LOCUS       sequence-211192-        4864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pKO but without the negative selection gene galK..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4864)
  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 4864)
  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: pKOdeltagalK (Hyg)
FEATURES             Location/Qualifiers
     source          1..4864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1785..1807
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1799..1816
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1800..1816
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2060..2648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2140..2159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2793..3238
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(3239..3325,3326..4660)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
       61 ttcatccata gttgcctgac tccccgacgt ggccgaccag cccgtcatcg tcaacgcctg
      121 atccgcggtg cggacaggcc gtgtcgcgac cggccgtgcg gaattaagcc ggcccgtacc
      181 ctgtgaatag aggtccgctg tgacacaaga atccctgtta cttctcgacc gtattgattc
      241 ggatgattcc tacgcgagcc tgcggaacga ccaggaattc tgggagccgc tggcccgccg
      301 agccctggag gagctcgggc tgccggtgcc gccggtgctg cgggtgcccg gcgagagcac
      361 caaccccgta ctggtcggcg agcccggccc ggtgatcaag ctgttcggcg agcactggtg
      421 cggtccggag agcctcgcgt cggagtcgga ggcgtacgcg gtcctggcgg acgccccggt
      481 gccggtgccc cgcctcctcg gccgcggcga gctgcggccc ggcaccggag cctggccgtg
      541 gccctacctg gtgatgagcc ggatgaccgg caccacctgg cggtccgcga tggacggcac
      601 gaccgaccgg aacgcgctgc tcgccctggc ccgcgaactc ggccgggtgc tcggccggct
      661 gcacagggtg ccgctgaccg ggaacaccgt gctcaccccc cattccgagg tcttcccgga
      721 actgctgcgg gaacgccgcg cggcgaccgt cgaggaccac cgcgggtggg gctacctctc
      781 gccccggctg ctggaccgcc tggaggactg gctgccggac gtggacacgc tgctggccgg
      841 ccgcgaaccc cggttcgtcc acggcgacct gcacgggacc aacatcttcg tggacctggc
      901 cgcgaccgag gtcaccggga tcgtcgactt caccgacgtc tatgcgggag actcccgcta
      961 cagcctggtg caactgcatc tcaacgcctt ccggggcgac cgcgagatcc tggccgcgct
     1021 gctcgacggg gcgcagtgga agcggaccga ggacttcgcc cgcgaactgc tcgccttcac
     1081 cttcctgcac gacttcgagg tgttcgagga gaccccgctg gatctctccg gcttcaccga
     1141 tccggaggaa ctggcgcagt tcctctgggg gccgccggac accgcccccg gcgcctgacg
     1201 ccccgggccc gcgccgaagc ccgctgctgc gagcccggag cgggccggcc gacggcggtg
     1261 cgggcccggc ggcggacgct cagcagcggc gggcgtgaaa ggccctggca tcctcgatca
     1321 tctcctccag ggtgggtcgg ccgggcttcc atcccagctc ggcaaggatc aacgaggaca
     1381 gtcgcacgac gaagttcttc tggatcgcgc ccgtgctgga agcactcaac ctcgaagcgt
     1441 gtggttgcgg agccatctag caaccacacg accatggatc ctcccgctgc gctgagtctt
     1501 cagcgatccc catccgcgac gtgccaacta ggtctcctct cgtcgtgaac aaggctaccg
     1561 ggttgcaact cctgtgcaac tctcaggctt caacgcgctt ctacgacctg caatttcttt
     1621 ccacttagag gatgcagccg agagggggta aaaacctccc attcgccatt caggctgcgc
     1681 aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     1741 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     1801 taaaacgacg gccagtgagc gcgacgtaat aagttgaaga gcatctgctc ttcagcatca
     1861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg tcatagctgt
     1921 ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1981 cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2461 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2581 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2641 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaggcg
     2701 cgcctgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2761 gcgaggaagc ggaagatcgg gcccagccgg cccacatata cctgccgttc actattattt
     2821 agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac tttatgccca
     2881 tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt ttttagttct
     2941 ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga ggaaaataga
     3001 ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt
     3061 tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg
     3121 tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa
     3181 caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgat
     3241 gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg cactgctggc
     3301 aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata aggaaacata
     3361 cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc aaaaaaatga
     3421 aaaatataaa gttcctgagt tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg
     3481 cctggacgtt tgggacagct ggccattaca aaacactgac ggcactgtcg caaactatca
     3541 cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg acacatcgat
     3601 ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa acgctggccg
     3661 cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag accaaacaca
     3721 agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat tctacactga
     3781 tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta acgtatcagc
     3841 atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct ttgacggtga
     3901 cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca gctcaggcga
     3961 caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat acttagtatt
     4021 tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat ttaacaaagc
     4081 atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc tgcaaagcga
     4141 taaaaaacgc acggctgagt tagcaaacgg cgctctcggt atgattgagc taaacgatga
     4201 ttacacactg aaaaaagtga tgaaaccgct gattgcatct aacacagtaa cagatgaaat
     4261 tgaacgcgcg aacgtcttta aaatgaacgg caaatggtac ctgttcactg actcccgcgg
     4321 atcaaaaatg acgattgacg gcattacgtc taacgatatt tacatgcttg gttatgtttc
     4381 taattcttta actggcccat acaagccgct gaacaaaact ggccttgtgt taaaaatgga
     4441 tcttgatcct aacgatgtaa cctttactta ctcacacttc gctgtacctc aagcgaaagg
     4501 aaacaatgtc gtgattacaa gctatatgac aaacagagga ttctacgcag acaaacaatc
     4561 aacgtttgcg cctagcttcc tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga
     4621 cagcatcctt gaacaaggac aattaacagt taacaaataa aaacgcaaaa gaaaatgccg
     4681 atatcctatt ggcattgcgg ccgcggtctc ggcggtgggt cgtctagagg tctgacgctc
     4741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4861 cttg
//
