LOCUS       sequence-418045-        4817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L5 integrating vector for delivery of defective Hyg gene (ZeoR) for 
            oligonucleotide-mediated SNP transfer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4817)
  AUTHORS   Murphy KC
  TITLE     Oligo-Mediated Recombineering and its Use for Making SNPs, 
            Knockouts, Insertions, and Fusions in Mycobacterium tuberculosis.
  JOURNAL   Methods Mol Biol. 2021;2314:301-321. doi: 
            10.1007/978-1-0716-1460-0_14.
  PUBMED    34235660
REFERENCE   2  (bases 1 to 4817)
  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: "Methods Mol
            Biol."; date: "2021"; volume: "2314:301-321. doi"; pages: " 
            10.1007/978-1-0716-1460-0_14"
COMMENT     Sequence Label: pKM427
FEATURES             Location/Qualifiers
     source          1..4817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(767..1355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(847..866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1887..2261)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(2280..2327)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     terminator      complement(4153..4199)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(4218..4320)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gggtgagcac ggtgttcccg gtcagcggca ccctgtgcag ccggccgagc acccggccga
       61 gttcgcgggc cagggcgagc agcgcgttcc ggtcggtcgt gccgtctatt gcggaccgcc
      121 aggtggtgcc ggtcatccgg ctcatcacca ggtagggcca cggccaggct ccggtgccgg
      181 gccgcagctc gccgcggccg aggaggcggg gcaccggcac cggggcgtcc gccaggaccg
      241 cgtacgcctc cgactccgac gcgaggctct ccggaccgca ccagtgctcg ccgaacagct
      301 tgatcaccgg gccgggctcg ccgaccagta cggggttggt gctctcgccg ggcacccgca
      361 gcaccggcgg caccggtagc ccgagctcct ccagggctcg gcgggccagc ggctcccaga
      421 attcctggtc gttccgcagg ctcgcgtagg aatcatccga atcaatacgg tcgagaagta
      481 acagggattc ttgtgtcaca gcggacctct attcacaggg tacgggccgg cttaattccg
      541 cacggccggt cgcgacacgg cctgtccgca ccgcggatca ggcgttgacg atgacgggct
      601 ggtcggccac gtcggggagt aagcttaagc aacggcccca gtgagtagcg actttggatc
      661 cggtacccgc aacggtcaac cagcgtctac gcgtttcact ggccgacatg tgagcaaaag
      721 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      781 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      841 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      901 ccctgccgct taccggatac ctgtccgcct ttctcccctc gggaagcgtg gcgctttctc
      961 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1021 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1081 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1141 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1201 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1261 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1321 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1381 ggtctgacgc tcatggaact agtgaggtct gcctcgtgaa gaaggtgttg ctgactcata
     1441 ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt tggtgagagc
     1501 tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc
     1561 gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca
     1621 aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat
     1681 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1741 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1801 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1861 caacctatta tggagctcgg gaattctcag tcctgctcct cggccacgaa gtgcacgcag
     1921 ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc
     1981 atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac
     2041 agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc
     2101 tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc
     2161 acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg
     2221 cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtttagtt cctcaccttg
     2281 tcgtattata ctatgccgat atactatgcc gatgattaat tgtcaacacg tgctcgagct
     2341 agctggcggg aacggcaatg ctcctggcgg cggcggggct ggtgcccagg tctccgcaca
     2401 gagcggcggt gctggcgctc gcggccaggc gtggttcttc gcgtactgac aagaaacccc
     2461 cctctttagg actcagtgtc cttgggaggg gggctttttg cgtttcagga ggtcttggcc
     2521 agcttggaca tcgcctcagc gatagcctcg tcgcgggcct cagacgccat ctggtacttc
     2581 atcgccatcc taggagtcgt gtgaccgaga cgggccatca gctccttggt cgtcgcacct
     2641 gcctgagcgg cgaacgtagc gccgacagcg cggaggtcgt ggatgcggag ttccggccga
     2701 ccgatcttgg cgtagccacg cttcagcgac ttggtgaacg cggacttcga cagccggttg
     2761 ccctgcgtcg tggtcaccag gaatgcctcg gggcccttgt tcatcttcgt acggtccttc
     2821 atgtgcgctc ggatcatctc cgcgacgtga ggcggaaccg tcacaggacg cttcgaccgg
     2881 acggtcttgg cgttgccaac gacgatcttg ttccccacgc gggaagcgcc acggcgcacc
     2941 cggagcttca tcgtcatgcc gtcgtccacg atgtccttgc ggcgaagctc gatcagctct
     3001 ccgaaccgga ggctcgtcca cgccaggatg tatgccgcga tccggtagtg cacgaagatc
     3061 tcagcggcga cgatgtccag ctcctcaggc gtcagcgcct ctacgtcgcg ctcatcggct
     3121 gccttctgct cgatccggca cgggttctct gcgatcagct tgtcctcgac cgctgtgttc
     3181 atcaccgccc ggaggacgtt gtaggcatgc cggcgggcag tcgggtgctt cctacccatc
     3241 ccggcccacc acgcacgcac cagagctggc gtcatctctg tgaccgccac ttcacctagc
     3301 accgggtaga tgcggcgctc cgcgtgcccg ctgtacagat ccctggtgcc gtctgcgagg
     3361 tcgcgctcca cgagccactt ccgggtgtac tcctccagcg tgatggcgct ggcggctgcc
     3421 ttcttcgccc ggtcctgtgg aggggtccag gtctccatct cgatgagccg cttctcgccc
     3481 gcgagccagg cttcggcgtc catcttgttg tcgtaggtct gcagcgcgta gtacctcaca
     3541 ccgtcctgcg ggttgacgta tgaggcttgg atcctcccgc tgcgctgagt cttcagcgat
     3601 ccccatccgc gacgtgccaa ctaggtctcc tctcgtcgtg aacaaggcta ccgggttgca
     3661 actcctgtgc aactctcagg cttcaacgcg cttctacgac ctgcaatttc tttccactta
     3721 gaggatgcag ccgagagggg gtaaaaacct atcttgaccg gcccatatat gtggtcggca
     3781 gacacccatt cttccaaact agctacgcgg gttcgattcc cgtcgcccgc tccgctggtc
     3841 agagggtgtt ttcgccctct ggccattttt ctttccaggg gtctgcaact cttgtgcgac
     3901 tcttctgacc tgggcatacg cggttgcaac gcatccctga tctggctact ttcgatgctg
     3961 acaaacgaat agagcccccc gcctgcgcga acagacgagg ggcattcaca ccagattgga
     4021 gctggtgcag tgaagagaat agaccgggac aaggttgcac cgggagttgc agcggtcgga
     4081 accctcgccg tcggcgggct ggcgttcgcc ctgtcgttca cggctctagc tgatcaccgc
     4141 ggccatgatg gcataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
     4201 tgcctggcag tcgatcgttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     4261 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     4321 ggcgccgggg gcggtgtccg gcggccccca gaggaactgc gccagttcct ccggatcggt
     4381 gaagccggag agatccagcg gggtctcctc gaacacctcg aagtcgtgca ggaaggtgaa
     4441 ggcgagcagt tcgcgggcga agtcctcggt ccgcttccac tgcgccccgt cgagcagcgc
     4501 ggccaggatc tcgcggtcgc cccggaaggc gttgagatgc agttgcacca ggctgtagcg
     4561 ggagtctccc gcatagacgt cggtgaagtc gacgatcccg gtgacctcgg tcgcggccag
     4621 gtccacgaag gtgttggtcc cgtgcaggtc gccgtggacg aaccggggtt cgcggccggc
     4681 cagcagcgtg tccacgtccg gcagccagtc ctccaggcgg tccagcagcc ggggcgagag
     4741 gtagccctac ccgcggtggt cctcgacggt cgccgcgcgg cgttcccgca gcagttccgg
     4801 gaagacctcg gaatggg
//
