LOCUS       sequence-74217-e         972 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 972)
  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 972)
  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: pICH41246
FEATURES             Location/Qualifiers
     source          1..972
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    119..140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(155..160,161..178,179..185)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    193..209
                     /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     198..220
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     217..233
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     217..233
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             229..552
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    246..302
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(303..320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(303..319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(312..334)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(528..547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(689..707)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        775..879
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaccgcctct ccccgcgcgt tggccgattc attaatcact ctgtggtctc atactttgtc
       61 ttcgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      121 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      181 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
      241 acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat
      301 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
      361 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
      421 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
      481 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
      541 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      601 gcttgtctgc tcccggcatc cgcttacaga caagctgtga cgaagacaaa atgtgagacc
      661 acgaagtggc tcttcagtgg acgaaagggc ctcgtgatac gcctattttt ataggttaat
      721 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgcgctcacg caactggtcc
      901 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
      961 gactgttttt tt
//
