LOCUS pGG-pUM140_0016-BleR-E2A-C-ccdB-CmR-E-UmtRbcS 7108 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.i-cn2ko84 VERSION urn.local.jonasblomme.i-cn2ko84 KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/. FEATURES Location/Qualifiers promoter 1..1934 /modified_by="jonasblomme" /label="pUM140_0016" CDS join(1939..1946,2444..2807) /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 1947..2443 /label="RbcsI" CDS 2808..2876 /label="E2A" GG complement(2877..2887) /created_by="thjac" /modified_by="thjac" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="C" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; C" /label="C" CDS complement(2894..3199) /vntifkey=4 /label="ccdB" UTR complement(3200..3518) /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(3541..4200) /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(4254..4284) /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 4309..4319 /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="E" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; E" /label="E" terminator 4320..4708 /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 4713..4761 /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(4798..4818) /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 4879..4929 /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 4930..5739 /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 5886..6474 /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 7073..7093 /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 tgccgaattc gcagtgaacc cgcatggcag ggtggtcaca ggtgtccttt ggctgcacag 61 taacatgacc gtcatgcatg ggacagaggt tgtgagcagc atgcgtcgat gcatacatgc 121 gcatgtagca tgcagcagtg agctccagag tggaaagctg gcccgctgtc gtgtgtccgg 181 accgacaggc caggctctgg actgggccat agtcagcagg gcagaggtac tatgtctagc 241 gccttcctgg gtgtgcagct ttgccttgcc atgcttggac atgaatgagt ggggcgccag 301 gcacttacaa gagccacctc gtggtgtggg catatggcag aggggcaccc atcagtcgtg 361 tgccattcag accgtgacga agcaatcaat gactgtgctt gtccgggaga aagtctgttg 421 tgaaatttgc tgtgatctag gggccaatgt ggcagtcagg aatggctaga gatgctgcta 481 gtgtcgtgca tacctggtgt actgccaaca gcctcacctg accatggtgc ttgtgtttaa 541 agcaagcatg ccacaagtcg gcatgtaggc atacggcaag agctacacag ttggctgttg 601 acctgtggat gcgtgaaaca gaagtttgaa tcgtgccctt tcgcgttgat gcaccctgtg 661 tgggagccag gctggtgtgt gtggaagcca ggctggtggc acacctgtgt gacctggagg 721 tggacgaccg ggtggcgagg caagtagcag tttcatcaaa gaaacacaag ctgatgcatc 781 cgcgatagta gcacgattct aatgtgtgca cgcaaacgca tcggcccaac tggagcaggg 841 gatgcgcagc agcggccgct ggttccacga actcgaagcg caaagagcag tccgctgtga 901 ccagcattcc gctagtgtca gggacgatat gaatacgggt tgggcacatt atgtttgggt 961 cgaggtcgcc atggcggacg tgcaagcggc tgtcgatcac gcctatcgga gatgcccagc 1021 gcctgcatcg atatgcaacc agctctgcac cagcgcagca atacgacagc tgcagccgat 1081 ggcctcctag acactgtgca acaacaaaaa acaattggac cgtgacggca caggtgtcga 1141 acacaggggc tgagagagga ctgcctgtgc ttgtgctcca gtgctcatac gccatactga 1201 tacgcctcca ccgatggcct cactgcctgc ggacactggc agacgctgtc gtttggatgt 1261 cgcatcgcca tcttcagtga tgttgtggcg ttcggcagga tgtacgcatg agcgattctt 1321 ctaaacgagc agtggtggca catcagcggc tccctcaaac cacaatgccg gcttggaagg 1381 tagagcatct gctcggcaag ccgacataga cgatacagct agcacagccg gcttgcagct 1441 catcaatcag tacacatacc gtacgtgtct gaagtatgcc ttgttgatat gaacactgta 1501 ccgtggggcc agacgacacg gtctgaagca ccgcctctgc agcatctcgc acgattgatg 1561 cgcatcaatt gcttgcaatt cgatttgcat ttggcgcagt tgtctgcaca atctttgcca 1621 acgatcgcat gatccgattc aatgatccga agcatgatca atacaccgca ttagcagtgc 1681 atgcgcgatt gggacacctc tgggggggga tctgatgcac gttcgacacg ggtgccgaac 1741 ctgcgagaag gctcactaca cgcgtgcgga cgcatgcatg cagtgccaaa gatctcgttt 1801 gaactgcgcg ccagcacatt gcagtgcacg ccccgtgcca ggcgaaaacg tatttaaccc 1861 cgaagccagc atactttctc taccagttca tctcttgttg atctacctcc accacagtct 1921 atcagtgatt aacaaacaat ggccaagtgc gcatcgcgtg tcaatgctgg cgctgctgac 1981 gtccgctgcc taaactgcat gaaagctcct gcagtcgcaa gtagtccttt taacttgttg 2041 tatcggaaca aatggttgta gtctgtgccg tgtgacgatc ttgagccgcc tggtcggcag 2101 gtccaatgtc ggcctggacc ctgcacaccc tgcccaccct gccgctgctc acccgttcac 2161 gaccccacag gtgctcgcag cagtaattgt cgatagatag caactgtgtt ctgactattc 2221 tttgaccttc cgcatttggg actgcggcgg cacacctact atccgcaatg cacaccctgg 2281 tgctggtact gcctgccttt ccgcgcgagc ccacaccatg caggcgcata gtaatggact 2341 gtgattgatt taggcaactg tatgaaacct cttcatcgtg ccttggttgc ttgcgatacc 2401 ctcacgtgag ttcatatgac caacgctggc acgacccctg caggatcacc agcgccgttc 2461 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt 2521 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt 2581 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc 2641 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg 2701 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc 2761 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggacggc agcggccagt 2821 gcacgaacta cgccctgctg aagctggcgg gcgacgtgga gagcaacccg ggcccgggct 2881 tgagaccgtc gacttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt 2941 cgcggtggct gagatcagcc acttcttccc cgataacgga gactggcaca ctggccatat 3001 cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg 3061 agactttatc tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg 3121 tgtcaataat atcactctgt acatccacaa acagacgata acggctctct cttttatagg 3181 tgtaaacctt aaactgcatt tcaccagtcc ctgttctcgt cagcaaaaga gccgttcatt 3241 tcaataaacc gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca 3301 cgcagacgac gggcttcatt ctgcatggtt gtgcttacca gaccggagat attgacatca 3361 tatatgcctt gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca 3421 tagcacacct ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca 3481 aaaatcagcg cgcaaatacg catactgtta tctggctttt agtaagccgg atccacgcgt 3541 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac 3601 atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc 3661 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc 3721 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt 3781 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga 3841 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt 3901 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc 3961 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg 4021 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat 4081 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg 4141 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat 4201 tttagcttcc ttagctcctg aaaatctcgc cggatcctaa ctcaaaatcc acacattata 4261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gcggccgcgg tctcactgcg 4321 cagtcaggca gagtcgttgt tgggtgtatg gttgcacaaa actagacctc gagctggtcg 4381 gttagaaaga gatgcttgtg cgactggatg taacaggcac aatccgagga aaatgcaaca 4441 gaaacgttat gatgtaacat ctgagcacag tcaataaaat tgcatgttgt atgccgcaca 4501 caaaagtgtt gttaaacatg cgatgtggtt gtcaaatcat tattcgcaca accatggacc 4561 gaacctctgg cacgctgcat acgctgtatt cattcacatc atgttttcta cttgtcacat 4621 gctctgcacc aagtaactcc gattagttag gttcgcacat gtcatcggag tggaccactt 4681 cgaatcattt ggttcgaaga taaaccggac tataagctag gtagtaacta gtgcgactgt 4741 ctagaatcgc gtatgcgagt gtatgagctc gatatcccct atagtgagtc gtattacatg 4801 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca 4861 caagataaaa taatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca 4921 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg 4981 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca 5041 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt 5101 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg 5161 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact 5221 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat 5281 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt 5341 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt 5401 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg 5461 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc 5521 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga 5581 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt 5641 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat 5701 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg 5761 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc 5821 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 5881 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 5941 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 6001 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 6061 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 6121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 6181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 6241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 6301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 6361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 6421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 6481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 6541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 6601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 6661 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 6721 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc 6781 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt 6841 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa 6901 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac 6961 agataaaacg aaaggcccag acttccgact gagcctttcg ttttatttga tgcctggcag 7021 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg 7081 acggccagtc ttaagctcgg gcccacct //