LOCUS pGG-UmpRbcS-BleR-P2A-B-ccdB-CmR-G-UmtRbcS 5730 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.t-cn2ko85 VERSION urn.local.jonasblomme.t-cn2ko85 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..1485 /label="P2A" GG complement(1491..1501) /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(1508..1813) /vntifkey=4 /label="ccdB" UTR complement(1814..2132) /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(2155..2814) /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(2868..2898) /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 2923..2933 /%_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 2942..3330 /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 3335..3383 /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(3420..3440) /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 3501..3551 /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 3552..4361 /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 4508..5096 /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 5695..5715 /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 gcagcggcgc tactaatttt 1441 tctcttctta agcaagctgg agatgttgaa gaaaatcctg gacctggctc aacatgagac 1501 cgtcgactta tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt 1561 ggctgagatc agccacttct tccccgataa cggagactgg cacactggcc atatcggtgg 1621 tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt 1681 tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa 1741 taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa 1801 ccttaaactg catttcacca gtccctgttc tcgtcagcaa aagagccgtt catttcaata 1861 aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga 1921 cgacgggctt cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg 1981 ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcac 2041 acctcttttt gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc 2101 agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatccac gcgtttacgc 2161 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa 2221 gccatcacag acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg 2281 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt 2341 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat 2401 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat 2461 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt 2521 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc 2581 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa 2641 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag 2701 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt 2761 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc 2821 ttccttagct cctgaaaatc tcgccggatc ctaactcaaa atccacacat tatacgagcc 2881 ggaagcataa agtgtaaagc ctggggtgcc taatgcggcc gcggtctcag tatctgactg 2941 cgcagtcagg cagagtcgtt gttgggtgta tggttgcaca aaactagacc tcgagctggt 3001 cggttagaaa gagatgcttg tgcgactgga tgtaacaggc acaatccgag gaaaatgcaa 3061 cagaaacgtt atgatgtaac atctgagcac agtcaataaa attgcatgtt gtatgccgca 3121 cacaaaagtg ttgttaaaca tgcgatgtgg ttgtcaaatc attattcgca caaccatgga 3181 ccgaacctct ggcacgctgc atacgctgta ttcattcaca tcatgttttc tacttgtcac 3241 atgctctgca ccaagtaact ccgattagtt aggttcgcac atgtcatcgg agtggaccac 3301 ttcgaatcat ttggttcgaa gataaaccgg actataagct aggtagtaac tagtgcgact 3361 gtctagaatc gcgtatgcga gtgtatgagc tcgatatccc ctatagtgag tcgtattaca 3421 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg 3481 cacaagataa aataatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata 3541 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca 3601 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga 3661 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag 3721 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta 3781 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca 3841 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa 3901 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt 3961 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg 4021 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct 4081 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt 4141 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac 4201 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt 4261 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga 4321 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt 4381 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat 4441 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 4621 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 5101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 5161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 5221 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 5281 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 5341 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta 5401 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt 5461 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac 5521 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca 5581 acagataaaa cgaaaggccc agacttccga ctgagccttt cgttttattt gatgcctggc 5641 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa 5701 cgacggccag tcttaagctc gggcccacct //