LOCUS       sequence-74277-e         993 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 993)
  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 993)
  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: pICH75377
FEATURES             Location/Qualifiers
     source          1..993
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    128..149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(164..169,170..187,188..194)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    202..218
                     /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     207..229
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     226..242
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     226..242
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             238..561
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    255..311
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(312..329)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(312..328)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(321..343)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(537..556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    688..712
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     complement(915..934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cattgcggac gtttttaatg tactggggtg gatgcagtgg gccccactct gtggaagaca
       61 atgtgtgaga ccgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      121 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
      181 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
      241 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
      301 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      361 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      421 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
      481 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
      541 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
      601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgag gtctcaggga
      661 cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga
      721 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg
      781 tccatttgta tgtgccaacc acaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      841 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      901 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      961 tgttccgacc ctgccgctta ccggatacct gtc
//
