LOCUS pGG-pUM140_0016-BleR-P2A-B-ccdB-CmR-G-UmtRbcS 7118 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.l-cn2ko84 VERSION urn.local.jonasblomme.l-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..2873 /label="P2A" GG complement(2879..2889) /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(2896..3201) /vntifkey=4 /label="ccdB" UTR complement(3202..3520) /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(3543..4202) /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(4256..4286) /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 4311..4321 /%_Identity=100 /Motif="GGTCTCAGTAT" /modified_by="mamee" /note="Derived using Geneious Prime 2023.0.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="G" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; G" /label="G" terminator 4330..4718 /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 4723..4771 /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(4808..4828) /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 4889..4939 /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 4940..5749 /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 5896..6484 /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 7083..7103 /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 agcggcgcta 2821 ctaatttttc tcttcttaag caagctggag atgttgaaga aaatcctgga cctggctcaa 2881 catgagaccg tcgacttata ttccccagaa catcaggtta atggcgtttt tgatgtcatt 2941 ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagactggca cactggccat 3001 atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt aaagttcacg 3061 ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc gtcgcccggg 3121 cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct ctcttttata 3181 ggtgtaaacc ttaaactgca tttcaccagt ccctgttctc gtcagcaaaa gagccgttca 3241 tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc cagcgttcgg 3301 cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag atattgacat 3361 catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa tacgctgctt 3421 catagcacac ctctttttga catacttcgg gtatacatat cagtatatat tcttataccg 3481 caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc ggatccacgc 3541 gtttacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg 3601 acatggaagc catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg 3661 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg 3721 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata 3781 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc 3841 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac 3901 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc 3961 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg 4021 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta 4081 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa 4141 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc 4201 attttagctt ccttagctcc tgaaaatctc gccggatcct aactcaaaat ccacacatta 4261 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgcggccgc ggtctcagta 4321 tctgactgcg cagtcaggca gagtcgttgt tgggtgtatg gttgcacaaa actagacctc 4381 gagctggtcg gttagaaaga gatgcttgtg cgactggatg taacaggcac aatccgagga 4441 aaatgcaaca gaaacgttat gatgtaacat ctgagcacag tcaataaaat tgcatgttgt 4501 atgccgcaca caaaagtgtt gttaaacatg cgatgtggtt gtcaaatcat tattcgcaca 4561 accatggacc gaacctctgg cacgctgcat acgctgtatt cattcacatc atgttttcta 4621 cttgtcacat gctctgcacc aagtaactcc gattagttag gttcgcacat gtcatcggag 4681 tggaccactt cgaatcattt ggttcgaaga taaaccggac tataagctag gtagtaacta 4741 gtgcgactgt ctagaatcgc gtatgcgagt gtatgagctc gatatcccct atagtgagtc 4801 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg 4861 ttacattgca caagataaaa taatatcatc atgaacaata aaactgtctg cttacataaa 4921 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa 4981 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc 5041 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca 5101 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac 5161 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt 5221 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc 5281 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt 5341 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat 5401 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga 5461 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca 5521 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga 5581 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct 5641 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc 5701 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt 5761 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg 5821 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa 5881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 5941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 6001 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 6061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 6121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 6181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 6241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 6301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 6361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 6421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 6481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 6541 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 6601 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 6661 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 6721 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg 6781 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc 6841 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt 6901 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc 6961 gacaaacaac agataaaacg aaaggcccag acttccgact gagcctttcg ttttatttga 7021 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg 7081 ttgtaaaacg acggccagtc ttaagctcgg gcccacct //