LOCUS       sequence-210529-        3635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORBIT integrating plasmid for C-terminal tagging with CLIP 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: pKM490-Clip
FEATURES             Location/Qualifiers
     source          1..3635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    41..78
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             81..626
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     rep_origin      complement(852..1440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(932..951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1705..1724)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3577..3623)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgatcgtacg ctagttaact acgtcgacat cgataagctt ggcttgtcga cgacggcggt
       61 ctccgtcgtc aggatcatcg atggacaaag actgcgaaat gaagcgcacc accctggata
      121 gccctctggg caagctggaa ctgtctgggt gcgaacaggg cctgcaccgt atcatcttcc
      181 tgggcaaagg aacatctgcc gccgacgccg tggaagtgcc tgccccagcc gccgtgctgg
      241 gcggaccaga gccactgatc caggccaccg cctggctcaa cgcctacttt caccagcctg
      301 aggccatcga ggagttccct gtgccagccc tgcaccaccc agtgttccag caggagagct
      361 ttacccgcca ggtgctgtgg aaactgctga aagtggtgaa gttcggagag gtcatcagcg
      421 agagccacct ggccgccctg gtgggcaatc ccgccgccac cgccgccgtg aacaccgccc
      481 tggacggaaa tcccgtgccc attctgatcc cctgccaccg ggtggtgcag ggcgacagcg
      541 acgtggggcc ctacctgggc gggctcgccg tgaaagagtg gctgctggcc cacgagggcc
      601 acagactggg caagcctggg ctgggttagt gactcgagtc tagagcatgc actagtgcgg
      661 ccgctagcgg taccagatct gggcccttaa gtcgcgttgc gctcggtcgt tcggctgcgg
      721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1021 ccctcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1261 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     1321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1441 gaagatcctt tgatcttttc tacggggtct gacgctcatg gaactagagc atgagattat
     1501 caaaaaggat cttcacctag atccttttaa attaaaaatc ggcacgtaag aggttccaac
     1561 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     1621 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     1681 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     1741 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     1801 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     1861 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     1921 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     1981 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     2041 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     2101 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     2161 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     2221 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     2281 tggctggcgg aaacggcaat gctgaagttt taaatcaatc taaagtatat atgagtaaac
     2341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2401 tcgttcatcc atagttgcct gactccccga cgtggccgac cagcccgtca tcgtcaacgc
     2461 ctgatccgcg gtgcggacag gccgtgtcgc gaccggccgt gcggaattaa gccggcccgt
     2521 accctgtgaa tagaggtccg ctgtgacaca agaatccctg ttacttctcg accgtattga
     2581 ttcggatgat tcctacgcga gcctgcggaa cgaccaggaa ttctgggagc cgctggcccg
     2641 ccgagccctg gaggagctcg ggctgccggt gccgccggtg ctgcgggtgc ccggcgagag
     2701 caccaacccc gtactggtcg gcgagcccgg cccggtgatc aagctgttcg gcgagcactg
     2761 gtgcggtccg gagagcctcg cgtcggagtc ggaggcgtac gcggtcctgg cggacgcccc
     2821 ggtgccggtg ccccgcctcc tcggccgcgg cgagctgcgg cccggcaccg gagcctggcc
     2881 gtggccctac ctggtgatga gccggatgac cggcaccacc tggcggtccg cgatggacgg
     2941 cacgaccgac cggaacgcgc tgctcgccct ggcccgcgaa ctcggccggg tgctcggccg
     3001 gctgcacagg gtgccgctga ccgggaacac cgtgctcacc ccccattccg aggtcttccc
     3061 ggaactgctg cgggaacgcc gcgcggcgac cgtcgaggac caccgcgggt ggggctacct
     3121 ctcgccccgg ctgctggacc gcctggagga ctggctgccg gacgtggaca cgctgctggc
     3181 cggccgcgaa ccccggttcg tccacggcga cctgcacggg accaacatct tcgtggacct
     3241 ggccgcgacc gaggtcaccg ggatcgtcga cttcaccgac gtctatgcgg gagactcccg
     3301 ctacagcctg gtgcaactgc atctcaacgc cttccggggc gaccgcgaga tcctggccgc
     3361 gctgctcgac ggggcgcagt ggaagcggac cgaggacttc gcccgcgaac tgctcgcctt
     3421 caccttcctg cacgacttcg aggtgttcga ggagaccccg ctggatctct ccggcttcac
     3481 cgatccggag gaactggcgc agttcctctg ggggccgccg gacaccgccc ccggcgcctg
     3541 acgccccctc tagctgatca ccgcggccat gatggcataa aacgaaaggc ccagtctttc
     3601 gactgagcct ttcgttttat ttgatgcctg gcagt
//
