LOCUS       sequence-234014-        2855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 acceptor vector for promoters truncated to transcriptional 
            start site with lacZ compatible with blue/white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2855)
  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 2855)
  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: pCA0.001
FEATURES             Location/Qualifiers
     source          1..2855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..58)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        126..230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             452..1243
                     /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      1414..2002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1903..1922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2156..2173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2321..2342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2357..2362,2363..2380,2381..2387)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2395..2411
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2400..2422
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2419..2435
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2419..2435
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2431..2754
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2448..2504
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2505..2522)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2505..2521)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2514..2536)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2730..2749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tagcagagac cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt
       61 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgcgctcac
      241 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
      301 tggcttgtta tgactgtttt tttttggggt acagtctatg cctcgggcat ccaagcagca
      361 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
      421 gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc
      481 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
      541 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagcg atattgattt
      601 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
      661 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
      721 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
      781 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
      841 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
      901 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
      961 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     1021 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     1081 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     1141 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     1201 ccattacgta aaaggcgaga tcaccaaggt agtcggcaaa taactgtcag accaagttta
     1261 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1321 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1381 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1441 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1501 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1561 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1621 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1681 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1741 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1801 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1861 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1921 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcttc
     1981 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2041 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2101 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2161 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2221 gccgattcat taatcactct gtggtctctg gaggagtctt cgtacgcagc tggcacgaca
     2281 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2341 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2401 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc
     2461 tgcaggtcga ctctagagga tccccgggta ccgagctcga attcactggc cgtcgtttta
     2521 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2581 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2641 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2701 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2761 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2821 tccgcttaca gacaagctgt gacgtacgaa gactc
//
