LOCUS       sequence-210545-        3635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORBIT integrating plasmid for C-terminal tagging with SNAP tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3635)
  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 3635)
  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: pKM489-Snap
FEATURES             Location/Qualifiers
     source          1..3635
                     /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..632
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     rep_origin      complement(858..1446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(938..957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1711..1730)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3583..3629)
                     /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 tcatcgatgg acaaagattg cgaaatgaaa cgtaccaccc
      121 tggatagccc gctgggcaaa ctggaactga gcggctgcga acagggcctg catgaaatta
      181 aactgctggg taaaggcacc agcgcggccg atgcggttga agttccggcc ccggccgccg
      241 tgctgggtgg tccggaaccg ctgatgcagg cgaccgcgtg gctgaacgcg tattttcatc
      301 agccggaagc gattgaagaa tttccggttc cggcgctgca tcatccggtg tttcagcagg
      361 agagctttac ccgtcaggtg ctgtggaaac tgctgaaagt ggttaaattt ggcgaagtga
      421 ttagctatca gcagctggcg gccctggcgg gtaatccggc ggccaccgcc gccgttaaaa
      481 ccgcgctgag cggtaacccg gtgccgattc tgattccgtg ccatcgtgtg gttagctcta
      541 gcggtgcggt tggcggttat gaaggtggtc tggcggtgaa agagtggctg ctggcccatg
      601 aaggtcatcg tctgggtaaa ccgggtctgg gatagtgact cgagtctaga gcatgcacta
      661 gtgcggccgc tagcggtacc agatctgggc ccttaagtcg cgttgcgctc ggtcgttcgg
      721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1021 tttctcccct cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1261 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     1321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1381 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1441 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcatggaac tagagcatga
     1501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatcggca cgtaagaggt
     1561 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     1621 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     1681 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     1741 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     1801 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     1861 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     1921 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     1981 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     2041 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     2101 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     2161 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     2221 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     2281 gatgagtggc tggcggaaac ggcaatgctg aagttttaaa tcaatctaaa gtatatatga
     2341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2401 tctatttcgt tcatccatag ttgcctgact ccccgacgtg gccgaccagc ccgtcatcgt
     2461 caacgcctga tccgcggtgc ggacaggccg tgtcgcgacc ggccgtgcgg aattaagccg
     2521 gcccgtaccc tgtgaataga ggtccgctgt gacacaagaa tccctgttac ttctcgaccg
     2581 tattgattcg gatgattcct acgcgagcct gcggaacgac caggaattct gggagccgct
     2641 ggcccgccga gccctggagg agctcgggct gccggtgccg ccggtgctgc gggtgcccgg
     2701 cgagagcacc aaccccgtac tggtcggcga gcccggcccg gtgatcaagc tgttcggcga
     2761 gcactggtgc ggtccggaga gcctcgcgtc ggagtcggag gcgtacgcgg tcctggcgga
     2821 cgccccggtg ccggtgcccc gcctcctcgg ccgcggcgag ctgcggcccg gcaccggagc
     2881 ctggccgtgg ccctacctgg tgatgagccg gatgaccggc accacctggc ggtccgcgat
     2941 ggacggcacg accgaccgga acgcgctgct cgccctggcc cgcgaactcg gccgggtgct
     3001 cggccggctg cacagggtgc cgctgaccgg gaacaccgtg ctcacccccc attccgaggt
     3061 cttcccggaa ctgctgcggg aacgccgcgc ggcgaccgtc gaggaccacc gcgggtgggg
     3121 ctacctctcg ccccggctgc tggaccgcct ggaggactgg ctgccggacg tggacacgct
     3181 gctggccggc cgcgaacccc ggttcgtcca cggcgacctg cacgggacca acatcttcgt
     3241 ggacctggcc gcgaccgagg tcaccgggat cgtcgacttc accgacgtct atgcgggaga
     3301 ctcccgctac agcctggtgc aactgcatct caacgccttc cggggcgacc gcgagatcct
     3361 ggccgcgctg ctcgacgggg cgcagtggaa gcggaccgag gacttcgccc gcgaactgct
     3421 cgccttcacc ttcctgcacg acttcgaggt gttcgaggag accccgctgg atctctccgg
     3481 cttcaccgat ccggaggaac tggcgcagtt cctctggggg ccgccggaca ccgcccccgg
     3541 cgcctgacgc cccctctagc tgatcaccgc ggccatgatg gcataaaacg aaaggcccag
     3601 tctttcgact gagcctttcg ttttatttga tgcct
//
