LOCUS pGG-UmpRbcS-BleR-P2A-C-ccdB-CmR-E-UmtRbcS 5717 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.u-cn2ko85 VERSION urn.local.jonasblomme.u-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(1486..1496) /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(1503..1808) /vntifkey=4 /label="ccdB" UTR complement(1809..2127) /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(2150..2809) /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(2863..2893) /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 2918..2928 /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 2929..3317 /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 3322..3370 /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(3407..3427) /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 3488..3538 /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 3539..4348 /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 4495..5083 /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 5682..5702 /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 gacctggctt gagaccgtcg 1501 acttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg 1561 agatcagcca cttcttcccc gataacggag actggcacac tggccatatc ggtggtcatc 1621 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct 1681 gacagcagac gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata 1741 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta 1801 aactgcattt caccagtccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg 1861 ggcgacctca gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg 1921 ggcttcattc tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg 1981 agcaactgat agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcacacctc 2041 tttttgacat acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc 2101 gcaaatacgc atactgttat ctggctttta gtaagccgga tccacgcgtt tacgccccgc 2161 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat 2221 cacagacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat 2281 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat 2341 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc 2401 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta 2461 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct 2521 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca 2581 ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg 2641 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa 2701 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat 2761 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct 2821 tagctcctga aaatctcgcc ggatcctaac tcaaaatcca cacattatac gagccggaag 2881 cataaagtgt aaagcctggg gtgcctaatg cggccgcggt ctcactgcgc agtcaggcag 2941 agtcgttgtt gggtgtatgg ttgcacaaaa ctagacctcg agctggtcgg ttagaaagag 3001 atgcttgtgc gactggatgt aacaggcaca atccgaggaa aatgcaacag aaacgttatg 3061 atgtaacatc tgagcacagt caataaaatt gcatgttgta tgccgcacac aaaagtgttg 3121 ttaaacatgc gatgtggttg tcaaatcatt attcgcacaa ccatggaccg aacctctggc 3181 acgctgcata cgctgtattc attcacatca tgttttctac ttgtcacatg ctctgcacca 3241 agtaactccg attagttagg ttcgcacatg tcatcggagt ggaccacttc gaatcatttg 3301 gttcgaagat aaaccggact ataagctagg tagtaactag tgcgactgtc tagaatcgcg 3361 tatgcgagtg tatgagctcg atatccccta tagtgagtcg tattacatgg tcatagctgt 3421 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaat 3481 aatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat 3541 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt 3601 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt 3661 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa 3721 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac 3781 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg 3841 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc 3901 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag 3961 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc 4021 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca 4081 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa 4141 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc 4201 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt 4261 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt 4321 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca 4381 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag 4441 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 4621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 5101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 5161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 5221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa 5281 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga 5341 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt 5401 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca 5461 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc 5521 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga 5581 aaggcccaga cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct 5641 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct 5701 taagctcggg cccacct //