LOCUS pGG-UmpRbcS-BleR-E2A-C-ccdB-CmR-E-UmtRbcS 5720 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.q-cn2ko85 VERSION urn.local.jonasblomme.q-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..1488 /label="E2A" GG complement(1489..1499) /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(1506..1811) /vntifkey=4 /label="ccdB" UTR complement(1812..2130) /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(2153..2812) /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(2866..2896) /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 2921..2931 /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 2932..3320 /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 3325..3373 /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(3410..3430) /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 3491..3541 /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 3542..4351 /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 4498..5086 /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 5685..5705 /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 gcagcggcca gtgcacgaac 1441 tacgccctgc tgaagctggc gggcgacgtg gagagcaacc cgggcccggg cttgagaccg 1501 tcgacttata ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg 1561 ctgagatcag ccacttcttc cccgataacg gagactggca cactggccat atcggtggtc 1621 atcatgcgcc agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta 1681 tctgacagca gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata 1741 atatcactct gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc 1801 ttaaactgca tttcaccagt ccctgttctc gtcagcaaaa gagccgttca tttcaataaa 1861 ccgggcgacc tcagccatcc cttcctgatt ttccgctttc cagcgttcgg cacgcagacg 1921 acgggcttca ttctgcatgg ttgtgcttac cagaccggag atattgacat catatatgcc 1981 ttgagcaact gatagctgtc gctgtcaact gtcactgtaa tacgctgctt catagcacac 2041 ctctttttga catacttcgg gtatacatat cagtatatat tcttataccg caaaaatcag 2101 cgcgcaaata cgcatactgt tatctggctt ttagtaagcc ggatccacgc gtttacgccc 2161 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc 2221 catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg 2281 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta 2341 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa 2401 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt 2461 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt 2521 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt 2581 tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg 2641 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct 2701 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac 2761 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt 2821 ccttagctcc tgaaaatctc gccggatcct aactcaaaat ccacacatta tacgagccgg 2881 aagcataaag tgtaaagcct ggggtgccta atgcggccgc ggtctcactg cgcagtcagg 2941 cagagtcgtt gttgggtgta tggttgcaca aaactagacc tcgagctggt cggttagaaa 3001 gagatgcttg tgcgactgga tgtaacaggc acaatccgag gaaaatgcaa cagaaacgtt 3061 atgatgtaac atctgagcac agtcaataaa attgcatgtt gtatgccgca cacaaaagtg 3121 ttgttaaaca tgcgatgtgg ttgtcaaatc attattcgca caaccatgga ccgaacctct 3181 ggcacgctgc atacgctgta ttcattcaca tcatgttttc tacttgtcac atgctctgca 3241 ccaagtaact ccgattagtt aggttcgcac atgtcatcgg agtggaccac ttcgaatcat 3301 ttggttcgaa gataaaccgg actataagct aggtagtaac tagtgcgact gtctagaatc 3361 gcgtatgcga gtgtatgagc tcgatatccc ctatagtgag tcgtattaca tggtcatagc 3421 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa 3481 aataatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt 3541 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga 3601 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg 3661 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc 3721 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc 3781 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc 3841 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga 3901 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa 3961 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga 4021 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat 4081 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga 4141 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat 4201 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc 4261 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca 4321 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg 4381 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt 4441 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 4621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt 5161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 5221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg 5281 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 5341 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga 5401 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg 5461 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa 5521 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa 5581 cgaaaggccc agacttccga ctgagccttt cgttttattt gatgcctggc agttccctac 5641 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag 5701 tcttaagctc gggcccacct //