LOCUS       sequence-140781-         917 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  ""intermediate vector"" for the two-step cloning method (Wang and 
            Szaro, Dev Biol, 2015).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 917)
  AUTHORS   Wang C, Szaro BG
  TITLE     A method for using direct injection of plasmid DNA to study 
            cis-regulatory element activity in F0 Xenopus embryos and tadpoles.
  JOURNAL   Dev Biol. 2015 Feb 1;398(1):11-23. doi: 10.1016/j.ydbio.2014.11.010.
            Epub 2014 Nov 20.
  PUBMED    25448690
REFERENCE   2  (bases 1 to 917)
  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"; doi: 
            "10.1016/j.ydbio.2014.11.010"; journalName: "Dev Biol"; date: 
            "2015-02-1- 1"; volume: "398"; issue: "1"; pages: "11-23"
COMMENT     Sequence Label: pBluescriptII[attB/Ins1]
FEATURES             Location/Qualifiers
     source          1..917
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     215..232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    349..370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(385..390,391..408,409..415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    423..439
                     /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     428..450
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     447..463
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     447..463
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     482..502
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        484..502
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
       61 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      181 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      241 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      301 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      361 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      421 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
      481 cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgctc
      541 tagaactagt gcgacagagc gataaagctt tttccccgta tccccccagg tgtctgcagg
      601 ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccg
      661 ggctgtcccc gcacgctgcg ggctccggct cggggatgcg gggggagcgt cgggccggag
      721 cggagccccg ggcggctcgc tgctgccccc tagcgggaga ggaacgcaat tacagccctg
      781 ggggctttgg gggggggctg tccccgtgag ctcccgcgga tctagataaa gctttttccc
      841 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
      901 tcccgtgcca ccttccc
//
