LOCUS pGG-pUM140_0016-BleR-T2A-C-ccdB-CmR-E-UmtRbcS 7103 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.o-cn2ko84 VERSION urn.local.jonasblomme.o-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..2871 /label="T2A" GG complement(2872..2882) /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(2889..3194) /vntifkey=4 /label="ccdB" UTR complement(3195..3513) /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(3536..4195) /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(4249..4279) /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 4304..4314 /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 4315..4703 /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 4708..4756 /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(4793..4813) /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 4874..4924 /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 4925..5734 /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 5881..6469 /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 7068..7088 /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 agcggcgagg 2821 gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tccgggccct gggcttgaga 2881 ccgtcgactt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg 2941 tggctgagat cagccacttc ttccccgata acggagactg gcacactggc catatcggtg 3001 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact 3061 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca 3121 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa 3181 accttaaact gcatttcacc agtccctgtt ctcgtcagca aaagagccgt tcatttcaat 3241 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag 3301 acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga catcatatat 3361 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca 3421 cacctctttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat 3481 cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcca cgcgtttacg 3541 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga 3601 agccatcaca gacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt 3661 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt 3721 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa 3781 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata 3841 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag 3901 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt 3961 ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa 4021 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca 4081 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt 4141 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag 4201 cttccttagc tcctgaaaat ctcgccggat cctaactcaa aatccacaca ttatacgagc 4261 cggaagcata aagtgtaaag cctggggtgc ctaatgcggc cgcggtctca ctgcgcagtc 4321 aggcagagtc gttgttgggt gtatggttgc acaaaactag acctcgagct ggtcggttag 4381 aaagagatgc ttgtgcgact ggatgtaaca ggcacaatcc gaggaaaatg caacagaaac 4441 gttatgatgt aacatctgag cacagtcaat aaaattgcat gttgtatgcc gcacacaaaa 4501 gtgttgttaa acatgcgatg tggttgtcaa atcattattc gcacaaccat ggaccgaacc 4561 tctggcacgc tgcatacgct gtattcattc acatcatgtt ttctacttgt cacatgctct 4621 gcaccaagta actccgatta gttaggttcg cacatgtcat cggagtggac cacttcgaat 4681 catttggttc gaagataaac cggactataa gctaggtagt aactagtgcg actgtctaga 4741 atcgcgtatg cgagtgtatg agctcgatat cccctatagt gagtcgtatt acatggtcat 4801 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga 4861 taaaataata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg 4921 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc 4981 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta 5041 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt 5101 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct 5161 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat 5221 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt 5281 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt 5341 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt 5401 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga 5461 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact 5521 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg 5581 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc 5641 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt 5701 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca 5761 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa 5821 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 5881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 5941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6001 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 6061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6541 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 6601 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 6661 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 6721 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga 6781 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc 6841 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc 6901 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata 6961 aaacgaaagg cccagacttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc 7021 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc 7081 cagtcttaag ctcgggccca cct //