LOCUS       pGG-pUM140_0016-BleR-F2A-CreYFP-2xSV40-UmtRbcS        6485 bp    DNA     circular UNA 12-APR-2023
DEFINITION  .
ACCESSION   urn.local.jonasblomme.ix-cgtpsou
VERSION     urn.local.jonasblomme.ix-cgtpsou
KEYWORDS    .
SOURCE      
  ORGANISM  .
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..3635
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /modified_by="jonasblomme"
                     /label="YFP"
     NLS             3636..3689
     terminator      3697..4085
                     /regulatory_class="terminator"
                     /note="Ulva RbcS terminator"
                     /modified_by="jonasblomme"
                     /label="UmtRbcS"
     Linker          4090..4138
                     /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; <a
                     href=""urn:local:thjac:1h-c4nnd54"">Linker II</a>"
                     /label="Linker II"
     primer          complement(4175..4195)
                     /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        4256..4306
                     /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 (<a
                     href=""urn:local:chgai:ll-ch8iv1b"">pCG034_pKan_Kan-CBE</a
                     >)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lo-ch8iv1b"">pCG040_ABE-Nuc-NLS_Kan
                     -reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lr-ch8iv1b"">pCG043_Kan-ABE-Dest_co
                     ntrol_ccdB</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lu-ch8iv1c"">pCG046_ABE-LbCas12a-NL
                     S_Kan_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:kx-ch8iv19"">pCG005_pGGD-pKan-KanR-
                     E</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ky-ch8iv19"">pCG006_pGGA-Kan_promot
                     er-B</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lm-ch8iv1b"">pCG035_pKan_Kan</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ln-ch8iv1b"">pCG039_ABE-N7-NLS_Kan-
                     reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lp-ch8iv1b"">pCG041_ABE-SV40-NLS_Ka
                     n-reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lw-ch8iv1c"">pCG054_Kan_CBE_control
                     </a>)"
                     /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 (<a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>)"
                     /label="kan"
     misc_feature    4307..5116
                     /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; <a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>"
                     /label="KanR"
                     /note="Geneious type: Bacterial Selection"
     rep_origin      5263..5851
                     /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 (<a
                     href=""urn:local:.:8d-dk4mzop"">high-copy-number
                     ColE1/pMB1/pBR322/pUC origin of  replication</a>)"
                     /label="ColE1/pMB1/pBR322/pUC Ori"
     primer          6450..6470
                     /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 ggtgagcaag ggcgaggagc
     2941 tgttcacggg cgtggtgccc atcctggtgg agctggacgg cgacgtgaac ggccacaagt
     3001 tcagcgtgtc cggcgagggc gagggcgacg ccacctacgg caagctgacc ctgaagttca
     3061 tctgcacgac cggcaagctg cccgtgccct ggcccaccct ggtgaccacc ttcggctacg
     3121 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagagcg
     3181 ccatgccgga gggctacgtg caggagcgca ccatcttctt caaggacgac ggcaactaca
     3241 agacccgcgc ggaggtgaag ttcgagggcg acacgctggt gaaccgcatc gagctgaagg
     3301 gcatcgactt caaggaggac ggcaacatcc tgggccacaa gctggagtac aactacaaca
     3361 gccacaacgt gtacatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3421 tccgccacaa catcgaggac ggctcggtgc agctggcgga ccactaccag cagaacaccc
     3481 ccatcggcga cggcccggtg ctgctgcccg acaaccacta cctgtcctac cagagcgccc
     3541 tgtccaagga ccccaacgag aagcgcgacc acatggtgct gctggagttc gtgaccgccg
     3601 ccggcatcac cctgggcatg gacgagctgt acaaggaccc gaagaagaag cgcaaggtgg
     3661 acccgaagaa gaagcgcaag gtggacccct gactgcgcag tcaggcagag tcgttgttgg
     3721 gtgtatggtt gcacaaaact agacctcgag ctggtcggtt agaaagagat gcttgtgcga
     3781 ctggatgtaa caggcacaat ccgaggaaaa tgcaacagaa acgttatgat gtaacatctg
     3841 agcacagtca ataaaattgc atgttgtatg ccgcacacaa aagtgttgtt aaacatgcga
     3901 tgtggttgtc aaatcattat tcgcacaacc atggaccgaa cctctggcac gctgcatacg
     3961 ctgtattcat tcacatcatg ttttctactt gtcacatgct ctgcaccaag taactccgat
     4021 tagttaggtt cgcacatgtc atcggagtgg accacttcga atcatttggt tcgaagataa
     4081 accggactat aagctaggta gtaactagtg cgactgtcta gaatcgcgta tgcgagtgta
     4141 tgagctcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     4201 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaataa tatcatcatg
     4261 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     4321 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     4381 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     4441 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     4501 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     4561 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     4621 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     4681 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     4741 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     4801 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     4861 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     4921 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     4981 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     5041 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     5101 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     5161 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     5221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5881 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5941 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6001 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6061 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6121 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     6181 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     6241 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     6301 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagact
     6361 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     6421 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     6481 cacct
//
