LOCUS       sequence-406995-        2987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 acceptor vector for assembling gRNA arrays for Cas12a in SP 
            position .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2987)
  AUTHORS   Victoria AJ, Selao TT, Moreno-Cabezuelo JA, Mills LA, Gale GAR, 
            Lea-Smith DJ, McCormick AJ
  TITLE     A toolbox to engineer the highly productive cyanobacterium 
            Synechococcus sp. PCC 11901.
  JOURNAL   Plant Physiol. 2024 May 7:kiae261. doi: 10.1093/plphys/kiae261.
  PUBMED    38713768
REFERENCE   2  (bases 1 to 2987)
  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 
            Physiol. 2024 May 7:kiae261. doi: 10.1093/plphys/kiae261."
COMMENT     Sequence Label: pCA0.421
FEATURES             Location/Qualifiers
     source          1..2987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             398..1189
                     /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      1360..1948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1849..1868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2102..2119
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2200..2234
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     protein_bind    2319..2340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2355..2360,2361..2378,2379..2385)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2393..2409
                     /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     2398..2420
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2417..2433
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2417..2433
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2429..2752
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPGLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     complement(2503..2520)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2503..2519)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2512..2534)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2728..2747)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(join(2973..2987,1..4))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
       61 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatgc
      181 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
      241 ttttcatggc ttgttatgac tgtttttttt tggggtacag tctatgcctc gggcatccaa
      301 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
      361 gcagcagggc agtcgcccta aaacaaagtt aaacatcatg agggaagcgg tgatcgccga
      421 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
      481 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagcgatat
      541 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
      601 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
      661 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
      721 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
      781 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
      841 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
      901 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
      961 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     1021 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     1081 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     1141 atttgtccat tacgtaaaag gcgagatcac caaggtagtc ggcaaataac tgtcagacca
     1201 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1261 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1501 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1741 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2041 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2101 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2161 gcgttggccg attcattaat cactctgtgg tctcaaatgt tgacagctag ctcagtccta
     2221 ggtataatgc tagcagctga tttaggcaaa aacggttgtc ttcgcagctg gcacgacagg
     2281 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     2341 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     2401 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaggct tgcatgcctg
     2461 caggtcgact ctagaggatc cccgggtacc gagctcgaat tcactggccg tcgttttaca
     2521 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2581 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2641 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2701 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2761 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2821 cgcttacaga caagctgtga cgaagacaaa acgaatttct actgttgtag atctcggtac
     2881 caaattccag aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg tccaggttga
     2941 gaccacgaag tggctcttca gtggacgaaa gggcctcgtg atacgcc
//
