LOCUS       sequence-90432-e        1003 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  cloning backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1003)
  AUTHORS   Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu 
            J, Jiang W, Yang S
  TITLE     High-Efficiency Scarless Genetic Modification in Escherichia coli by
            Using Lambda Red Recombination and I-SceI Cleavage.
  JOURNAL   Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr 
            18.
  PUBMED    24747889
REFERENCE   2  (bases 1 to 1003)
  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: "Appl 
            Environ Microbiol."; date: "2014-07-1"; volume: "80(13)"; pages: 
            "3826-3834. Epub 2014 Apr 18"
COMMENT     Sequence Label: pKSI-1
FEATURES             Location/Qualifiers
     source          1..1003
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     28..47
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        28..46
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    55..162
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     79..95
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     79..95
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(129..145)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(130..146)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(175..193)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(250..266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(250..266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(263..285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    274..290
                     /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(298..304,305..322,323..328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    343..364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(481..498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(732..751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatccctag ccagcgcttt gatggtgtaa tacgactcac tatagggcga attggagctc
       61 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga attcgatatc
      121 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccagctttt gttcccttta
      181 gtgagggtta attggccagc agaggcctgg tagggataac agggtaattg cgcgcttggc
      241 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      301 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      361 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      421 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      841 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      961 tgagtccaac ccggtaagac acgacttatc gccactggca gca
//
