LOCUS       sequence-291818-        3158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orbit integrating plasmid for C-terminal tagging with degradation 
            tag DAS+4, using hygromycin as a selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3158)
  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 3158)
  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: pKM446 (Flag-DAS+4))
FEATURES             Location/Qualifiers
     source          1..3158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    47..84
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             87..110
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      complement(381..969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(461..480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1234..1253)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3106..3152)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagtcgat cgtacgctag ttaactacgt cgacatcgat aagcttggct tgtcgacgac
       61 ggcggtctcc gtcgtcagga tcatcggact acaaggacga cgacgacaag gccgccaacg
      121 acgagaacta ctccgagaac tacgcggacg ccagctagtg actcgagtct agagcatgca
      181 ctagtgcggc cgctagcggt accagatctg ggcccttaag tcgcgttgcg ctcggtcgtt
      241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      541 gcctttctcc cctcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      781 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
      841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcatgg aactagagca
     1021 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatcg gcacgtaaga
     1081 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     1141 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1201 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1261 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1321 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1381 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1441 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1501 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1561 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1621 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1681 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1741 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1801 tgcgatgagt ggctggcgga aacggcaatg ctgaagtttt aaatcaatct aaagtatata
     1861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1921 ctgtctattt cgttcatcca tagttgcctg actccccgac gtggccgacc agcccgtcat
     1981 cgtcaacgcc tgatccgcgg tgcggacagg ccgtgtcgcg accggccgtg cggaattaag
     2041 ccggcccgta ccctgtgaat agaggtccgc tgtgacacaa gaatccctgt tacttctcga
     2101 ccgtattgat tcggatgatt cctacgcgag cctgcggaac gaccaggaat tctgggagcc
     2161 gctggcccgc cgagccctgg aggagctcgg gctgccggtg ccgccggtgc tgcgggtgcc
     2221 cggcgagagc accaaccccg tactggtcgg cgagcccggc ccggtgatca agctgttcgg
     2281 cgagcactgg tgcggtccgg agagcctcgc gtcggagtcg gaggcgtacg cggtcctggc
     2341 ggacgccccg gtgccggtgc cccgcctcct cggccgcggc gagctgcggc ccggcaccgg
     2401 agcctggccg tggccctacc tggtgatgag ccggatgacc ggcaccacct ggcggtccgc
     2461 gatggacggc acgaccgacc ggaacgcgct gctcgccctg gcccgcgaac tcggccgggt
     2521 gctcggccgg ctgcacaggg tgccgctgac cgggaacacc gtgctcaccc cccattccga
     2581 ggtcttcccg gaactgctgc gggaacgccg cgcggcgacc gtcgaggacc accgcgggtg
     2641 gggctacctc tcgccccggc tgctggaccg cctggaggac tggctgccgg acgtggacac
     2701 gctgctggcc ggccgcgaac cccggttcgt ccacggcgac ctgcacggga ccaacatctt
     2761 cgtggacctg gccgcgaccg aggtcaccgg gatcgtcgac ttcaccgacg tctatgcggg
     2821 agactcccgc tacagcctgg tgcaactgca tctcaacgcc ttccggggcg accgcgagat
     2881 cctggccgcg ctgctcgacg gggcgcagtg gaagcggacc gaggacttcg cccgcgaact
     2941 gctcgccttc accttcctgc acgacttcga ggtgttcgag gagaccccgc tggatctctc
     3001 cggcttcacc gatccggagg aactggcgca gttcctctgg gggccgccgg acaccgcccc
     3061 cggcgcctga cgccccctct agctgatcac cgcggccatg atggcataaa acgaaaggcc
     3121 cagtctttcg actgagcctt tcgttttatt tgatgcct
//
