LOCUS       sequence-211172-        3947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pKO but without the negative selection gene sacB..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3947)
  AUTHORS   Arnold FM, Hohl M, Remm S, Koliwer-Brandl H, Adenau S, Chusri S, 
            Sander P, Hilbi H, Seeger MA
  TITLE     A uniform cloning platform for mycobacterial genetics and protein 
            production.
  JOURNAL   Sci Rep. 2018 Jun 22;8(1):9539. doi: 10.1038/s41598-018-27687-5.
  PUBMED    29934571
REFERENCE   2  (bases 1 to 3947)
  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: "Sci Rep."; 
            date: "2018-06-22"; volume: "8(1):9539. doi"; pages: " 
            10.1038/s41598-018-27687-5"
COMMENT     Sequence Label: pKOdeltasacB (Kan)
FEATURES             Location/Qualifiers
     source          1..3947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(92..97,98..115,116..122)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    128..147
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     1738..1754
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1888..1905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1998..2586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2078..2097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2691..2707
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2717..2736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2717..2735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3056..3871)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3779..3798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3863..3882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 taacatcaga gattttgaga cacaacgtgg ctttcccccc ccccgacgtc aggtggctag
       61 aactagatat ccatggatcc agaccccagg cttgacactt tatgcttccg gctcgtataa
      121 tgtgtggaat tgtgagcgct cacaattcgc atgcggtacc aagctgatcc gataacacag
      181 gaacagatct tccggagtgt aagaaatgag tctgaaagaa aaaacacaat ctctgtttgc
      241 caacgcattt ggctaccctg ccactcacac cattcaggcg cctggccgcg tgaatttgat
      301 tggtgaacac accgactaca acgacggttt cgttctgccc tgcgcgattg attatcaaac
      361 cgtgatcagt tgtgcaccac gcgatgaccg taaagttcgc gtgatggcag ccgattatga
      421 aaatcagctc gacgagtttt ccctcgatgc gcccattgtc gcacatgaaa actatcaatg
      481 ggctaactac gttcgtggcg tggtgaaaca tctgcaactg cgtaacaaca gcttcggcgg
      541 cgtggacatg gtgatcagcg gcaatgtgcc gcagggtgcc gggttaagtt cttccgcttc
      601 actggaagtc gcggtcggaa ccgtattgca gcagctttat catctgccgc tggacggcgc
      661 acaaatcgcg cttaacggtc aggaagcaga aaaccagttt gtaggctgta actgcgggat
      721 catggatcag ctaatttccg cgctcggcaa gaaagatcat gccttgctga tcgattgccg
      781 ctcactgggg accaaagcag tttccatgcc caaaggtgtg gctgtcgtca tcatcaacag
      841 taacttcaaa cgtaccctgg ttggcagcga atacaacacc cgtcgtgaac agtgcgaaac
      901 cggtgcgcgt ttcttccagc agccagccct gcgtgatgtc accattgaag agttcaacgc
      961 tgttgcgcat gaactggacc cgatcgtggc aaaacgcgtg cgtcatatac tgactgaaaa
     1021 cgcccgcacc gttgaagctg ccagcgcgct ggagcaaggc gacctgaaac gtatgggcga
     1081 gttgatggcg gagtctcatg cctctatgcg cgatgatttc gaaatcaccg tgccgcaaat
     1141 tgacactctg gtagaaatcg tcaaagctgt gattggcgac aaaggtggcg tacgcatgac
     1201 cggcggcgga tttggcggct gtatcgtcgc gctgatcccg gaagaactgg tgcctgccgt
     1261 acagcaagct gtcgctgaac aatatgaagc aaaaacaggt attaaagaga ctttttacgt
     1321 ttgtaaacca tcacaaggag caggacagtg ctgaacgaaa ctcctgcagc gcgtagtacc
     1381 tcacaccgtc ctgcgggttg acgtatgagg cttggatcct cccgctgcgc tgagtcttca
     1441 gcgatcccca tccgcgacgt gccaactagg tctcctctcg tcgtgaacaa ggctaccggg
     1501 ttgcaactcc tgtgcaactc tcaggcttca acgcgcttct acgacctgca atttctttcc
     1561 acttagagga tgcagccgag agggggtaaa aacctcccat tcgccattca ggctgcgcaa
     1621 ctgttgggaa gggcgtttcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     1681 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     1741 aaacgacggc cagtgagcgc gacgtaataa gttgaagagc atctgctctt cagcatcact
     1801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taacatggtc atagctgttt
     1861 ccttgcgtat tgggcgctct ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1921 ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1981 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2041 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2101 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2161 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2221 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2281 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2341 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2401 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2461 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2521 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2581 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2641 cgttaaggga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     2701 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctcc accgcggtgg
     2761 cggccgctct agtgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga
     2821 atcgccccat catccagcca gaaagtgagg gagccacggt tggtgagagc tttgttgtag
     2881 gtggaccagt tggtgatttt gaacttttgc tttgccacgg aatggtctgc gttgtcggga
     2941 agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt
     3001 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     3061 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     3121 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     3181 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     3241 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     3301 ccggtgagaa tggcaaaaga ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3361 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     3421 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     3481 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     3541 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3601 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3661 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3721 ctggcgcatg gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat
     3781 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     3841 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     3901 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatg
//
