LOCUS       sequence-169707-        3286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate Level 0 cassette encoding the U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3286)
  AUTHORS   Hopes A, Nekrasov V, Kamoun S, Mock T
  TITLE     Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira
            pseudonana.
  JOURNAL   Plant Methods. 2016 Nov 24;12:49. doi: 10.1186/s13007-016-0148-0. 
            eCollection 2016.
  PUBMED    27904648
REFERENCE   2  (bases 1 to 3286)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Methods."; date: "2016-11-24"; volume: "12:49. doi"; pages: " 
            10.1186/s13007-016-0148-0. eCollection 2016"
COMMENT     Sequence Label: pCR8/GW:U6
FEATURES             Location/Qualifiers
     source          1..3286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(143..162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(744..1535)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     1966..1982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1987..2006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1987..2005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2023..2122
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2619..2718)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2734..2751)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2734..2750)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2743..2765)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2814..2900
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2992..3019
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3178..3195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
       61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat
      721 tttggtcatg ggtggctcga gggttatttg ccgactacct tggtgatctc gcctttcacg
      781 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
      841 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
      901 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
      961 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     1021 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     1081 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     1141 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     1201 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     1261 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     1321 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     1381 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     1441 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     1501 tgagtcgata cttcggcgat caccgcttcc ctcataatgt ttaactttgt tttagggcga
     1561 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     1621 cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa
     1681 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     1741 ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac
     1801 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggtagcagc
     1861 gaagtcgagg catttctgtc ctggctggtc tagaggaggc atgatgatat atttttatct
     1921 tgtgcaatgt aacatcagag attttgagac acgggccaga gctgccagga aacagctatg
     1981 accatgtaat acgactcact ataggggata tcagctggat ggcaaataat gattttattt
     2041 tgactgatag tgacctgttc gttgcaacaa attgatgagc aattattttt tataatgcca
     2101 actttgtaca agaaagctgg gtcgaattcg cccttcggtc tcaggagctt catcaagaga
     2161 gcaaccaaca aagaggaggc gaacacggtg cacagcaccg tctgcatact tccgatttgc
     2221 aattcgcaac atatgataca tcatccatct ctctgagaaa actgggcact gttagaatct
     2281 gggatttatc aacagccacc aactatctac tcaacgcaat tggggcactt gataatgtga
     2341 caatcctcct tgctcctttc cttgtcgcct caaacaacgt cgggaactgc acagaaacat
     2401 cgctacttat tcacgcctca cctctctttc ctcctttact cacttattcg cgcctcacct
     2461 ctctttcctc ctttactcaa gtcctctgct tgccgggttt attcttccgt ttgtgttgtt
     2521 gcggtctgat aggatttgtt tggagacgaa cctttttttt tgattgcttt ataaattaac
     2581 gttttgccga aattgttgag acctaagggc gaattcggag cctgcttttt tgtacaaagt
     2641 tggcattata aaaaagcatt gctcatcaat ttgttgcaac gaacaggtca ctatcagtca
     2701 aaataaaatc attatttggg gcccgagctt aagactggcc gtcgttttac aacgtcgtga
     2761 ctgggaaaac atccatgcta gcgttaacgc gagagtaggg aactgccagg catcaaataa
     2821 aacgaaaggc tcagtcggaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2881 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgtgaag caacggcccg
     2941 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaactaag cagaaggcca
     3001 tcctgacgga tggccttttt gcgtttctac aaactcttcc tggctagcgg tacgcgtatt
     3061 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3121 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3181 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3241 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaag
//
