgcacaccct ggtgctggta 901 ctgcctgcct ttccgcgcga gcccacacca tgcaggcgca tagtaatgga ctgtgattga 961 tttaggcaac tgtatgaaac ctcttcatcg tgccttggtt gcttgcgata ccctcacgtg 1021 agttcatatg accaacgctg gcacgacccc tgcaggatca ccagcgccgt tccggtgctc 1081 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg 1141 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc 1201 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg 1261 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg 1321 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg 1381 gccggcaact gcgtgcactt cgtggccgag gagcaggacg gcagcggcca gtgcacgaac 1441 tacgccctgc tgaagctggc gggcgacgtg gagagcaacc cgggcccggg ctcaacatga 1501 gaccgtcgac ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc 1561 ggtggctgag atcagccact tcttccccga taacggagac tggcacactg gccatatcgg 1621 tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacgggaga 1681 ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt 1741 caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt 1801 aaaccttaaa ctgcatttca ccagtccctg ttctcgtcag caaaagagcc gttcatttca 1861 ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc 1921 agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt gacatcatat 1981 atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag 2041 cacacctctt tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa 2101 atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc cacgcgttta 2161 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg 2221 gaagccatca cagacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc 2281 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac 2341 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc 2401 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata 2461 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc 2521 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc 2581 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat 2641 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc 2701 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc 2761 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt 2821 agcttcctta gctcctgaaa atctcgccgg atcctaactc aaaatccaca cattatacga 2881 gccggaagca taaagtgtaa agcctggggt gcctaatgcg gccgcggtct cagtatctga 2941 ctgcgcagtc aggcagagtc gttgttgggt gtatggttgc acaaaactag acctcgagct 3001 ggtcggttag aaagagatgc ttgtgcgact ggatgtaaca ggcacaatcc gaggaaaatg 3061 caacagaaac gttatgatgt aacatctgag cacagtcaat aaaattgcat gttgtatgcc 3121 gcacacaaaa gtgttgttaa acatgcgatg tggttgtcaa atcattattc gcacaaccat 3181 ggaccgaacc tctggcacgc tgcatacgct gtattcattc acatcatgtt ttctacttgt 3241 cacatgctct gcaccaagta actccgatta gttaggttcg cacatgtcat cggagtggac 3301 cacttcgaat catttggttc gaagataaac cggactataa gctaggtagt aactagtgcg 3361 actgtctaga atcgcgtatg cgagtgtatg agctcgatat cccctatagt gagtcgtatt 3421 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca 3481 ttgcacaaga taaaataata tcatcatgaa caataaaact gtctgcttac ataaacagta 3541 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca 3601 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg 3661 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca 3721 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat 3781 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca 3841 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg 3901 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta 3961 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata 4021 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag 4081 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg 4141 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg 4201 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg 4261 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata 4321 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt 4381 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa 4441 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca 4501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 4561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 4621 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 4681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 4741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 4801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 4861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 4921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 4981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 5041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 5101 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 5161 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 5221 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 5281 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 5341 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg 5401 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt 5461 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt 5521 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa 5581 acaacagata aaacgaaagg cccagacttc cgactgagcc tttcgtttta tttgatgcct 5641 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 5701 aaacgacggc cagtcttaag ctcgggccca cct //