LOCUS       sequence-210531-        3571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORBIT integrating plasmid for C-terminal tagging with Myc-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3571)
  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 3571)
  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: pKM492-Myc-His
FEATURES             Location/Qualifiers
     source          1..3571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    440..477
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             510..539
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             540..557
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      600..627
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(794..1382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(874..893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1647..1666)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3519..3565)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagtcgat cgagaggtga ccacaacgac gcgcccgctt tgatcgggga cgtctgcggc
       61 cgaccattta cgggtcttgt tgtcgttggc ggtcatgggc cgaacatact cacccggatc
      121 ggagggccga ggacaaggtc gaacgagggg catgacccgg tgcggggctt cttgcactcg
      181 gcataggcga gtgctaagaa taacgttggc actcgcgacc ggtgagtgct aggtcgggac
      241 ggtgaggcca ggcccgtcgt cgcagcgagt ggcagcgagg acaacttgag ccgtccgtcg
      301 cgggcactgc gcccggccag cgtaagtagc ggggttgccg tcacccggtg acccccgttt
      361 catccccgat ccggaggaat cacttcgcaa tggccaagac aattgcggat ccatcgatag
      421 aaaggaggaa ggaaagcttg gcttgtcgac gacggcggtc tccgtcgtca ggatcatcgg
      481 gcggcggcgg cgagaacctg tacttccagg agcagaagct gatcagcgag gaagacctgc
      541 accaccacca ccaccactga tgactcgagt ctagagcatg cactagtgcg gccgctagca
      601 gaaggccatc ctgacggatg gccttttctt aagtcgcgtt gcgctcggtc gttcggctgc
      661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      961 tcccctcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1201 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     1261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1381 aagaagatcc tttgatcttt tctacggggt ctgacgctca tggaactaga gcatgagatt
     1441 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tcggcacgta agaggttcca
     1501 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     1561 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     1621 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     1681 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     1741 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     1801 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     1861 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     1921 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     1981 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     2041 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     2101 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     2161 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     2221 agtggctggc ggaaacggca atgctgaagt tttaaatcaa tctaaagtat atatgagtaa
     2281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2341 tttcgttcat ccatagttgc ctgactcccc gacgtggccg accagcccgt catcgtcaac
     2401 gcctgatccg cggtgcggac aggccgtgtc gcgaccggcc gtgcggaatt aagccggccc
     2461 gtaccctgtg aatagaggtc cgctgtgaca caagaatccc tgttacttct cgaccgtatt
     2521 gattcggatg attcctacgc gagcctgcgg aacgaccagg aattctggga gccgctggcc
     2581 cgccgagccc tggaggagct cgggctgccg gtgccgccgg tgctgcgggt gcccggcgag
     2641 agcaccaacc ccgtactggt cggcgagccc ggcccggtga tcaagctgtt cggcgagcac
     2701 tggtgcggtc cggagagcct cgcgtcggag tcggaggcgt acgcggtcct ggcggacgcc
     2761 ccggtgccgg tgccccgcct cctcggccgc ggcgagctgc ggcccggcac cggagcctgg
     2821 ccgtggccct acctggtgat gagccggatg accggcacca cctggcggtc cgcgatggac
     2881 ggcacgaccg accggaacgc gctgctcgcc ctggcccgcg aactcggccg ggtgctcggc
     2941 cggctgcaca gggtgccgct gaccgggaac accgtgctca ccccccattc cgaggtcttc
     3001 ccggaactgc tgcgggaacg ccgcgcggcg accgtcgagg accaccgcgg gtggggctac
     3061 ctctcgcccc ggctgctgga ccgcctggag gactggctgc cggacgtgga cacgctgctg
     3121 gccggccgcg aaccccggtt cgtccacggc gacctgcacg ggaccaacat cttcgtggac
     3181 ctggccgcga ccgaggtcac cgggatcgtc gacttcaccg acgtctatgc gggagactcc
     3241 cgctacagcc tggtgcaact gcatctcaac gccttccggg gcgaccgcga gatcctggcc
     3301 gcgctgctcg acggggcgca gtggaagcgg accgaggact tcgcccgcga actgctcgcc
     3361 ttcaccttcc tgcacgactt cgaggtgttc gaggagaccc cgctggatct ctccggcttc
     3421 accgatccgg aggaactggc gcagttcctc tgggggccgc cggacaccgc ccccggcgcc
     3481 tgacgccccc tctagctgat caccgcggcc atgatggcat aaaacgaaag gcccagtctt
     3541 tcgactgagc ctttcgtttt atttgatgcc t
//
