LOCUS pGG-UmpRbcS-BleR-T2A-YFP-UmtRbcS 5031 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.p5-cgtpvik VERSION urn.local.jonasblomme.p5-cgtpvik 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..1482 /modified_by="jonasblomme" /label="T2A" Linker 1483..1512 /origin="C-linker-D (53bp linear)Fragment: from BsaI cut at 12^13 to BsaI cut at 40^41, 28bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" CDS 1519..2238 /ApEinfo_revcolor="#9eafd2" /ApEinfo_fwdcolor="#9eafd2" /modified_by="jonasblomme" /label="YFP" terminator 2243..2631 /regulatory_class="terminator" /note="Ulva RbcS terminator" /modified_by="jonasblomme" /label="UmtRbcS" Linker 2636..2684 /created_by="wacae" /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.4 '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(2721..2741) /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 2802..2852 /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 2853..3662 /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 3809..4397 /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 4996..5016 /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 gcagcggcga gggcagagga 1441 agtcttctaa catgcggtga cgtggaggag aatccgggcc ctggctccgt attcagtcga 1501 ctggtaccaa catcagctat ggtgagcaag ggcgaggagc tgttcacggg cgtggtgccc 1561 atcctggtgg agctggacgg cgacgtgaac ggccacaagt tcagcgtgtc cggcgagggc 1621 gagggcgacg ccacctacgg caagctgacc ctgaagttca tctgcacgac cggcaagctg 1681 cccgtgccct ggcccaccct ggtgaccacc ttcggctacg gcctgcagtg cttcgcccgc 1741 taccccgacc acatgaagca gcacgacttc ttcaagagcg ccatgccgga gggctacgtg 1801 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc ggaggtgaag 1861 ttcgagggcg acacgctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 1921 ggcaacatcc tgggccacaa gctggagtac aactacaaca gccacaacgt gtacatcatg 1981 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 2041 ggctcggtgc agctggcgga ccactaccag cagaacaccc ccatcggcga cggcccggtg 2101 ctgctgcccg acaaccacta cctgtcctac cagagcgccc tgtccaagga ccccaacgag 2161 aagcgcgacc acatggtgct gctggagttc gtgaccgccg ccggcatcac cctgggcatg 2221 gacgagctgt acaagtgact gcgcagtcag gcagagtcgt tgttgggtgt atggttgcac 2281 aaaactagac ctcgagctgg tcggttagaa agagatgctt gtgcgactgg atgtaacagg 2341 cacaatccga ggaaaatgca acagaaacgt tatgatgtaa catctgagca cagtcaataa 2401 aattgcatgt tgtatgccgc acacaaaagt gttgttaaac atgcgatgtg gttgtcaaat 2461 cattattcgc acaaccatgg accgaacctc tggcacgctg catacgctgt attcattcac 2521 atcatgtttt ctacttgtca catgctctgc accaagtaac tccgattagt taggttcgca 2581 catgtcatcg gagtggacca cttcgaatca tttggttcga agataaaccg gactataagc 2641 taggtagtaa ctagtgcgac tgtctagaat cgcgtatgcg agtgtatgag ctcgatatcc 2701 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc 2761 aaaatctctg atgttacatt gcacaagata aaataatatc atcatgaaca ataaaactgt 2821 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga 2881 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata 2941 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt 3001 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac 3061 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg 3121 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag 3181 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc 3241 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg 3301 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg 3361 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt 3421 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa 3481 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc 3541 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg 3601 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct 3661 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg 3721 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag 3781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 3841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 3901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 3961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 4021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 4081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 4141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 4201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 4261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 4321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 4381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 4441 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 4501 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 4561 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 4621 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 4681 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt 4741 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca 4801 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag 4861 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagacttccg actgagcctt 4921 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt 4981 cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggcccacc t //