LOCUS       sequence-236339-        4835 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 4835)
  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 4835)
  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: pC0.026
FEATURES             Location/Qualifiers
     source          1..4835
                     /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"
     promoter        2238..2683
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(2684..2770,2771..4105)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
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 tcaaatgcac atatacctgc cgttcactat tatttagtga aatgagatat
     2281 tatgatattt tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat
     2341 aaaaaataca gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct
     2401 gcaaatcctt ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa
     2461 acgagagtct aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc
     2521 aggcaagacc taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat
     2581 tttaggtctt tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag
     2641 aagcagaccg ctaacacagt acataaaaaa ggagacatga acgatgaaca tcaaaaagtt
     2701 tgcaaaacaa gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca
     2761 agcgtttgcg aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat
     2821 tacacgccat gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagttcc
     2881 tgaattcgat tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg acgtttggga
     2941 cagctggcca ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt
     3001 ctttgcatta gccggagatc ctaaaaatgc ggatgacaca tcgatttaca tgttctatca
     3061 aaaagtcggc gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag
     3121 cgacaaattc gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc
     3181 agccacattt acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca
     3241 ttacggcaaa caaacactga caactgcaca agttaacgta tcagcatcag acagctcttt
     3301 gaacatcaac ggtgtagagg attataaatc aatctttgac ggtgacggaa aaacgtatca
     3361 aaatgtacag cagttcatcg atgaaggcaa ctacagctca ggcgacaacc atacgctgag
     3421 agatcctcac tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg
     3481 aactgaagat ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag
     3541 cacatcattc ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc
     3601 tgagttagca aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa
     3661 agtgatgaaa ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt
     3721 ctttaaaatg aacggcaaat ggtacctgtt cactgactcc cgcggatcaa aaatgacgat
     3781 tgacggcatt acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg
     3841 cccatacaag ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga
     3901 tgtaaccttt acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat
     3961 tacaagctat atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgccaag
     4021 cttcctgctg aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca
     4081 aggacaatta acagttaaca aataaaaacg caaaagaaaa tgccgatatc ctattggcat
     4141 tttcttttat ttcttatcaa cataaaggtg aatcccatat gaactatata aaagcaggca
     4201 aatggctaac cgtattccta accttttggt aatgactcca acttattgat agtgttttat
     4261 gttcagataa tgcccgatga ctttgtcatg cagctccacc gattttgaga acgacagcga
     4321 cttccgtccc agccgtgcca ggtgctgcct cagattcagg ttatgccgct caattcgctg
     4381 cgtatatcgc ttgctgatta cgtgcagctt tcccttcagg cgggattcat acagcggcca
     4441 gccatccgtc atccatatca ccacgtcaaa gggtgacagc aggctcataa gacgccccag
     4501 cgtcgccata gtgcgttcac cgaatacgtg cgcaacaacc gtctaccgga gactgtcata
     4561 cgcgtaaaac agccagcgct ggcgcgattt agccccgaca tagccccact gttcgtccat
     4621 ttccgcgcag acgatgacgt cactgcccgg ctgtatgcgc gaggttaccg actgcggcct
     4681 gagtttttta agtgacgtaa aatcgtgttg aggccaacgc ccataatgcg ggctgttgcc
     4741 cggcatccaa cgccattcat ggccatatca atgattttct ggtgcgtacc gggttgagaa
     4801 gcggtgtaag tgaactgcaa ggttgagacc acgaa
//
