LOCUS pGG-pUM140_0016-BleR-T2A-B-ccdB-CmR-G-UmtRbcS 7116 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.n-cn2ko84 VERSION urn.local.jonasblomme.n-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(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="B" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; B" /label="B" 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 /%_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 4328..4716 /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 4721..4769 /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(4806..4826) /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 4887..4937 /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 4938..5747 /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 5894..6482 /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 7081..7101 /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 gggctcaaca 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 tctcagtatc 4321 tgactgcgca gtcaggcaga gtcgttgttg ggtgtatggt tgcacaaaac tagacctcga 4381 gctggtcggt tagaaagaga tgcttgtgcg actggatgta acaggcacaa tccgaggaaa 4441 atgcaacaga aacgttatga tgtaacatct gagcacagtc aataaaattg catgttgtat 4501 gccgcacaca aaagtgttgt taaacatgcg atgtggttgt caaatcatta ttcgcacaac 4561 catggaccga acctctggca cgctgcatac gctgtattca ttcacatcat gttttctact 4621 tgtcacatgc tctgcaccaa gtaactccga ttagttaggt tcgcacatgt catcggagtg 4681 gaccacttcg aatcatttgg ttcgaagata aaccggacta taagctaggt agtaactagt 4741 gcgactgtct agaatcgcgt atgcgagtgt atgagctcga tatcccctat agtgagtcgt 4801 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt 4861 acattgcaca agataaaata atatcatcat gaacaataaa actgtctgct tacataaaca 4921 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt 4981 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag 5041 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg 5101 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg 5161 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac 5221 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag 5281 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt 5341 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga 5401 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac 5461 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg 5521 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg 5581 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg 5641 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg 5701 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta 5761 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac 5821 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga 5881 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 5941 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 6001 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 6061 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 6121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 6181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 6241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 6301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 6361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 6421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 6481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 6541 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 6601 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 6661 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct 6721 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta 6781 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg 6841 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg 6901 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga 6961 caaacaacag ataaaacgaa aggcccagac ttccgactga gcctttcgtt ttatttgatg 7021 cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt 7081 gtaaaacgac ggccagtctt aagctcgggc ccacct //