LOCUS       sequence-210525-        3062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORBIT integrating plasmid for deletion of target gene, using zeocin 
            as a selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3062)
  AUTHORS   Murphy KC, Nelson SJ, Nambi S, Papavinasasundaram K, Baer CE, 
            Sassetti CM
  TITLE     ORBIT: a New Paradigm for Genetic Engineering of Mycobacterial 
            Chromosomes.
  JOURNAL   MBio. 2018 Dec 11;9(6). pii: mBio.01467-18. doi: 
            10.1128/mBio.01467-18.
  PUBMED    30538179
REFERENCE   2  (bases 1 to 3062)
  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: "MBio."; 
            date: "2018-12-11"; volume: "9(6). pii: mBio.01467-18. doi"; pages: 
            " 10.1128/mBio.01467-18"
COMMENT     Sequence Label: pKM496 (deletions) Zeo
FEATURES             Location/Qualifiers
     source          1..3062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(80..126)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    572..609
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      646..673
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(840..1428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(920..939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1693..1712)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        2461..2508
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2527..2901
                     /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"
     terminator      2943..3029
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgcagttcct ctgggggccg ccggacaccg cccccggcgc ctgacgcccc ctctagctga
       61 tcaccgcggc catgatggca taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
      121 tatttgatgc ctggcagtcg atcgagaggt gaccacaacg acgcgcccgc tttgatcggg
      181 gacgtctgcg gccgaccatt tacgggtctt gttgtcgttg gcggtcatgg gccgaacata
      241 ctcacccgga tcggagggcc gaggacaagg tcgaacgagg ggcatgaccc ggtgcggggc
      301 ttcttgcact cggcataggc gagtgctaag aataacgttg gcactcgcga ccggtgagtg
      361 ctaggtcggg acggtgaggc caggcccgtc gtcgcagcga gtggcagcga ggacaacttg
      421 agccgtccgt cgcgggcact gcgcccggcc agcgtaagta gcggggttgc cgtcacccgg
      481 tgacccccgt ttcatccccg atccggagga atcacttcgc aatggccaag acaattgcgg
      541 atccatcgat agaaaggagg aaggaaagct tggcttgtcg acgacggcgg tctccgtcgt
      601 caggatcatc tcgagtctag agcatgcact agtgcggccg ctagcagaag gccatcctga
      661 cggatggcct tttcttaagt cgcgttgcgc tcggtcgttc ggctgcggcg agcggtatca
      721 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      781 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      841 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      901 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      961 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ctcgggaagc
     1021 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1081 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1141 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1201 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1261 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1321 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1381 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1441 atcttttcta cggggtctga cgctcatgga actagagcat gagattatca aaaaggatct
     1501 tcacctagat ccttttaaat taaaaatcgg cacgtaagag gttccaactt tcaccataat
     1561 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     1621 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1681 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1741 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1801 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     1861 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1921 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1981 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     2041 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     2101 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     2161 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     2221 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gctggcggaa
     2281 acggcaatgc tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2401 agttgcctga ctccccgacg tggccgacca gcccgtcatc gtcaacgcct gaccgcggtt
     2461 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     2521 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     2581 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     2641 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     2701 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     2761 tcggaggtcg tgtccacgaa cttccgggac gcctctgggc cggccatgac cgagatcggc
     2821 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     2881 gtggccgagg agcaggactg aatgcatctc tccccatgcg agagtaggga actgccaggc
     2941 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3001 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaact
     3061 gg
//
