LOCUS pGG-pUM140_0016-BleR-F2A-B-ccdB-CmR-G-UmtRbcS 7127 bp DNA circular UNA 11-APR-2023 DEFINITION . ACCESSION urn.local.jonasblomme.j-cn2ko84 VERSION urn.local.jonasblomme.j-cn2ko84 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" GG complement(2888..2898) /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(2905..3210) /vntifkey=4 /label="ccdB" UTR complement(3211..3529) /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(3552..4211) /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(4265..4295) /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 4320..4330 /%_Identity=100 /Motif="GGTCTCAGTAT" /modified_by="mamee" /note="Derived using Geneious Prime 2023.0.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="G" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; G" /label="G" terminator 4339..4727 /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 4732..4780 /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(4817..4837) /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 4898..4948 /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 4949..5758 /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 5905..6493 /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 7092..7112 /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 ccggctcaac atgagaccgt cgacttatat tccccagaac atcaggttaa tggcgttttt 2941 gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg agactggcac 3001 actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca ccaccgggta 3061 aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga tcaccatccg 3121 tcgcccgggc gtgtcaataa tatcactctg tacatccaca aacagacgat aacggctctc 3181 tcttttatag gtgtaaacct taaactgcat ttcaccagtc cctgttctcg tcagcaaaag 3241 agccgttcat ttcaataaac cgggcgacct cagccatccc ttcctgattt tccgctttcc 3301 agcgttcggc acgcagacga cgggcttcat tctgcatggt tgtgcttacc agaccggaga 3361 tattgacatc atatatgcct tgagcaactg atagctgtcg ctgtcaactg tcactgtaat 3421 acgctgcttc atagcacacc tctttttgac atacttcggg tatacatatc agtatatatt 3481 cttataccgc aaaaatcagc gcgcaaatac gcatactgtt atctggcttt tagtaagccg 3541 gatccacgcg tttacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc 3601 attctgccga catggaagcc atcacagacg gcatgatgaa cctgaatcgc cagcggcatc 3661 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg 3721 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg 3781 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc 3841 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc 3901 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat 3961 atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg 4021 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa 4081 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat 4141 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt 4201 tttttctcca ttttagcttc cttagctcct gaaaatctcg ccggatccta actcaaaatc 4261 cacacattat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgcggccgcg 4321 gtctcagtat ctgactgcgc agtcaggcag agtcgttgtt gggtgtatgg ttgcacaaaa 4381 ctagacctcg agctggtcgg ttagaaagag atgcttgtgc gactggatgt aacaggcaca 4441 atccgaggaa aatgcaacag aaacgttatg atgtaacatc tgagcacagt caataaaatt 4501 gcatgttgta tgccgcacac aaaagtgttg ttaaacatgc gatgtggttg tcaaatcatt 4561 attcgcacaa ccatggaccg aacctctggc acgctgcata cgctgtattc attcacatca 4621 tgttttctac ttgtcacatg ctctgcacca agtaactccg attagttagg ttcgcacatg 4681 tcatcggagt ggaccacttc gaatcatttg gttcgaagat aaaccggact ataagctagg 4741 tagtaactag tgcgactgtc tagaatcgcg tatgcgagtg tatgagctcg atatccccta 4801 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa 4861 tctctgatgt tacattgcac aagataaaat aatatcatca tgaacaataa aactgtctgc 4921 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc 4981 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt 5041 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt 5101 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa 5161 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga 5221 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata 5281 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc 5341 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca 5401 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg 5461 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt 5521 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg 5581 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg 5641 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat 5701 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc 5761 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc 5821 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc 5881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 5941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 6001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 6061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 6121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 6181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 6241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 6301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 6361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 6421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 6481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 6541 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 6601 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 6661 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 6721 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 6781 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg 6841 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct 6901 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga 6961 gcgttcaccg acaaacaaca gataaaacga aaggcccaga cttccgactg agcctttcgt 7021 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca 7081 gtcacgacgt tgtaaaacga cggccagtct taagctcggg cccacct //