LOCUS       sequence-236334-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 Part. CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Vasudevan R, Gale GAR, Schiavon AA, Puzorjov A, Malin J, Gillespie 
            MD, Vavitsas K, Zulkower V, Wang B, Howe CJ, Lea-Smith DJ, McCormick
            AJ
  TITLE     CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria 
            Based on the Plant MoClo Syntax.
  JOURNAL   Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub
            2019 Feb 28.
  PUBMED    30819783
REFERENCE   2  (bases 1 to 6357)
  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"; doi: 
            "10.1104/pp.18.01401"; journalName: "Plant Physiol"; date: 
            "2019-05"; volume: "180"; issue: "1"; pages: "39-55"
COMMENT     Sequence Label: pC0.017
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        110..214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             436..1227
                     /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"
     rep_origin      1398..1986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1887..1906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2140..2157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(2235..2261,2262..2264,2265..4751,4752..4754,
                     4755..6341)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
       61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
      241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      301 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
      361 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
      421 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
      481 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
      541 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
      601 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
      661 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
      721 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
      781 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
      841 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
      901 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
      961 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     1021 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     1081 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     1141 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
     1201 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
     1261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
     2221 ctctgtggtc tcaaatggat aagaaatact caataggctt agctatcggc acaaatagcg
     2281 tcggatgggc ggtgatcact gatgaatata aggttccgtc taaaaagttc aaggttctgg
     2341 gaaatacaga ccgccacagt atcaaaaaaa atcttatagg ggctctttta tttgacagtg
     2401 gagagacagc ggaagcgact cgtctcaaac ggacagctcg tagaaggtat acacgtcgga
     2461 agaatcgtat ttgttatcta caggagattt tttcaaatga gatggcgaaa gtagatgata
     2521 gtttctttca tcgacttgaa gagtcttttt tggtggaaga agataagaag catgaacgtc
     2581 atcctatttt tggaaatata gtagatgaag ttgcttatca tgagaaatat ccaactatct
     2641 atcatctgcg aaaaaaattg gtagattcta ctgataaagc ggatttgcgc ttaatctatt
     2701 tggccttagc gcatatgatt aagtttcgtg gtcatttttt gattgaggga gatttaaatc
     2761 ctgataatag tgatgtggac aaactattta tccagttggt acaaacctac aatcaattat
     2821 ttgaagaaaa ccctattaac gcaagtggag tagatgctaa agcgattctt tctgcacgat
     2881 tgagtaaatc aagacgatta gaaaatctca ttgctcagct ccccggtgag aagaaaaatg
     2941 gcttatttgg gaatctcatt gctttgtcat tgggtttgac ccctaatttt aaatcaaatt
     3001 ttgatttggc agaagatgct aaattacagc tttcaaaaga tacttacgat gatgatttag
     3061 ataatttatt ggcgcaaatt ggagatcaat atgctgattt gtttttggca gctaagaatt
     3121 tatcagatgc tattttactt tcagatatcc taagagtaaa tactgaaata actaaggctc
     3181 ccctatcagc ttcaatgatt aaacgctacg atgaacatca tcaagacttg actcttttaa
     3241 aagctttagt tcgacaacaa cttccagaaa agtataaaga aatctttttt gatcaatcaa
     3301 aaaacggata tgcaggttat attgatgggg gagctagcca agaagaattt tataaattta
     3361 tcaaaccaat tttagaaaaa atggatggta ctgaggaatt attggtgaaa ctaaatcgtg
     3421 aagatttgct gcgcaagcaa cggacctttg acaacggctc tattccccat caaattcact
     3481 tgggtgagct gcatgctatt ttgagacgac aagaagattt ttatccattt ttaaaagaca
     3541 atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc ttattatgtt ggtccattgg
     3601 cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca attaccccat
     3661 ggaattttga agaagttgtc gataaaggtg cttcagctca atcatttatt gaacgcatga
     3721 caaactttga taaaaatctt ccaaatgaaa aagtactacc aaaacatagt ttgctttatg
     3781 agtattttac ggtttataac gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa
     3841 aaccagcatt tctttcaggt gaacagaaga aagccattgt tgatttactc ttcaaaacaa
     3901 atcgaaaagt aaccgttaag caattaaaag aagattattt caaaaaaata gaatgttttg
     3961 atagtgttga aatttcagga gttgaagata gatttaatgc ttcattaggt acctaccatg
     4021 atttgctaaa aattattaaa gataaagatt ttttggataa tgaagaaaat gaagatatct
     4081 tagaggatat tgttttaaca ttgaccttat ttgaagatag ggagatgatt gaggaaagac
     4141 ttaaaacata tgctcacctc tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt
     4201 atactggttg gggacgtttg tctcgaaaat tgattaatgg tattagggat aagcaatctg
     4261 gcaaaacaat attagatttt ttgaaatcag atggttttgc caatcgcaat tttatgcagc
     4321 tgatccatga tgatagtttg acatttaaag aagatattca aaaagcacaa gtgtctggac
     4381 aaggcgatag tttacatgaa catattgcaa atttagctgg tagccctgct attaaaaaag
     4441 gtattttaca gactgtaaaa gttgttgatg aattggtcaa agtaatgggg cggcataagc
     4501 cagaaaatat cgttattgaa atggcacgtg aaaatcagac aactcaaaag ggccagaaaa
     4561 attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga agtcagattc
     4621 ttaaagagca tcctgttgaa aatactcaat tgcaaaatga aaagctctat ctctattatc
     4681 tccaaaatgg aagagacatg tatgtggacc aagaattaga tattaatcgt ttaagtgatt
     4741 atgatgtcga tgccattgtt ccacaaagtt tccttaaaga cgattcaata gacaataagg
     4801 tcttaacgcg ttctgataaa aatcgtggta aatcggataa cgttccaagt gaagaagtag
     4861 tcaaaaagat gaaaaactat tggagacaac ttctaaacgc caagttaatc actcaacgta
     4921 agtttgataa tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat aaagctggtt
     4981 ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa gcatgtggca caaattttgg
     5041 atagtcgcat gaatactaaa tacgatgaaa atgataaact tattcgagag gttaaagtga
     5101 ttaccttaaa atctaaatta gtttctgact tccgaaaaga tttccaattc tataaagtac
     5161 gtgagattaa caattaccat catgcccatg atgcgtatct aaatgccgtc gttggaactg
     5221 ctttgattaa gaaatatcca aaacttgaat cggagtttgt ctatggtgat tataaagttt
     5281 atgatgttcg taaaatgatt gctaagtctg agcaagaaat aggcaaagca accgcaaaat
     5341 atttctttta ctctaatatc atgaacttct tcaaaacaga aattacactt gcaaatggag
     5401 agattcgcaa acgccctcta atcgaaacta atggggaaac tggagaaatt gtctgggata
     5461 aagggcgaga ttttgccaca gtgcgcaaag tattgtccat gccccaagtc aatattgtca
     5521 agaaaacaga agtacagaca ggcggattct ccaaggagtc aattttacca aaaagaaatt
     5581 cggacaagct tattgctcgt aaaaaagact gggatccaaa aaaatatggt ggttttgata
     5641 gtccaacggt agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga
     5701 agttaaaatc cgttaaagag ttactaggga tcacaattat ggaaagaagt tcctttgaaa
     5761 aaaatccgat tgacttttta gaagctaaag gatataagga agttaaaaaa gacttaatca
     5821 ttaaactacc taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta
     5881 gtgccggaga attacaaaaa ggaaatgagc tggctctgcc aagcaaatat gtgaattttt
     5941 tatatttagc tagtcattat gaaaagttga agggtagtcc agaagataac gaacaaaaac
     6001 aattgtttgt ggagcagcat aagcattatt tagatgagat tattgagcaa atcagtgaat
     6061 tttctaagcg tgttatttta gcagatgcca atttagataa agttcttagt gcatataaca
     6121 aacatagaga caaaccaata cgtgaacaag cagaaaatat tattcattta tttacgttga
     6181 cgaatcttgg agctcccgct gcttttaaat attttgatac aacaattgat cgtaaacgat
     6241 atacgtctac aaaagaagtt ttagatgcca ctcttatcca tcaatccatc actggtcttt
     6301 atgaaacacg cattgatttg agtcagctag gaggtgacta agctttgaga ccacgaa
//
