LOCUS       sequence-75159-e         822 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 822)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S
  TITLE     A modular cloning system for standardized assembly of multigene 
            constructs.
  JOURNAL   PLoS One. 2011 Feb 18;6(2):e16765. doi: 
            10.1371/journal.pone.0016765.
  PUBMED    21364738
REFERENCE   2  (bases 1 to 822)
  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: "PLoS One.";
            date: "2011-02-18"; volume: "6(2):e16765. doi"; pages: " 
            10.1371/journal.pone.0016765"
COMMENT     Sequence Label: pICH49255
FEATURES             Location/Qualifiers
     source          1..822
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    27..51
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(186..509)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     191..210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     404..426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     418..435
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     419..435
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    436..492
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(505..521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(505..521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(518..540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    529..545
                     /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)."
     promoter        complement(join(553..559,560..577,578..583))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    598..619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    774..798
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc aaacactgat
       61 agtttaaacc acttcgtgaa gacaatccct gagaccgtca cagcttgtct gtaagcggat
      121 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
      181 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
      241 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
      301 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
      361 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
      421 aaaacgacgg ccagtgaatt cgagctcggt acccggggat cctctagagt cgacctgcag
      481 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      541 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      661 gtcgtgccag ctgcggtctc attgcttgtc ttcacagagt ggggcccact gcatccaccc
      721 cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca
      781 ccacaatata tcctgccacc agccagccaa cagctccccg ac
//
