LOCUS       sequence-139621-        2229 bp    DNA     circular SYN 14-APR-2025
DEFINITION  E. coli cloning vector (KanR, low copy, blue/white selection).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2229)
  AUTHORS   Vieira J, Messing J
  TITLE     New pUC-derived cloning vectors with different selectable markers 
            and DNA replication origins.
  JOURNAL   Gene. 1991 Apr;100:189-94.
  PUBMED    1905257
REFERENCE   2  (bases 1 to 2229)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene."; 
            date: "1991-04"; volume: "100"; pages: "189-94"
COMMENT     Sequence Label: pOK12
FEATURES             Location/Qualifiers
     source          1..2229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(51..595)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(131..150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(794..813)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             805..1614
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(872..891)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1681..2025)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPLVRGLEIYRCMPWYPGARIRSFCRRVDVIWIRYRRGGRS
                     RTSGSIPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRP
                     SQQLRSLNGE"
     primer_bind     1806..1828
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1820..1837
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1821..1837
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1844..1863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1844..1862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1877..1893)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1877..1893)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2021..2037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2021..2037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2034..2056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2045..2061
                     /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)."
     protein_bind    2116..2137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattgga gctttcgcga gggaagtgag agggccgcgg caaagccgtt tttccatagg
       61 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
      181 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
      241 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
      301 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
      361 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtttgaa
      421 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
      481 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
      541 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
      601 tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcctagagtc gagggggggg ggggaaagcc acgttgtgtc tcaaaatctc
      721 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
      781 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
      841 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
      901 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
      961 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1021 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1081 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     1141 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1201 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     1261 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1321 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     1381 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1441 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     1501 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     1561 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     1621 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     1681 tcattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     1741 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     1801 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
     1861 gggcgaattg gggatcgatc cactagttct agagcggccg ccacggcgat atcggatcca
     1921 tatgacgtcg acgcgtctgc agaagcttcg aattcgagct cccgggtacc atggcatgca
     1981 tcgatagatc tcgaggcctc ggactagtgg cgtaatcatg gtcatagctg tttcctgtgt
     2041 gaaattgtta tccgctcaca attccacaca acatacgagc cgcggaagca taaagtgtaa
     2101 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2161 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2221 aggcggttt
//
