LOCUS pGG-pUM140_0016-BleR-F2A-mCherry-UmtRbcS 6422 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.p7-cgtpvik VERSION urn.local.jonasblomme.p7-cgtpvik 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..2882 /label="F2A" Linker 2883..2912 /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 2919..3626 /SerialCloner_Color="&hFF0000" /SerialCloner_Show="True" /SerialCloner_Protect="True" /SerialCloner_Arrow="True" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="mCherry" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; mCherry" /modified_by="jonasblomme" /label="mCherry" terminator 3634..4022 /regulatory_class="terminator" /note="Ulva RbcS terminator" /label="UmtRbcS" Linker 4027..4075 /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(4112..4132) /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 4193..4243 /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 4244..5053 /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 5200..5788 /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 6387..6407 /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 agcggcgtga 2821 agcagaccct gaacttcgac ctgctgaagc tggcgggcga cgtggagagc aacccgggcc 2881 ccggctccgt attcagtcga ctggtaccaa catcagctat ggtgtccaag ggcgaggagg 2941 acaacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggcagcgtga 3001 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 3061 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgagcc 3121 cccagttcat gtacggcagc aaggcctacg tgaagcaccc cgccgacatc cccgactacc 3181 tgaagctgag cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 3241 gcgtggtgac cgtgacccag gacagcagcc tccaggacgg cgagttcatc tacaaggtga 3301 agctgcgcgg caccaacttc cccagcgacg gccccgtgat gcagaagaag accatgggct 3361 gggaggccag cagcgagcgc atgtaccccg aggacggcgc cctgaagggc gagatcaagc 3421 agcgcctgaa gctgaaggac ggcggccact acgacgccga ggtgaagacc acctacaagg 3481 ccaagaagcc cgtgcagctg cccggcgcct acaacgtgaa catcaagctg gacatcacca 3541 gccacaacga ggactacacc atcgtggagc agtacgagcg cgctgagggc cgccacagca 3601 ccggcggcat ggacgagctg tacaagtaac tgcgcagtca ggcagagtcg ttgttgggtg 3661 tatggttgca caaaactaga cctcgagctg gtcggttaga aagagatgct tgtgcgactg 3721 gatgtaacag gcacaatccg aggaaaatgc aacagaaacg ttatgatgta acatctgagc 3781 acagtcaata aaattgcatg ttgtatgccg cacacaaaag tgttgttaaa catgcgatgt 3841 ggttgtcaaa tcattattcg cacaaccatg gaccgaacct ctggcacgct gcatacgctg 3901 tattcattca catcatgttt tctacttgtc acatgctctg caccaagtaa ctccgattag 3961 ttaggttcgc acatgtcatc ggagtggacc acttcgaatc atttggttcg aagataaacc 4021 ggactataag ctaggtagta actagtgcga ctgtctagaa tcgcgtatgc gagtgtatga 4081 gctcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg 4141 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat catcatgaac 4201 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg 4261 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg 4321 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga 4381 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga 4441 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat 4501 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca 4561 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct 4621 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg 4681 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga 4741 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt 4801 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga 4861 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga 4921 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt 4981 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga 5041 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac 5101 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca 5161 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 5221 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 5281 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 5341 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 5401 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 5461 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 5521 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 5581 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 5641 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 5701 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 5761 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 5821 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 5881 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 5941 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 6001 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 6061 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa 6121 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt 6181 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt 6241 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagacttcc 6301 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc 6361 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccac 6421 ct //