LOCUS       sequence-74325-e         996 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 996)
  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 996)
  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: pICH47841
FEATURES             Location/Qualifiers
     source          1..996
                     /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(193..516)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     198..217
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     411..433
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     425..442
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     426..442
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    443..499
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(512..528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(512..528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(525..547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    536..552
                     /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(560..566,567..584,585..590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    605..626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    797..821
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc aaacactgat
       61 agtttaaacc acttcgtgca gaagacaaca caggagtgag accgtcacag cttgtctgta
      121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
      181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
      241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
      301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      361 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      421 acgttgtaaa acgacggcca gtgaattcga gctcggtacc cggggatcct ctagagtcga
      481 cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      661 gaaacctgtc gtgccagctg cggtctcacg ctgcatgcga attctctgtt gtcttcacag
      721 agtggggccc actgcatcca ccccagtaca ttaaaaacgt ccgcaatgtg ttattaagtt
      781 gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc
      841 ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca gtcagatcag
      901 gatctccttt gcgacgctca ccgggctggt tgccctcgcc gctgggctgg cggccgtcta
      961 tggccctgca aacgcgccag aaacgccgtc gaagcc
//
